CN105963325A

CN105963325A - Application of sulfated polysaccharides from seaweed

Info

Publication number: CN105963325A
Application number: CN201610298717.8A
Authority: CN
Inventors: 王长振; 王水明; 郑文; 胡向军; 周红梅; 王丽峰; 李杨; 徐新萍; 邹勇; 苏慧婷; 智维佳; 张潇
Original assignee: Institute of Radiation Medicine of CAMMS
Current assignee: Institute of Radiation Medicine of CAMMS
Priority date: 2016-05-06
Filing date: 2016-05-06
Publication date: 2016-09-28

Abstract

The invention relates to the field of medicament technology, and concretely relates to an application of sulfated polysaccharides from seaweed. The sulfated polysaccharides from seaweed are applied to prevention and treatment of microwave radiation. A series of experiment researches prove that sulfated polysaccharides from seaweed have good effects in the aspect of prevention and treatment of microwave radiation.

Description

A kind of application of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS

Technical field

The present invention relates to technical field of pharmaceuticals, be specifically related to the application of a kind of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS.

Background technology

Massive epidemiology is investigated, be experimental studies have found that, electromagnetic radiation can damage multiple organ system, such as nervous system, life Grow system, immune system etc., cause DNA damage and protein unconventionality expression etc..2011, IRAC (International Agency for Research on Cancer, IARC) electromagnetic radiation is classified as it is potentially carcinogenic thing, more Cause people's common concern to electromagnetic radiation.Currently, electromagnetic radiation damage mechanism illustrates the most completely.Research is had to point out, oxygen Change one of main mechanism of stress damage HPM irradiating biological damage effect.

High power microwave weapon is one of Developing the most rapid new and high technology weapon, is modern and Future Informationization war The significant weapon striven, not only disturbs and destroys armament systems electronic equipment/equipment, also can cause flat, army personnel in wartime Damage.Existing result of study shows, HPM radiation can damage multiple organ system, such as nervous system, reproductive system, immune system Deng, but its damage mechanisms illustrates the most completely, oxidativestress damage, apoptosis-related protein caused by energy metabolism impairment, free radical Unconventionality expression, DNA damage, signal transduction pathway exception etc. plays a significant role wherein, and oxidativestress damage is HPM radiation One of main mechanism of biological damage effect.The preventing and treating of HPM radiation damage in addition to shielding is with the physical protection such as absorption, current city Evident in efficacy and that safety is high protective agents is there is no on face.

SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS, complete entitled Sargassum polysaccharides sulfuric ester, is a kind of macromolecular polysaccharide complex extracted from Thallus Laminariae (Thallus Eckloniae). Early-stage Study shows, SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS energy scavenging hydroxyl and ultra-oxygen anion free radical, reduces malonaldehyde (MDA) and egg White matter carbonyl (PCO) content, raises the activities of antioxidant enzymes such as glutathione peroxidase (GPX), catalase (CAT), And have and have no side effect, crude drug is cheap, it is simple to preparation, it is easy to the many merits such as use.Real according to early stage exploration Sensitive indicator determined by testing and sensitive internal organs, this part experiment is drenched by amynologic index routine blood test, CD3+, CD4+, CD8+ Bar cell subsets and oxidative stress index XOD, MPO, SOD, GSH-PX, CAT, MDA, PCO content or Activity determination, in conjunction with Hippocampus With the morphological change of testis tissue, determine SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS to brain and testis oxidativestress damage effective dose, for finally Obtain army spy medicine clinic official written reply to lay a solid foundation.

Summary of the invention

It is an object of the invention to provide the application of a kind of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS, it was demonstrated that SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS is at preventing and treating microwave spoke Penetrate aspect and there is good effect.

In order to realize object above, the present invention adopts the following technical scheme that

A kind of application of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS, the application in terms of preventing and treating microwave radiation of the described SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS.

Optionally, described sulphuric acid Sargassum polysaccharides application on the medicine of preparation preventing and treating microwave radiation.

Optionally, in described microwave radiation microwave be frequency 2-4GHz, wavelength be the S-band microwave of 150-75.00mm.

Optionally, described microwave is the microwave of frequency 2.856GHz.

Optionally, described microwave average power density is 30mW/cm², peak power density is 200mW/cm², pulse frequency For 300pps, pulsewidth is 500ns.

Sargassum polysaccharides the most used is by following extracting method extraction gained:

(1) after Thallus Laminariae (Thallus Eckloniae) being placed in the water of himself quality 38-42 times immersion 35-45 minute, for himself matter The water of amount 38-42 times amount is rinsed well；

(2) Thallus Laminariae (Thallus Eckloniae) is carried out pretreatment, reject the defective Thallus Laminariae (Thallus Eckloniae) that leaf is yellow, leaf is broken and mouldy, qualified Thallus Laminariae (Thallus Eckloniae) is cut into face The fragment of long-pending 7-10 square centimeter；

(3) Thallus Laminariae (Thallus Eckloniae) is joined in the water of himself quality 9-11 times decoct three times, respectively 1.9-2.1 hour, 1.4-1.6 hour, 1.4-1.6 hour；

(4) it is evaporated to relative density 1.05-1.15 after three decoction liquor being merged and obtains concentrated solution I；

(5) in concentrated solution I, add ethanol to ethanol volumetric concentration 29-31%, after standing overnight centrifugal supernatant and Sediment；

(6) sediment is detected, if SO in sediment₄ ^2-Weight/mass percentage composition more than 5%, in sediment, add ethanol To ethanol volumetric concentration 29-31%, stand to layering and be centrifuged to obtain supernatant；If SO in sediment₄ ^2-Weight/mass percentage composition little In equal to 5%, abandon sediment；

(7) being merged by the supernatant in step S50 and step S60, being evaporated to relative density 1.1-1.2 must concentrate Liquid II；

(8) in concentrated solution II, addition ethanol, to ethanol volumetric concentration 83-86%, after standing overnight, abandons supernatant, heavy Form sediment with centrifugal after the washing with alcohol of percentage by volume 74-76%；

(9) carry out being spray-dried to obtain SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS after the water dissolution of the precipitation 7-9 times amount after being centrifuged.

In said extracted method, after Thallus Laminariae (Thallus Eckloniae) being placed in the water of himself quality 40 times by step (1) immersion 40 minutes, Water for himself quality 40 times amount is rinsed well；

In step (2), qualified Thallus Laminariae (Thallus Eckloniae) is cut into the fragment of area 9 square centimeters；

In step (3), Thallus Laminariae (Thallus Eckloniae) is joined in the water of himself quality 10, decocts three times, respectively 2 hours, 1.5 little Time, 1.5 hours；

In step (4), it is evaporated to relative density 1.1 after three decoction liquor being merged and obtains concentrated solution I；

In step (5), in concentrated solution I, addition ethanol is to ethanol volumetric concentration 30%, is centrifuged and obtains supernatant after standing overnight Liquid and sediment；

In step (6), if SO in sediment₄ ^2-Weight/mass percentage composition more than 5%, in sediment, add ethanol to ethanol body Volume concentrations 30%, stands to layering and is centrifuged to obtain supernatant；

In step (7), the supernatant in step S50 and step S60 is merged, be evaporated to relative density 1.17 and obtain dense Contracting liquid II；

In step (8), in concentrated solution II, addition ethanol is to ethanol volumetric concentration 85%, after standing overnight, abandons supernatant, Precipitation is with centrifugal after the washing with alcohol of percentage by volume 75%；

In step (9), will centrifugal after precipitation be spray-dried to obtain SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS with carrying out after the water dissolution of 8 times amount.

The present invention is proved by series of experiment research, and SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS has well in terms of preventing and treating microwave radiation Effect, specify that the SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS effective dose to HPM radiation damage preventive and therapeutic effect, it was demonstrated that middle dosage (50mg/kg/d) Medicine group is its optimal administration concentration.Meanwhile, SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS can reduce the free radicals such as XOD, MPO and generate related enzyme activity, Increase the activities of antioxidant enzymes such as SOD, CAT, reduce the Radical Metabolism product assay such as MDA, PCO, body caused by its preventing and treating HPM One of mechanism of oxidative damage.

