CN100391472C - Use of testiset and its composition in health-care products for preventing liver from chemical damage - Google Patents
Use of testiset and its composition in health-care products for preventing liver from chemical damage Download PDFInfo
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Abstract
The present invention discloses an application of seal's fat containing effective components of docosahexenoic acid, icosapent and docosa-carbon pentaene acid for preparing health products for controlling chemical liver injury. The present invention discloses the application of a seal's fat composition which is composed of 99.5 wt% of seal's fat and E0.5 wt% of vitamin for preparing health products for controlling chemical liver injury. Experiments prove that the product has auxiliary protective action to chemical liver injury when the product is administrated for a long time.
Description
Technical field
The present invention relates to the application in preparation control chemical liver injury health product of a kind of Adeps Phocae vitulinae and compositions thereof, belong to field of health care products.
Background technology
Fur seal is being commonly called as of Phocidae animal harp sea dog (Harp seal), and perch is in the arctic and Atlantic the north, and it is lived in the environment that does not almost have to pollute, and temperature on average is at subzero 20 ℃～40 ℃.What fur seal was different with Trionyx sinensis Wiegmann is, it is a warm-blooded animal, compare with the mammal sheep, it is because live in the characteristic that has yin nourishing in the water again, so fur seal is the animal that has YANG invigorating animal such as sheep and yin nourishing animal such as Trionyx sinensis Wiegmann characteristic concurrently, therefore it has the function of YANG invigorating and yin nourishing concurrently, fur seal property is flat, and salty in the mouth is gone into the warp of Liver and kidney, having YIN nourishing and moisten the merit of positive kidney-reinforcing Yang-strengthening, is ideal tonic food.
Because the protective measure that Canadian government is taked, the quantity of fur seal increases with annual 7% velocity-stabilization.For keeping the ecological balance between the fur seal and the shoal of fish, through the United Nations and Canadian government approval, can catch and kill fur seal every year slightly.Like this, extract in the fur seal body various nutritional labelings for human service has just had may.
Fatty acid is closed in many carbon of ω-3 type insatiable hunger of being rich in needed by human in the Adeps Phocae vitulinae, and (ω-3PUFA), its nutritive value is better than other edible oils.Outside the eicosapentaenoic acid of in Adeps Phocae vitulinae, finding to contain usually