Accompanying drawing explanation

Body weight change during the administration of Fig. 1 rat oral gavage；

1d after Fig. 2 Normal group drug withdrawal, the most structure of hippocampal neuron is normal, and perivascular space is the most broadening；

3d after Fig. 3 Normal group drug withdrawal, hippocampal neural meta structure is normal, and perivascular space is the most broadening；

1d after Fig. 4 radiation group drug withdrawal, hippocampal neuron pyknosis deeply contaminates and increases, and cell and perivascular space are the most broadening；

3d after Fig. 5 radiation group drug withdrawal, hippocampal neuron pyknosis deeply contaminates showed increased, cell and perivascular space and substantially increases Wide；

7d after Fig. 6 radiation group drug withdrawal, it is the most broadening that hippocampal neuron pyknosis contaminates minimizing, cell and perivascular space deeply；

1d after Fig. 7 low-dose drugs group drug withdrawal, hippocampal neuron pyknosis deeply contaminates and increases, and cell and perivascular space slightly increase Wide；

3d after Fig. 8 low-dose drugs group drug withdrawal, hippocampal neuron pyknosis deeply contaminates and increases, and cell and perivascular space slightly increase Wide；

7d after Fig. 9 low-dose drugs group drug withdrawal, hippocampal neuron pyknosis significantly reduces, and cell and perivascular space slightly increase Wide；

1d after the drug withdrawal of dose drug group in Figure 10, minority hippocampal neuron pyknosis contaminates deeply, and perivascular space is the most broadening；

3d after the drug withdrawal of dose drug group in Figure 11, minority hippocampal neuron pyknosis contaminates deeply, and perivascular space is the most broadening；

7d after the drug withdrawal of dose drug group in Figure 12, hippocampal neural meta structure is normal, and perivascular space is the most broadening；

1d after Figure 13 high dose medicament group drug withdrawal, minority hippocampal neuron pyknosis contaminates deeply, and perivascular space is the most broadening；

3d after Figure 14 high dose medicament group drug withdrawal, part hippocampal neuron pyknosis contaminates deeply, and perivascular space is the most broadening；

7d after Figure 15 high dose medicament group drug withdrawal, hippocampal neural meta structure is normal, and perivascular space is the most broadening；

1d after Figure 16 positive drug group drug withdrawal, minority hippocampal neuron pyknosis contaminates deeply, and cell and perivascular space slightly increase Wide；

1d after Figure 17 positive drug group drug withdrawal, minority hippocampal neuron pyknosis contaminates deeply, and cell and perivascular space slightly increase Wide；

1d after Figure 18 positive drug group drug withdrawal, minority hippocampal neuron pyknosis contaminates deeply, and cell and perivascular space slightly increase Wide；

1d after the drug withdrawal of dose drug matched group in Figure 19, hippocampal neural meta structure is normal, and perivascular space slightly increases Wide；

7d after the drug withdrawal of dose drug matched group in Figure 20, hippocampal neural meta structure is normal, and perivascular space slightly increases Wide；

1d after Figure 21 Normal group drug withdrawal, testicular spermatogenic tubule spermatogenic cells at different stages structure is normal, clear layer, sperm Abundant；

1d after Figure 22 radiation group drug withdrawal, the obvious edema of spermatogenic epithelium layer, spermatogenic cell arrangement is loose, spermatid and sperm Reduce in various degree；

3d after Figure 23 radiation group drug withdrawal, spermatogenic epithelium layer edema in seminiferous tubule, albumen edematous fluid accumulation seen from intracavity；

7d after Figure 24 radiation group drug withdrawal, in seminiferous tubule, spermatogenic epithelium layer edema and the accumulation of albumen edematous fluid still suffer from, but Alleviate；

1d after Figure 25 low-dose drugs group drug withdrawal, spermatogenic epithelium layer edema, albumen edematous fluid accumulation seen from intracavity；

3d after Figure 26 low-dose drugs group drug withdrawal, the obvious edema of spermatogenic epithelium layer, spermatogenic cell arrangement is loose；

7d after Figure 27 low-dose drugs group drug withdrawal, the obvious edema of spermatogenic epithelium layer, spermatogenic cell arrangement is loose；

1d after the drug withdrawal of dose drug group, the light intermediate edema of spermatogenic epithelium layer in Figure 28；

3d after the drug withdrawal of dose drug group, spermatogenic epithelium layer Mild edema in Figure 29；

7d after the drug withdrawal of dose drug group in Figure 30, seminiferous tubule spermatogenic cells at different stages structure is normal；

1d after Figure 31 high dose medicament group drug withdrawal, the more apparent edema of spermatogenic epithelium layer；

3d after Figure 32 high dose medicament group drug withdrawal, the light intermediate edema of spermatogenic epithelium layer；

7d after the drug withdrawal of dose drug group in Figure 33, spermatogenic epithelium layer edema recovers normal the most completely；

1d after Figure 34 positive drug group drug withdrawal, the more apparent edema of spermatogenic epithelium layer；

3d after Figure 35 positive drug group drug withdrawal, the more apparent edema of spermatogenic epithelium layer, albumen edematous fluid gathers；

7d after Figure 36 positive drug group drug withdrawal, spermatogenic epithelium layer edema recovers normal the most completely；

1d after the drug withdrawal of dose drug matched group in Figure 37, testicular spermatogenic tubule spermatogenic cells at different stages structure is normal；

7d after the drug withdrawal of dose drug matched group in Figure 38, testicular spermatogenic tubule spermatogenic cells at different stages structure is normal；

1d after Figure 39 Normal group drug withdrawal, granular cell morphosis is normal；

1d after Figure 40 Normal group drug withdrawal, pyramidal cell morphosis is normal, mitochondrion mild swelling；

7d after Figure 41 Normal group drug withdrawal, glial cell morphosis is normal；

1d after Figure 42 Normal group drug withdrawal, perivascular space is normal；

1d after Figure 43 Normal group drug withdrawal, synaptic structure is the fuzzyyest；

7d after Figure 44 Normal group drug withdrawal, glial cell Mild edema；

1d after Figure 45 radiation group drug withdrawal, hippocampal neuron nuclear chromatin loosens, mitochondrial swelling, cavitation, ridge disorder, lyase Body increases；

7d after Figure 46 radiation group drug withdrawal, glial cell obvious tumefaction, endochylema organelle is sparse；

1d after Figure 47 radiation group drug withdrawal, synaptic space obscures；

1d after Figure 48 radiation group drug withdrawal, perivascular space is the most broadening；

1d after Figure 49 low-dose drugs group drug withdrawal, granular cell mitochondrial swelling, endoplasmic reticulum and Golgi body are slightly expanded；

1d after Figure 50 low-dose drugs group drug withdrawal, granular cell obvious tumefaction, takes off change；

1d after Figure 51 low-dose drugs group drug withdrawal, granular cell chromatic agglutination limit is moved, in apoptosis；

1d after Figure 52 low-dose drugs group drug withdrawal, pyramidal cell mitochondrion obvious tumefaction, neuronophagia occurs；

1d after Figure 53 low-dose drugs group drug withdrawal, synaptic space obscures；

1d after Figure 54 radiation group drug withdrawal, perivascular space is the most broadening；

1d after the drug withdrawal of dose drug group, granular cell mitochondrion mild swelling in Figure 55；

7d after the drug withdrawal of dose drug group, granular cell mitochondrion mild swelling in Figure 56；

7d after the drug withdrawal of dose drug group, pyramidal cell minority mitochondrial swelling, stove cavitation in Figure 57；

1d after the drug withdrawal of dose drug group in Figure 58, glial cell structure is normal, and perivascular space is the most broadening；

1d after the drug withdrawal of dose drug group in Figure 59, synaptic space is the fuzzyyest；

7d after the drug withdrawal of dose drug group in Figure 60, perivascular space is without the most broadening；

1d after Figure 61 high dose medicament group drug withdrawal, the more apparent swelling of granular cell mitochondrion, cavitation；

1d after Figure 62 high dose medicament group drug withdrawal, the more apparent swelling of pyramidal cell mitochondrion, stove cavitation, bite neural thin Born of the same parents' phenomenon；

1d after Figure 63 high dose medicament group drug withdrawal, synaptic space is the fuzzyyest；

1d after Figure 64 high dose medicament group drug withdrawal, perivascular space is the most broadening；

7d after Figure 65 high dose medicament group drug withdrawal, granular cell mitochondrion mild swelling；

7d after Figure 66 high dose medicament group drug withdrawal, pyramidal cell is shown in partial mitochondrial swelling, cavitation；

1d after Figure 67 Normal group drug withdrawal, normal spermatogonium；

7d after Figure 68 Normal group drug withdrawal, normal spermatocyte；

1d after Figure 69 Normal group drug withdrawal, eupyrene sperm；

1d after Figure 70 radiation group drug withdrawal, spermatogonium Chromatin condensation limit is moved；

7d after Figure 71 radiation group drug withdrawal, spermatogonium takes off change；

7d after Figure 72 radiation group drug withdrawal, spermatocyte is downright bad；

1d after Figure 73 radiation group drug withdrawal, spermatocyte mitochondrion obvious tumefaction, cavitation；

1d after Figure 74 radiation group drug withdrawal, spermatid obvious tumefaction, takes off change；

7d after Figure 75 radiation group drug withdrawal, spermatid obvious tumefaction, takes off change；

1d after Figure 76 radiation group drug withdrawal, sperm morphology is normal；

1d after Figure 77 low-dose drugs group drug withdrawal, spermatogonium Chromatin condensation limit is moved；

1d after Figure 78 low-dose drugs group drug withdrawal, spermatogonium Chromatin condensation limit is moved；

1d after Figure 79 low-dose drugs group drug withdrawal, sperm morphology is normal；

1d after Figure 80 low-dose drugs group drug withdrawal, interstitial Ledig cyto-chromatin concentrates limit and moves；

In Figure 81, after the drug withdrawal of dose drug group, 1d, spermatogonium and spermatocyte form are normal, and spermatocyte mitochondrion swells Swollen, cavitation；

In Figure 82, after the drug withdrawal of dose drug group, 7d, spermatogonium and spermatocyte form are normal, and spermatocyte mitochondrion swells Swollen, cavitation；

1d after the drug withdrawal of dose drug group in Figure 83, spermatogonium nuclear chromatin lumps is condensed；

1d after the drug withdrawal of dose drug group in Figure 84, sperm morphology structure is normal；

1d after Figure 85 high dose medicament group drug withdrawal, spermatogonium and spermatocyte form are normal, and spermatocyte mitochondrion swells Swollen, cavitation；

1d after Figure 86 high dose medicament group drug withdrawal, spermatogonium chromatic agglutination limit is moved；

7d after Figure 87 high dose medicament group drug withdrawal, spermatogonium chromatic agglutination lumps coagulation；

7d after Figure 88 high dose medicament group drug withdrawal, spermatocyte mitochondrial swelling, cavitation；

1d after Figure 89 high dose medicament group drug withdrawal, sperm morphology structure is normal；

7d after Figure 90 high dose medicament group drug withdrawal, sperm morphology structure is normal；

The generation in cellular metabolism of Figure 91 active oxygen and eliminating rate.

Detailed description of the invention

Embodiment 1

SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS processing parameter

(1) Thallus Laminariae (Thallus Eckloniae) selects and identifies

The place of production: north of China marine site.

Form: dry bundling.

Identify: identified one by one by SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS quality standard by professional, reject that leaf is yellow, leaf is broken, mouldy etc. no Qualified Thallus Laminariae (Thallus Eckloniae).

(2) operation is soaked

Feed intake: 200 kilograms of Thallus Laminariae (Thallus Eckloniae)s.

Immersion way: in four batches, puts in rustless steel pond for 50 kilograms every batch, is totally immersed into 40 after making Thallus Laminariae (Thallus Eckloniae) unclamp up and down In times amount water.

Soak time: 40 minutes.

(3) clean and shred operation

Cleaning way: rinse Thallus Laminariae (Thallus Eckloniae) to totally with 40 times amount water.

Scavenging period: every batch 10 minutes.

Chopping size: 3 centimetres of big fractionlets of particle diameter.

(4) water extraction process

Take Thallus Laminariae (Thallus Eckloniae) fragment and put into extractor, through successively three steaming and decoctings extraction, add the water of 10 times amount every time, boil for the first time Boiling 2 hours, second and third time respectively boils 1.5 hours.

(5) one is concentrated

Three extracts concentrate on concentration tank to carry out concentrating for the first time.

Mode: concentrating under reduced pressure.

Final concentration: relative density 1.10 (50 DEG C of heat are surveyed).

(6) precipitate with ethanol one

Concentrated solution addition ethanol, to volumetric concentration 30%, carries out precipitating for the first time, stands overnight.

(7) centrifugal one

Collected after centrifugation supernatant, and take sediment sample, carry out assay.[sediment detection] presses SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS matter Amount standard is carried out.

(8) sediment weight is molten

If sediment SO₄ ^2-> 5% time, with the 30% the most molten sediment of ethanol, stand overnight.

(9) it is centrifuged again

SO to sediment₄ ^2-< when 5%, the centrifugal sediment that goes, collection supernatant.

(10) two are concentrated

Centrifuged supernatant concentrates on concentration tank.

Mode: concentrating under reduced pressure.

Final concentration: relative density 1.17 (50 DEG C of heat are surveyed).

(11) precipitate with ethanol two

Concentrated solution addition ethanol, to final concentration 85%, stands overnight.

(12) precipitation is washed

Incline supernatant, adds ethanol to volumetric concentration 75%, washes precipitation, stand overnight.

(13) centrifugal two

Take deposit sample after Li Xin and carry out assay.

(14) it is spray-dried

After Li Xin, add 8 times amount water dissolutioies, be spray-dried, obtain SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS.

Make total amount: 200 kilograms of Thallus Laminariae (Thallus Eckloniae)s obtain 5～6 kilograms of extracts.Obtain 3～5 kilograms after spray drying process to do Powder.Yield 1.5～2.5%.

Confirm that SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS has preventive and therapeutic effect to microwave radiation by following series of experiments.

One, laboratory animal and packet

Two grades of healthy male Wistar rats 210, body weight 200 ± 20g, divides 7 groups at random: Normal group according to body weight (NC), HPM radiation group (HR), low-dose drugs prevents and treats group (LDP), middle dose drug prevents and treats group (MDP), high dose medicament preventing and treating Group (HDP), positive drug group (PD) and middle dose drug matched group (MDC).Laboratory animal is grouped situation in detail and is shown in Table 1.

Table 1 SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS causes Oxidative Injury preventive and therapeutic effect research animal packet situation to HPM

Two, medication

Each group rat 7d 8:00 every day～10:00 gastric infusion before radiation, continue 7d to radiation, be administered 14d altogether.Medicine Thing group drug level is respectively LDP group 2.5mg/ml SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS；MDP group 5mg/ml SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS；HDP group 10mg/ml SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS；PD group 9mg/ml probucol, purchased from Qilu Pharmaceutical Co., Ltd., batch number 4110511KL.The normal saline of the specific volumes such as NC group, HR group all give.MDC group 5mg/ml SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS.It is given daily Before weigh, by the dosage determining every rat when daily weight.

Three, S-band HPM radiation appliance and method of radiating

Use the S-band HPM source that Military Medical Science Institute is self-built, HR group, LDP group, MDP group, HDP group, the radiation of PD group Average power density is 30mW/cm²(frequency: 2.856GHz, peak power density: 200mW/cm², pulse frequency: 300pps, arteries and veins Wide: 500ns), radiated time 60min (radiation 15min, intermittently 15min).Rat is placed in porose lucite box, irradiates Platform the most slowly rotates to ensure that rat body irradiates uniformly.NC group and MDC group carry out false radiation, and other conditions are identical.

Four, rat body weight is measured

Before killing when drawing materials with work before gastric infusion, rat body weight respectively organized by all surveying records every time.

Five, rat immunity system injury index of correlation detection

(1) rat serum routine blood test detection

1. key instrument equipment

Automatic Blood Cell Analyzer, Sysmex 2100, Co., Ltd., Japan.

2. experimental procedure

(1) after drug withdrawal, 1d, 3d, 7d select 10 at random from each group of rat, note through 1% pentobarbital sodium abdominal cavity After penetrating anesthesia, abdomen cardinal vein takes blood 1ml and mixes to anticoagulant tube.

(2) Sysmex 2100 Automatic Blood Cell Analyzer detection rat serum leukocyte count, RBC number, lymph are used Cell number and content of hemoglobin.

(2) rat serum CD3+, CD4+, CD8+ Lymphocyte subtypes test

1. key instrument equipment and reagent

(1) flow cytometer, FACscalibur, BD company, the U.S.

(2) low-temperature and high-speed centrifuge, CR-22E, HITACHI company, Japan

(3) CD3+, CD4+, CD8+ detection kit, BioLegend company, the U.S.

(4) erythrocyte cracked liquid, Hao ocean, Tianjin biological product science and technology limited Company, China

2. experimental procedure

(1) each group rat 1d, 3d, 7d after drug withdrawal, random choose 10, through 1% pentobarbital sodium lumbar injection Anesthesia, takes blood 100 μ l from abdomen cardinal vein, adds FITC anti-rat CD3 antibody 0.5ul, PE after adding heparin sodium anticoagulant Anti-rat CD4 antibody 1.25ul, PerCP anti-rat CD8a antibody 5ul, room temperature places 30min.

(2) often pipe adds erythrocyte cracked liquid 1ml, and room temperature places 5min.

(3) being placed in the refrigerated centrifuge of 4 DEG C by each pipe, 800rpm is centrifuged 5min, abandons supernatant.

(4) add after 500 μ l PBS wash twice, add PBS resuspended.

(5) flow cytomery is used in 1h.

Six, rat brain and the detection of the related oxidized index of testis tissue

1. key instrument equipment and reagent

(1) multi-functional microplate reader, Victor-X3, PerkinElmer company, the U.S.

(2) swirl mixing device, GF-1, Jiangsu its woods medical apparatus factory, China

(3) 96 hole microwell plates, Corning company, China

(4) low-temperature and high-speed centrifuge, CR-22E, HITACHI company, Japan

(5) multi-functional microplate reader, Victor-X3, PerkinElmer company, the U.S.

(6) liquid-transfering gun, Gilson company, France

(7) constant water bath box, Beijing long bearing instruments and meters company, China

(8) XOD, MPO, SOD, GSH-PX, CAT, MDA and PCO detection kit, Nanjing builds up Bioengineering Research Institute, China

2. key step

(1) XOD assay

1. sample pre-treatments: accurately weigh brain and testis tissue weight, by weight volume ratio and add the physiology salt of 9 times of volumes Water makes tissue homogenate, takes appropriate supernatant and measure after being centrifuged.

2. pipe is measured: use Nanjing to build up the XOD testing cassete that article No. is A002 of Bioengineering Research Institute, respectively from each group Sample supernatant samples 10ul add in test tube, sequentially add 1ml reagent one, 0.05ml reagent two, 0.2ml reagent three, 0.02ml reagent four.

3. blank tube: add 10ul distilled water in test tube, sequentially add 1ml reagent one, 0.05ml reagent two, 0.2ml reagent three, 0.02ml reagent four.

4. mix the reagent in each pipe, be placed in water-bath 20min in 37 DEG C of water-baths.

5. taking out each test tube, often pipe is separately added into 1ml reagent five, mixing.

6. taking supernatant, after distilled water zeroing, 530nm, 1cm optical path surveys each pipe absorbance.

(2) MPO assay

1. Sample pretreatment: use Nanjing to build up the MPO testing cassete that article No. is A044 of Bioengineering Research Institute, sampling with 1:1 dilution proportion pressed by reagent two, fully after mixing, accurately takes 0.9ml mixed liquor with pipettor, adds 0.1ml reagent three, mixing Uniformly it is placed on water-bath 15 minutes in 37 DEG C of water-baths.

2. control tube: take 3ml distilled water in test tube, then be separately added into 0.2ml sample, 0.2ml reagent four.

3. pipe is measured: take 0.2ml sample solution in test tube, then be separately added into 0.2ml reagent four, 3ml developer.

4., after mixing the reagent in each pipe, it is placed in water-bath 30min in 37 DEG C of water-baths.

5. taking out each test tube, often pipe is separately added into 0.05ml reagent seven.

6., after mixing the reagent in each pipe, it is placed in water-bath 10min in 60 DEG C of water-baths.

7. take supernatant, after distilled water zeroing, survey each pipe absorbance at 460nm, 1cm optical path immediately.

(3) SOD determination of activity

1. Sample pretreatment: accurately weigh brain and testis tissue weight, by weight volume ratio and add 9 times of normal saline, ice Tissue homogenate is made under water bath condition, 2500 revs/min, after centrifugal 10 minutes, take after taking supernatant normal saline dilution 15 times 50ul measures.

2. control tube: use Nanjing to build up the SOD that article No. is A001-3 (WST-1 method) test of Bioengineering Research Institute Box, addition 0.05ml distilled water is in test tube, then is separately added into 1ml reagent one, 0.1ml reagent two, 0.1ml reagent three and 0.1ml reagent four.

3. measure pipe: add 0.05ml sample solution in test tube, then be separately added into 1ml reagent one, 0.1ml reagent two, 0.1ml reagent three and 0.1ml reagent four.

4., after fully mixing, it is placed in water-bath 40min in 37 DEG C of water-baths.

The most often pipe adds 2ml developer, fully left at room temperature 10min after mixing.

6. take supernatant, after distilled water zeroing, survey each pipe absorbance at 550nm, 1cm optical path immediately.

(4) CAT determination of activity

1. Sample pretreatment: accurately weigh brain and testis tissue weight, adds the physiology of 9 times of volumes according to w/v Saline, makes tissue homogenate under the conditions of ice-water bath, 2500 revs/min, after centrifugal 10 minutes, take supernatant 50ul and measure.

2. control tube: use Nanjing to build up the CAT that article No. is A007-1 (visible ray method) test of Bioengineering Research Institute Box, causes 37 DEG C by reagent one and the pre-temperature of reagent two, is added thereto to 1ml reagent one, 0.1ml reagent two respectively.

3. pipe is measured: addition 0.05ml sample is in test tube, then adds 1ml reagent one, 0.1ml examination to each mensuration pipe respectively Agent two.

4. fully after mixing, 37 DEG C of accurate responses 1 minute.

5. control tube is separately added into 1ml reagent three, 0.1ml reagent four, adds 0.05ml sample solution；Measure in pipe It is separately added into 1ml reagent three, 0.1ml reagent four.

The most fully mix, after distilled water zeroing, survey each pipe absorbance at 405nm, 0.5cm optical path immediately.

(5) GSH-px determination of activity

1. Sample pretreatment: accurately weigh brain and testis tissue weight, by weight volume ratio and add the physiology salt of 9 times of volumes Water makes tissue homogenate, 2500 revs/min, centrifugal 10 minutes, takes supernatant 200ul and measures.

2. control tube adds 1mmol/L GSH 0.2ml, and testing tube adds 1mmol/L GSH and sample to be tested solution is each 0.2ml, 37 DEG C of pre-temperature of water-bath 5 minutes.

3. Nanjing is used to build up the GSH-PX testing cassete that article No. is A005 of Bioengineering Research Institute, by pre-for reagent one temperature extremely 37 DEG C, respectively add 0.1ml reagent one to control tube and testing tube respectively, 37 DEG C of accurate responses 5 minutes.

4. adding 2ml reagent two and 0.2ml sample to be tested solution to control tube, testing tube only adds 2ml reagent two, fully After mixing, 3500 revs/min, centrifugal 10 minutes, take supernatant and carry out subsequent experimental.

1ml GSH standard solvent application liquid, 1ml reagent three, 0.25ml reagent four, 0.25ml is added the most respectively to blank tube Reagent five, standard pipe adds 1ml 20 μm ol/L GSH, 1ml reagent three, 0.25ml reagent four, 0.25ml reagent five, control tube Add 1ml control tube supernatant, 1ml reagent three, 0.25ml reagent four, 0.25ml reagent five, measure pipe and add on 1ml mensuration pipe Clear liquid, 1ml reagent three, 0.25ml reagent four, 0.25ml reagent five.

6. fully after mixing, room temperature stands 15 minutes, after distilled water zeroing, 412nm at, the 1cm optical path each pipe extinction of mensuration Angle value.

(6) MDA assay

1. Sample pretreatment: accurately weigh brain and testis tissue weight, by weight volume ratio and add 9 times of mL normal saline Make tissue homogenate, 2500 revs/min, centrifugal 10 minutes, take appropriate supernatant and measure.

2. using Nanjing to build up the MDA testing cassete that article No. is A003-1 of Bioengineering Research Institute, standard pipe adds 10nmol/ml standard substance 0.2ml and reagent one 0.2ml, adds 3ml reagent two and 1ml reagent three after slight mixing；Blank tube adds Enter 0.2ml dehydrated alcohol and 0.2ml reagent one, after slight mixing, add 3ml reagent two and 1ml reagent three；Mensuration pipe adds 0.2ml sample to be tested and 0.2ml reagent one, add 3ml reagent two and 1ml reagent three after slight mixing.

3., after fully mixing, an aperture, boiling water bath 40 minutes are tightened and pricked on film to test tube mouth preservative film.

4. after flowing water cooling, 3500 revs/min, centrifugal 10 minutes.

5. take supernatant, after distilled water zeroing, survey each pipe absorbance at 532nm, 1cm optical path immediately.

(7) PCO assay

1. the preparation of tissue homogenate: accurately weigh brain and testis tissue weight, by weight volume ratio and add the life of 9 times of volumes Reason saline, makes tissue homogenate, 3000 revs/min, centrifugal 10 minutes, takes supernatant standby under the conditions of ice-water bath.

2. use Nanjing to build up the protein carboxyl-content testing cassete that article No. is A087 of Bioengineering Research Institute, take supernatant 0.225ml, adds 0.025ml protein carbonyl group reagent two, fully mixing room temperature and stands 10min, 11000 revs/min, centrifugal 10min, stays supernatant to be measured.

3. measuring pipe and add 0.1ml sample to be tested and 0.4ml reagent three, control tube adds 0.1ml sample to be tested and reagent Four, it is sufficiently mixed 1 minute, 37 DEG C of accurate lucifuges are reacted 30 minutes.

The most each pipe is separately added into 0.5ml reagent five, is sufficiently mixed 1 minute, is placed in 4 DEG C of refrigerated centrifuges 12000 Rev/min, centrifugal 10min, abandon supernatant, stay precipitation.

The most each pipe is separately added into 1ml dehydrated alcohol ethyl acetate mixing application liquid, is sufficiently mixed 1 minute, is placed on 4 DEG C In refrigerated centrifuge, 12000 revs/min, centrifugal 10min, abandon supernatant, stay precipitation.

6. step 5. three times are repeated.

The most each pipe is separately added into 1.25ml reagent six, after mixing, and 37 DEG C of accurate water-baths 15 minutes.

8. whirlpool mixing, after whole resolution of precipitates, 12000 revs/min, centrifugal 15min.

9., after reagent six zeroing, take supernatant and measure each pipe absorbance at 370nm, 1cm optical path.

Seven, rat hippocampus and testis tissue morphological observation

(1) key instrument equipment

(1) fully-automatic dewatering machine, ASP200, Leica company, Germany

(2) paraffin wax embedding, EG1150, Leica company, Germany

(3) Full-automatic paraffin slice machine, RM2255, Leica company, Germany

(4) cold bench, EG1130, Leica company, Germany

(5) stand sheet machine, HI1220, Leica company, Germany

(6) roasting sheet machine, HI1220, Leica company, Germany

(7) electric drying oven with forced convection, DHG-9145A, Shanghai Yiheng Scientific Instruments Co., Ltd, China

(8) mounting machine, CV5030, Leica company, Germany

(9) optical microscope, DM6000, Leica company, Germany

(10) transmission electron microscope, H7650, HITACHI company, Japan

(2) experiment reagent

(1) haematoxylin dye liquor, Beijing Bioisystech Co., Ltd of Zhong Shan Golden Bridge, China

(2) eosin stain, Beijing Bioisystech Co., Ltd of Zhong Shan Golden Bridge, China

(3) neutral gum, Beijing traditional Chinese medicines group chemical reagent company limited, China

(4) other reagent is domestic analytical pure

(3) experimental procedure

1. light microscopy specimen

(1) after drug withdrawal after 1d, drug withdrawal after 3d, drug withdrawal 7d from each group of rat, select 10 at random, through 1% penta bars After appropriate sodium intraperitoneal injection of anesthesia, take brain and testis tissue, portion of tissue is put in 10% formalin solution fixing.

(2) use graded ethanol as dehydrant, slough the moisture in piece of tissue, then put it in dimethylbenzene transparent.

(3) by tissue input liquid paraffin removes unnecessary dimethylbenzene, piece of tissue is placed in imbedded mold embedding, note Meaning tangent plane direction, adds paraffin and makes it solidify.

(4) wax stone is taken out, be made into thickness and be about the section of 3 μm.Section is put into and drags for sheet machine, after it is smooth, Slice sticker is invested on microscope slide, be placed on roasting sheet machine and dry, make paraffin section.

(5) using dimethylbenzene and graded ethanol to dewax, after haematoxylin dyeing 5-10min, flowing water rinses 3min × 3 Secondary.

(6) acidic alcohol differentiation a moment, flowing water rinses 3min × 3 time.

(7) ethanol Yihong solution is redyed, ethanol dehydration, and dimethylbenzene is transparent, finally uses neutral gum mounting.

(8) section is placed at shady and cool ventilation dries, use observation by light microscope also to take pictures.

2. electron microscope specimen

After drug withdrawal, after 1d, drug withdrawal, after 3d, drug withdrawal, 7d selects 3 at random from each group of rat, through 1% pentobarbital After sodium intraperitoneal injection of anesthesia, take brain and the testis tissue of about 1mm3,2.5% glutaraldehyde and 1% osmic acid and fix, ethanol, acetone Dehydration, epoxy resin embedding, make semithin section location.Then ultrathin section is made, dual through 3% acetic acid uranium and lead citrate After dyeing, observe under transmission electron microscope.

Eight, statistical analysis

Data analysis uses IBM SPSS19.0 software processes, and in literary composition, data are with mean ± standard deviation (mean ± sd) table Show.Each group is compared the variance analysis using repeated measure, and between group, multiple comparisons uses Tukey B method；Remainder data uses Dan Yin Element variance test, multiple comparisons uses LSD method or Dunnett method.Compared with Normal group, * p ＜ 0.05, * * p ＜ 0.01； Compare with HPM radiation group,^ΔP 0.05,^ΔΔP 0.01.Compared with positive drug, * p ＜ 0.05, * * p ＜ 0.01.

Experimental result

One, rat body weight change

Each group rat body weight the most all presents growth trend during gavage, is administered or radiates the growth to rat body weight Have not significant impact.Within 8th day, respectively organizing the more previous celestial body of rat body weight to be heavily basically unchanged, may carry out microwave radiation with the 7th day has Close, see Fig. 1.Prompting 30mW/cm²HPM radiation and gavage give SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS and all have not significant impact rat body weight.

Two, the change of rat immunity index

1. rat leukocyte and lymphocyte number change

The results are shown in Table 1.

(1) 1d after drug withdrawal, radiation group leukocyte count and lymphocyte number compared with normal matched group significantly reduce (P 0.05 or P 0.01).Compared with radiation group, middle dose drug group, positive drug group and middle dose drug matched group leukocyte count are significantly raised (P 0.05), middle dose drug group lymphocyte number significantly raised (P 0.05).Low dosage, high dose medicament group and positive drug Group leukocyte count is significantly lower than Normal group (P significantly lower than Normal group (P 0.05), positive drug group lymphocyte number 0.05).

(2) 3d after drug withdrawal, radiation group leukocyte count and lymphocyte number compared with normal matched group still reduce, and wherein radiation group is white Cell number significantly reduces (P 0.05) compared with matched group.Middle dose drug group and middle dose drug matched group leukocyte count and spoke Penetrating group to compare the most significantly raised (P 0.05), low dosage, high dose medicament group and positive drug group leukocyte count are significantly lower than just Often matched group (P 0.05), each medicine group lymphocyte number no significant difference compared with Normal group.(3) 7d after drug withdrawal, respectively Radiation group, compare between medicine group and Normal group, leukocyte count and the equal no significant difference of lymphocyte number.Prompting, 30mW/ cm²HPM intermittent irradiation 60min can cause rat leukocyte and lymphocyte number is decreased obviously, and gives SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS to HPM Radiogenic rat serum leukocyte and lymphocyte number decline preventive and therapeutic effect.

Table 1 SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS is on rat serum leukocyte and the impact of lymphocyte number after HPM radiation

2. Rat Erythrocytes number and hemoglobin concentration change

The results are shown in Table 2.1d, 3d and 7d after drug withdrawal, each radiation group, compare between medicine group and Normal group, erythrocyte Number and the equal no significant difference of hemoglobin concentration.Prompting, 30mW/cm²HPM intermittent irradiation 60min and gavage give Sargassum sulphuric acid Rat serum RBC number and content of hemoglobin are had no significant effect by polysaccharide or probucol.

Table 2 SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS is on Rat Erythrocytes number and the impact of hemoglobin concentration after HPM radiation

3. rat serum CD3+, the change of CD4+, CD8+ lymphocyte subgroup

Experimental result is shown in Table 3, table 4.Compared with matched group, each radiation group CD3+ and CD4+ lymphocyte 1d after drug withdrawal , though having raised during 7d, all there is not significant change in significantly raised with during 3d (P 0.05).Each radiation group CD8+ lymphocyte With CD4+/CD8+ lymphocyte ratio 1d, 3d and 7d no significant difference after drug withdrawal.Compared with radiation group, low dosage, middle dosage Only 7d significantly raised (P 0.05 or P 0.01) after drug withdrawal with CD3+, CD4+ and CD8+ lymphocyte of high dose medicament group. Compared with Normal group, each medicine group CD3+ and CD4+ lymphocyte significantly raised (P when 1d, 3d and 7d after drug withdrawal 0.05)；Positive drug group CD8+ lymphocyte significantly raised during 1d after drug withdrawal (P 0.05), low dosage, middle dose drug group CD8+ lymphocyte significantly raised during 7d after drug withdrawal (P 0.05)；Each medicine group CD4+/CD8+ lymphocyte ratio is in drug withdrawal During rear 3d significantly raised (P 0.05).This experiment experimental result different from preliminary result, find out from data, CD3+ with The radiation group of CD4+ lymphocyte all occurs in that rising in various degree with medicine group compared with matched group, it may be possible to due to microwave The irritability reaction that radiation causes is caused.

Table 3 SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS is to rat serum lymphocyte CD 3 after HPM radiation⁺、CD4⁺Impact

Table 4 SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS is to rat serum lymphocyte CD 8 after HPM radiation⁺And CD4⁺/CD8⁺The impact of ratio

Three, the change of rat different organs oxidative stress index

1.XOD activity

Experimental result is shown in Table 5.(1) 1d after drug withdrawal, radiation group cerebral tissue XOD activity with matched group than more significant rising (P ＜ 0.01), other medicines group significantly raised compared with radiation group (P ＜ 0.05), low-dose drugs group in addition to low-dose drugs group XOD activity with the obvious rising of matched group (P ＜ 0.05).The XOD activity of radiation group testis tissue compares bright with matched group Aobvious rising (P ＜ 0.05), XOD activity and the obvious rising of matched group (P ＜ 0.05), the middle dose drug group of positive drug group Substantially (P ＜ 0.05) is lowered compared with radiation group, middle dose drug group and the comparison of middle dose drug with middle dose drug matched group Group the most substantially reduces (P ＜ 0.05) compared with positive drug group, shows that it reduces the XOD activity elevating effect ratio sun that HPM causes Property medicine is more preferable.(2) 3d after drug withdrawal, radiation group, compares between medicine group and Normal group, and brain, liver, spleen, the XOD of testis live The equal no significant difference of property.(3) 7d after drug withdrawal, radiation group compares with Normal group, and the XOD activity of brain and testis is without the poorest Different；The XOD activity of middle dose drug group, high dose medicament group, positive drug group and middle dose drug matched group cerebral tissue and spoke Penetrate group and Normal group compares the most substantially reduction (P ＜ 0.05), middle dosage, the XOD activity of high dose medicament group testis tissue Relatively radiation group substantially reduces (P ＜ 0.05).

2.MPO activity

Experimental result is shown in Table 5.(1) 1d after drug withdrawal, the MPO activity of radiation group brain and testis tissue compares with Normal group The most significantly raised (P ＜ 0.05)；The MPO of middle and high dose drug group, positive drug group and middle dose drug matched group cerebral tissue lives Property substantially reduce (P ＜ 0.05) compared with radiation group, the MPO activity of dose drug group testis tissue low, middle is compared with matched group Significantly reducing (P ＜ 0.05), the MPO of each medicine group brain and testis tissue active comparison with Normal group has no significant change. (2) 3d after drug withdrawal, MPO activity significantly raised compared with Normal group (P ＜ 0.05) of radiation group brain, the MPO of each medicine group Activity the most substantially reduces (P ＜ 0.05 or P ＜ 0.01) compared with radiation group, compares without significant change with Normal group；Radiation The MPO activity of group testis tissue compares with Normal group does not has significant change, high dose medicament group and positive drug group and spoke Penetrate group and compare substantially reduction (P 0.05), without significant change compared with Normal group.(3) 7d after drug withdrawal, radiation group, medicine group And compare between Normal group, MPO activity all no significant differences of brain and testis.

Table 5 SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS is on rat different organs XOD, the impact of MPO activity after HPM radiation

3.SOD activity

Experimental result is shown in Table 6.(1) the SOD activity of 1d after drug withdrawal, radiation group brain and testis tissue is obvious compared with matched group Reduce (P ＜ 0.05).SOD activity (P the ＜ 0.05 or P ＜ the most significantly raised compared with radiation group of each medicine group cerebral tissue 0.01), in addition to low-dose drugs group, the SOD activity of each medicine group testis tissue significantly raises (P ＜ compared with radiation group 0.01).Each medicine group cerebral tissue SOD activity compared with Normal group without significant change, but the SOD of testis tissue activity and Normal group compares the most significantly raised (P ＜ 0.05 or P ＜ 0.01).(2) 3d after drug withdrawal, radiation group brain and testis tissue SOD activity the most substantially reduces (P ＜ 0.05) compared with Normal group, the SOD activity of middle dose drug group brain and testis tissue Compared with radiation group significantly raised (P ＜ 0.05), the SOD activity compared with normal matched group of each medicine group brain and testis is without the poorest Different.(3) 7d after drug withdrawal, the SOD activity of radiation group cerebral tissue compares the most substantially reduction (P ＜ 0.05), middle dosage medicine with matched group Thing group significantly raised compared with radiation group (P 0.05)；The SOD activity of testis tissue after irradiation compared with Normal group not There is significant change, the SOD activity equal no significant difference of compared with normal matched group of each medicine group brain and testis.

4.GSH-px activity

Experimental result is shown in Table 6.(1) 1d after drug withdrawal, the GSH-px activity compared with normal matched group of radiation group cerebral tissue does not finds Significant change, low, high dose medicament group the most significantly raised (P ＜ compared with Normal group, radiation group, positive drug group 0.05).Each radiation group, compare between medicine group and Normal group, testis tissue GSH-px activity all no significant differences.(2) 3d after drug withdrawal, the GSH-px activity compared with normal matched group of radiation group cerebral tissue does not finds significant change yet, middle dose drug group and Middle dose drug matched group the most significantly raised compared with Normal group, radiation group, positive drug group (P ＜ 0.05).Each radiation Group, compare between medicine group and Normal group, testis tissue GSH-px activity all no significant differences.(3) 7d after drug withdrawal, radiation The GSH-px activity compared with normal matched group of group cerebral tissue does not finds significant change, high dose medicament group and radiation group and positive drug yet Thing group compares the most significantly raised (P ＜ 0.05).Each radiation group, compare between medicine group and Normal group, testis tissue GSH- Px activity all no significant differences.

Table 6 SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS is on rat different organs SOD, the impact of GSH-px activity after HPM radiation

5.CAT activity

Experimental result is shown in Table 7.(1) 1d after drug withdrawal, radiation group testis tissue CAT activity substantially drops compared with Normal group Low (P ＜ 0.05), the CAT activity relatively radiation group of middle and high dose drug group and positive drug group is the most significantly raised (P ＜ 0.05), But all there is not significant change compared with Normal group and positive drug group in each medicine group.Each radiation group, medicine group and normal Compare between matched group, cerebral tissue CAT activity all no significant differences.(2) 3d after drug withdrawal, radiation group cerebral tissue CAT activity with Normal group compares does not has significant change, the CAT activity compared with normal matched group of middle and high dose drug group and positive drug group, Radiation group the most significantly raised (P ＜ 0.05).The CAT activity of radiation group testis tissue the most substantially reduces (P compared with Normal group ＜ 0.05), high dose medicament group, positive drug group the most substantially rise compared with radiation group with middle dose drug matched group CAT activity High (P ＜ 0.05 or P 0.01), middle dose drug matched group CAT activity is the most obvious compared with Normal group, positive drug group Raise (P ＜ 0.05).(3) 7d after drug withdrawal, each radiation group, compares between medicine group and Normal group, brain and testis tissue CAT Activity all no significant differences.

Table 7 SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS is on the impact (unit: U/ml) of rat different organs CAT activity after HPM radiation

6.MDA content

Experimental result is shown in Table 8.(1) 1d after drug withdrawal, compares with Normal group, and the MDA content of radiation group cerebral tissue is obvious Increasing (P ＜ 0.05), the MDA content of dose drug group cerebral tissue low, middle the most substantially reduces (P ＜ 0.05 or P compared with radiation group ＜ 0.01), the MDA content compared with normal matched group of middle dose drug group cerebral tissue, positive drug group the most substantially reduce (P ＜ 0.05).The MDA content of radiation group testis tissue substantially increases (P ＜ 0.05) compared with Normal group, except low-dose drugs group The MDA content of outer each medicine group testis compares the most substantially reduction (P ＜ 0.05) with radiation group, and with Normal group without the poorest Different.(2) 3d after drug withdrawal, the MDA content of radiation group cerebral tissue dramatically increases (P ＜ 0.01) compared with Normal group, except low dose The MDA content of the outer each medicine group brain of amount medicine group the most substantially reduces (P ＜ 0.05) compared with radiation group.Radiation group testis tissue MDA content substantially increases (P ＜ 0.05) compared with Normal group, and the MDA content of each medicine group testis is equal compared with radiation group Substantially reduce (P ＜ 0.05), and with Normal group no significant difference.(3) 7d after drug withdrawal, the MDA content of radiation group cerebral tissue Compare with matched group the most significantly raised (P ＜ 0.05), middle dose drug group, positive drug group and middle dose drug matched group and spoke Penetrate group to compare and substantially reduce (P ＜ 0.05 or P ＜ 0.01), and with Normal group no significant difference.Each radiation group, medicine group and Compare between Normal group, the equal no significant difference of testis tissue MDA content.

7.PCO content

Experimental result is shown in Table 8.(1) 1d after drug withdrawal, the PCO content of radiation group cerebral tissue and the obvious increasing of Normal group Adding (P ＜ 0.05), in addition to low-dose drugs group, the PCO content of each medicine group brain the most substantially reduces (P compared with radiation group 0.05), and with Normal group no significant difference.The PCO content of radiation group testis tissue with Normal group than more significant increasing Adding (P ＜ 0.01), in addition to high dose medicament group, the PCO content of each medicine group testis the most substantially reduces (P compared with radiation group 0.05 or P 0.01), and with Normal group no significant difference.(2) 3d and 7d after drug withdrawal, each radiation group, medicine group and normal Compare between matched group, brain and the equal no significant difference of testis tissue PCO content.

Table 8 SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS is on rat different organs MDA, the impact of PCO content after HPM radiation

Four, rat hippocampus and Testis Morphology change

1. histological change

(1) hippocampal tissue change

1d-7d after Normal group drug withdrawal, rat brain hippocampal tissue overwhelming majority neuron morphology structure is normal, accidental Single neuron pyknosis, deeply contaminate, perivascular space the most broadening (Fig. 2-3).30mW/cm²7d-14d after HPM intermittent irradiation 1h, Hippocampal tissue change pathological changes basic simlarity, 7d (1d after drug withdrawal) after radiation, pyknosis, the neuron compared with normal matched group of deep dye increase Many, cell and perivascular space are also broadening compared with matched group, cellular edema (Fig. 4)；10d (3d after drug withdrawal), above-mentioned damage after radiation Hindering degree to increase the weight of further, the neuronal quantity that karyopycnosis contaminates deeply substantially increases, and cellular edema increases the weight of, cell and perivascular space The most broadening (Fig. 5)；14d (7d after drug withdrawal) after radiation, neuron pyknosis is deeply infected with and is recovered, but still sees that partial nerve unit core is solid The deep dye of contracting, and cell and broadening the having no of perivascular space substantially alleviate (Fig. 6).

After low-dose drugs group drug withdrawal, 1d hippocampal tissue pathological changes is similar to radiation group, but 3d-7d has alleviated (Fig. 7-9)； Middle dose drug group pathological changes relatively radiation group substantially alleviates, 1d and 3d after drug withdrawal, sees being dispersed in single neuron pyknosis, deeply contaminate, cell The most relatively radiation group broadening with perivascular space alleviates, cellular edema (Figure 10-11), 7d after drug withdrawal, and basic recovery normally (is schemed 12).High dose medicament group pathological changes is similar to middle dose drug group, but relatively middle dose drug group is slightly heavy (Figure 13-14), 7d after drug withdrawal Also basic recovery is normal (Figure 15).

Positive drug group hippocampal tissue pathological changes similar to high dose medicament group (Figure 16-17), but after drug withdrawal, 7d is the most complete Recover normal (Figure 18).Middle dose drug matched group, 1-7d after drug withdrawal, hippocampal tissue has no obvious pathological changes (Figure 19-20).

(2) testicular histology's change

Rats in normal control group testis spermatogenic cells at different stages has levels arrangement, and seminiferous tubule limitans is complete, intracavity sperm count Amount is abundant (Figure 21).30mW/cm²After HPM intermittent irradiation 1h, 7d (1d after drug withdrawal), 10d (3d after drug withdrawal) and 14d are (after drug withdrawal 7d), testis tissue change basic simlarity, but gradually alleviate, in showing as testis periphery seminiferous tubule, spermatogenic epithelium layer is obvious Edema, spermatogenic cell not close-packed arrays, spermatid and sperm quantity reduce (Figure 22) in various degree, part seminiferous tubule intracavity And albumen edematous fluid accumulation (Figure 23-24) seen from interstitial, spermatogenic cell degeneration, necrosis seen from minority seminiferous tubule and come off.

Low-dose drugs group and radiation group lesion characteristic, rule and degree basic simlarity (Figure 25～27)；Middle dose drug Group pathological changes relatively radiation group substantially alleviates, 1d and 3d after drug withdrawal, it is seen that be dispersed in the seminiferous tubule light intermediate edema of spermatogenic epithelium layer, spermatogenesis Cell arrangement is the most loose, has no obvious spermatid and oligospermia (Figure 28-29), accidental single seminiferous tubule intracavity and focal Property interstitial seen from albumen edematous fluid accumulation, have no spermatogenic cell degeneration, necrosis and come off；7d after drug withdrawal, basic recovery normally (is schemed 30).High dose medicament group pathological changes is similar to middle dose drug group, but relatively middle dose drug group is slightly heavy (Figure 31-32), 7d after drug withdrawal Recover normal (Figure 33) the most completely.

Positive drug group testis tissue pathological changes similar to high dose medicament group (Figure 34-35), but than the heaviest, 7d after drug withdrawal Recover normal (Figure 36) the most completely.Middle dose drug matched group, after drug withdrawal 1～7d, testis tissue has no significant change (figure 37-38)。

3. Change of Ultrastructure

(1) Hippocampal ultrastructure change

Rats in normal control group Hippocampus granular cell and pyramidal cell neuron and glial cell object line plastochondria slightly swell Swollen, endoplasmic reticulum is slightly expanded, and synaptic space is the fuzzyyest, and perivascular space normal or the most broadening (Figure 39-43), except indivedual colloids Cell Mild edema is outer (Figure 44), and remaining shows no obvious abnormalities.

30mW/cm²HPM is interrupted 7d (1d after drug withdrawal) and 14d (7d after drug withdrawal), hippocampal neuron nuclear chromatin after irradiation Loose, mitochondrion obvious tumefaction, cavitation, lysosome increasing number (Figure 45)；Glial cell obvious tumefaction, endochylema organelle is sparse (Figure 46), synaptic space is fuzzy (Figure 47)；Perivascular space the most broadening (Figure 48).

1d after low-dose drugs group drug withdrawal, hippocampal tissue Change of Ultrastructure is similar to radiation group, but than the heaviest.Granule The mitochondrial swelling of cellular neural unit, cavitation, lysosome quantity increases, and endoplasmic reticulum and Golgi body are slightly expanded (Figure 49), or Granulocyte obvious tumefaction, takes off change (Figure 50), or chromatic agglutination limit is moved, in apoptosis (Figure 51)；Pyramidal cell mitochondrion is bright , there is neuronophagia (Figure 52) in aobvious swelling.Synaptic space obscures (Figure 53)；Perivascular space moderate broadening (Figure 54).

1d and 7d after middle dose drug group drug withdrawal, hippocampal tissue Change of Ultrastructure relatively radiation group substantially alleviates, and normally Matched group no significant difference.Granular cell neuron minority mitochondrion mild swelling (Figure 55-56), pyramidal cell minority mitochondrion More apparent swelling, stove cavitation (Figure 57), glial cell morphosis is normal (Figure 58), and synaptic space is slightly fuzzy (Figure 59), blood Pipe week gap the most broadening or normal (Figure 58,60).

1d and 7d after high dose medicament group drug withdrawal, similar to middle dose drug group, but the relatively middle dose drug group of 1d is the heaviest, The more apparent swelling of granulocyte neuron linear plastochondria, cavitation (Figure 61)；The more apparent swelling of pyramidal cell mitochondrion, cavitation, endoplasmic reticulum Expand in various degree with Golgi body, neuronophagia (Figure 62) occurs；Synaptic space is slightly fuzzy (Figure 63)；Between blood vessel week Gap the most broadening (Figure 64).After drug withdrawal, the above-mentioned pathological changes of 7d substantially alleviates, and granular cell and pyramidal cell are shown in minority or partial line grain Body swelling, cavitation (Figure 65-66), glial cell and perivascular space recover normal substantially.

(2) testis Change of Ultrastructure

1d and 7d after Normal group drug withdrawal, rat testicle seminiferous tubule structure is normal, and a small amount of spermatogenic cell, Leydig are thin Born of the same parents and the sertoli cell slight swelling of object line plastochondria, endoplasmic reticulum and Golgi body mild dilation (Figure 67-69).

30mW/cm²7d (1d after drug withdrawal) and 14d (7d after drug withdrawal) after HPM interruption irradiation, spermatogonium peripheral edema, carefully Intercellular space is broadening, and (Figure 70) is moved on nuclear chromatin condensation, limit, and nuclear membrane obscures, takes off change (Figure 71), mitochondrion moderate swelling, cavitation；Essence Daughter cell and spermatocyte mitochondrial swelling, cavitation are obvious, reticulum dilatation, it is seen that spermatocyte and spermatid take off change and bad Extremely (Figure 72-75)；Mature sperm head form normal (Figure 76).

1d after low-dose drugs group drug withdrawal, testis tissue Change of Ultrastructure is similar to radiation group, but degree is the lightest.Essence is former (Figure 77) is moved on the cohesion of nuclei dyeing chromaticness, limit, and mitochondrion is mild swelling, stove cavitation；Spermatocyte and spermatid line grain Body obvious tumefaction, cavitation (Figure 78), have no that spermatocyte and spermatid take off change and necrosis；Mature sperm head form is the most normal (Figure 79)；But (Figure 80) is moved on visible interstitial Leydig cyto-chromatin coagulation limit.

1d and 7d after middle dose drug group drug withdrawal, most of spermatogonium structures recover normal substantially, and only a small amount of cell can See the slight swelling of mitochondrion, spermatocyte and the slight swelling of spermatid mitochondrion, cavitation, but relatively radiation matched group alleviates (figure 81-82), accidental spermatogonium nuclear chromatin condensation (Figure 83), have no that spermatocyte and spermatid are downright bad；Mature sperm head Form normal (Figure 84).Sertoli cell and interstitial Leydig cellularity are normal, and accidental mitochondrion slightly swells Swollen, endoplasmic reticulum mild dilation.

1d and 7d after high dose medicament group drug withdrawal, similar to middle dose drug group pathological changes, but than the heaviest, most of essences are former Cellularity is normal, it is seen that mitochondrion mild swelling, and endoplasmic reticulum is slightly expanded (Figure 85), minority spermatogonium nuclear chromatin (Figure 86-87) is moved on cohesion, limit, and part spermatocyte and the more apparent swelling of spermatid mitochondrion, cavitation (Figure 85,88) have no Spermatocyte and spermatid are downright bad；Mature sperm head form normal (Figure 89-90).Sertoli cell and interstitial Leydig cellularity is normal, it is seen that the slight swelling of partial mitochondrial, endoplasmic reticulum mild dilation.

By experimental studies have found that, it is the most sensitive that brain and testis cause oxidativestress damage to high power microwave radiation.Brain exists Need more energy during vital movement, energy consuming process can produce a certain amount of ROS, and corresponding antioxidase Content is the most relatively low, so that it is vulnerable to the attack of ROS；Cerebral tissue neuron membrane structure, rich in polyunsaturated fatty acid, is easily subject to Attack to body free radical；Ca in neuron²⁺Circulation is higher, when being easily caused intracellular calcium overload by environmental damage, this A little factors all can cause the central nervous system oxidation stress.And testis blood supply is the poorest, and sexual cell propagation, Differentiation capability is relatively strong, and DNA constantly carries out the process such as uncoiling, duplication, makes energy metabolizing enzyme, apoptosis-related protein, DNA Deng the impact easily receiving various chemical factors.Therefore, testis tissue is easily by microwave radiation damage.

One, SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS causes brain and testicular histology and the preventive and therapeutic effect of ultrastructure damage to microwave radiation

Morphological changes of various tissue components be research aircraft soma, organ by degree of injury after environmental stimuli must obligato part, There is higher accuracy and objectivity.Ma Di etc. use 30mW/cm²Microwave radiation Wistar rat 15min, finds after 14d Major part hippocampal neuron karyopycnosis, deeply contaminate, perivascular space is the most broadening；Find in ultrastructure that neuron linear plastochondria swells Swollen, cavitation, ridge fracture or dissolving, synaptic vesicle accumulation, synaptic space is smudgy.Lv Shijie etc. are respectively adopted 100mW/cm²、 200mW/cm²Microwave radiation Wistar rat 5min, finds 100mW/cm²Organize 6h group neuronal damage after irradiation relatively light, 48h Rear endochylema vacuolation, mitochondrial swelling, cavitation.200mW/cm²6h after radiation, nuclear membrane is coarse, and mitochondrial swelling, cavitation are obvious； After 48h, nuclear membrane ruptures, and mitochondrial crista lacks, chromatolysm.Guo Guozhen etc. use 2450MHz microwave radiation BALB/c mouse testis Ball 15min, finds spermatogenic cell cloudy swelling in various degree, downright bad and come off, spermatogonium edema, spermatocyte degeneration, comes off, Sperm quantity reduces.

Experimental result of the present invention is similar with the above results, 30mW/cm²HPM radiation cause hippocampal neuron pyknosis, deep dye and Edema, perivascular space is the most broadening；The obvious edema of spermatogenic epithelium layer, spermatid and essence in testis periphery seminiferous tubule Son reduces in various degree；Ultrastructure is shown in radiation group hippocampal neuron and glial cell edema, mitochondrion obvious tumefaction, cavitation, Synaptic space obscures, and perivascular space is the most broadening；Testicle spermatogonia nuclear chromatin condensation, limit are moved or take off change, spermatocyte With spermatid mitochondrion obvious tumefaction, cavitation, it is seen that spermatocyte and spermatid take off change and necrosis.After microwave radiation, 7d can Observing above-mentioned change, after radiation, 10d damage increases the weight of, and has alleviated after 14d.

The present invention test result indicate that, microwave radiation is caused each dose drug group Hippocampus and injury of testis all has certain controlling Treatment effect.Wherein, 50mg/kg/d medicine group preventive and therapeutic effect effect is best.

Two, SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS causes brain and the preventive and therapeutic effect of testis oxidativestress damage to microwave radiation

Active oxygen is the intermediate product that organism metabolism is movable, and it can attack lipid, protein and DNA etc., causes the third two Aldehyde, content of protein carbonyl group raise, DNA break, and then cause histiocyte impaired；Can also be with antioxidases such as SOD, CAT Effect, reduces activities of antioxidant enzymes.Therefore, the present invention is by detection brain, testis tissue oxidative stress intermediates content and The activity of a little antioxidases, generates the order of relevant enzyme-antioxidase-Radical Metabolism product according to free radical, systematically from The angle of oxidative stress evaluates microwave radiation to brain, the damage effect of testis tissue.

(1) microwave radiation causes brain and the impact of testis free radical generation enzymatic activity

Xanthine oxidase (XOD) is the metabolic enzyme of body nucleotide, and it can be catalyzed hypoxanthine and generate xanthine, enters One step generates uric acid；Can also directly be catalyzed xanthine generate uric acid, and along with ultra-oxygen anion free radical, hydroperoxyl radical and The generation of hydrogen peroxide.Myeloperoxidase (MPO) (MPO) is the hemoprotein enzyme of prosthetic heme group, be present in myeloid cell addicted to In aniline blue granule.MPO can utilize hydrogen peroxide and chloride ion to produce hypochlorite, and forms the freedom with oxidability Base.As can be seen here, XOD and MPO plays an important role in the generation of free radical.

In the experiment of the present invention, cerebral tissue XOD (1d after radiation), MPO (1d, 3d after radiation) and testis tissue XOD (radiation Rear 1d), MPO (1d after radiation) the most significantly raised, show HPM can cause part free radical generate related enzyme activity raise, enter And cause body free-radical contents to raise.

(2) microwave radiation cause brain becomes the impact of enzymatic activity with testis antibiosis

Superoxide dismutase (SOD) is the potential inducer of body oxidative damage and in multiple physiology and pathological process In play important role, such as apoptosis, cell cycle arrest etc., it can be removed superoxide radical, reduce it The damage that body is caused.Glutathion peroxidase (GSH-PX) acts primarily as detoxification, major function in organism It is that lipid peroxide is reduced to nontoxic hydroxy compounds, the hydrogen peroxide that reduction is free, it is catalyzed glutathion simultaneously, from And protect membrane structure and fully functional.Catalase (CAT) is a kind of enzyme being widely present in different kind organism body, is Antioxidizing agent, its effect is that catalyzing hydrogen peroxide is converted into water and oxygen, prevents it from causing damage body.Active oxygen (ROS) eliminating rate in cellular metabolism is as shown in Figure 91.

In the experiment of the present invention, cerebral tissue SOD (1d, 3d, 7d after radiation) and testis tissue SOD (1d, 3d after radiation), CAT (1d, 3d after radiation) the most substantially reduces, and shows that HPM can reduce Partial Antioxidation enzymatic activity, causes body ROS to remove speed Rate reduces.

(3) microwave radiation is on brain and the impact of testis Radical Metabolism product assay

Malonaldehyde (MDA) is the end-product of body lipid peroxidation, can cause the polymerization such as protein, nucleic acid, Aggravation membrane damage also affects key enzyme activity in mitochondrial respiratory chain and mitochondrion, and the change of MDA content can indirect reaction body The change of free-radical contents.Protein carbonyl group (PCO) is the product of protein oxidative damage, is to evaluate the total oxidation level of protein Common counter.Because body protein is of a great variety, widely distributed, therefore be highly prone to active oxygen and attack, cause protein to be tied Structure and the change of conformation, ultimately result in protein inactivation.

In the experiment of the present invention, cerebral tissue MDA (1d, 3d, 7d after radiation), PCO (1d after radiation) and testis tissue MDA (1d, 3d after radiation), PCO (1d after radiation) are the most significantly raised, show that HPM can increase the content of body free radical product, increase Add the damage to body.

(4) SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS to microwave radiation to brain and the preventive and therapeutic effect of testis oxidativestress damage

The experiment of the present invention is by using three kinds of various dose SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS to test, and result shows middle dosage medicine The preventive and therapeutic effect of thing group (50mg/kg/d) is best, is mainly manifested in cerebral tissue XOD activity (1d, 7d after drug withdrawal), MPO activity and (stops 1d, 3d after medicine), MDA content (1d, 3d, 7d after drug withdrawal) and PCO content (1d after drug withdrawal) relatively radiation group substantially reduce, SOD activity (1d, 3d, 7d after drug withdrawal) is significantly raised.Testis tissue XOD activity (1d after drug withdrawal), MPO activity (1d after drug withdrawal), MDA content (1d, 3d after drug withdrawal) and PCO content (1d after drug withdrawal) relatively radiation group substantially reduce, SOD activity (1d, 3d after drug withdrawal), CAT activity (1d after drug withdrawal) is significantly raised.

By this part experimentation, we specify that effective to HPM radiation damage preventive and therapeutic effect of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS Dosage.Meanwhile, SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS can reduce the free radicals such as XOD, MPO and generate related enzyme activity, increases the antioxidation such as SOD, CAT Enzymatic activity, reduces the Radical Metabolism product assay such as MDA, PCO, one of mechanism of body oxidative damage caused by its preventing and treating HPM.

Experiment conclusion:

One, 30mW/cm²HPM radiation may result in immune functions of rats and reduces, and mainly shows as leukocyte and lymphocyte number Amount declines, and other immune indexes is without significant change.

Two, 30mW/cm²HPM radiation can cause the morphology damage in various degree of rat brain and testis, is mainly manifested in Neuron pyknosis contaminates deeply, spermatogenic epithelium cellular edema.Each medicine group can alleviate above-mentioned damage, and wherein, middle dose drug group is imitated The most best.

Three, 30mW/cm²HPM radiation can cause rat brain and testis oxidativestress damage in various degree, mainly shows Raising in MPO and XOD activity, SOD and CAT activity reduces, MDA and PCO content raises.

Four, the SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS of each dosage all can prevent and treat 30mW/cm to some extent²HPM radiation causes rat and exempts from Epidemic disease function reduces and the oxidative damage of rat different organs.Wherein, middle and high dose drug group preventive and therapeutic effect is close, preventing of the two Control effect more preferable than low-dose drugs group effect, it was demonstrated that middle dosage (50mg/kg/d) medicine group is its optimal administration concentration.

Five, some Testing index of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS middle and high medicine group is better than probucol positive drug group, shows that it has There is good antioxidation.

Claims

1. the application of a SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS, it is characterised in that described SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS is in terms of preventing and treating microwave radiation Application.

The application of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS the most according to claim 1, it is characterised in that described sulphuric acid Sargassum polysaccharides is in preparation Application on the medicine of preventing and treating microwave radiation.

The application of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS the most according to claim 1 and 2, it is characterised in that microwave in described microwave radiation For the S-band microwave that frequency 2-4GHz, wavelength are 150-75.00mm.

The application of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS the most according to claim 3, it is characterised in that described microwave is frequency 2.856GHz Microwave.

The application of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS the most according to claim 4, it is characterised in that described microwave average power density is 30mW/cm², peak power density is 200mW/cm², pulse frequency is 300pps, and pulsewidth is 500ns.

The application of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS the most according to claim 1, it is characterised in that the extracting method of described Sargassum polysaccharides Comprise the following steps:

(1) after Thallus Laminariae (Thallus Eckloniae) being placed in the water of himself quality 38-42 times immersion 35-45 minute, for himself quality The water of 38-42 times amount is rinsed well；

(2) Thallus Laminariae (Thallus Eckloniae) is carried out pretreatment, reject the defective Thallus Laminariae (Thallus Eckloniae) that leaf is yellow, leaf is broken and mouldy, qualified Thallus Laminariae (Thallus Eckloniae) is cut into area 7- The fragment of 10 square centimeters；

(3) Thallus Laminariae (Thallus Eckloniae) is joined and decoct three times in the water of himself quality 9-11 times, respectively 1.9-2.1 hour, 1.4- 1.6 hours, 1.4-1.6 hour；

(5) in concentrated solution I, ethanol is added to ethanol volumetric concentration 29-31%, centrifugal supernatant and heavy after standing overnight Slag；

(6) sediment is detected, if SO in sediment₄ ^2-Weight/mass percentage composition more than 5%, in sediment, add ethanol to second Alcohol volumetric concentration 29-31%, stands to layering and is centrifuged to obtain supernatant；If SO in sediment₄ ^2-Weight/mass percentage composition be less than In 5%, abandon sediment；

(7) supernatant in step S50 and step S60 is merged, be evaporated to relative density 1.1-1.2 and obtain concentrated solution II；

(8) in concentrated solution II, addition ethanol is to ethanol volumetric concentration 83-86%, after standing overnight, abandons supernatant, and precipitation is used It is centrifuged after the washing with alcohol of percentage by volume 74-76%；

The application of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS the most according to claim 6, it is characterised in that

After Thallus Laminariae (Thallus Eckloniae) being placed in the water of himself quality 40 times by step (1) immersion 40 minutes, for himself quality 40 times The water of amount is rinsed well；

In step (3), Thallus Laminariae (Thallus Eckloniae) is joined in the water of himself quality 10 decoct three times, respectively 2 hours, 1.5 hours, 1.5 hour；

In step (5), in concentrated solution I, add ethanol to ethanol volumetric concentration 30%, after standing overnight centrifugal supernatant and Sediment；

In step (6), if SO in sediment₄ ^2-Weight/mass percentage composition more than 5%, in sediment, add ethanol dense to ethanol volume Degree 30%, stands to layering and is centrifuged to obtain supernatant.

In step (7), the supernatant in step S50 and step S60 is merged, be evaporated to relative density 1.17 and obtain concentrated solution II。

In step (8), in concentrated solution II, addition ethanol is to ethanol volumetric concentration 85%, after standing overnight, abandons supernatant, precipitation With centrifugal after the washing with alcohol of percentage by volume 75%.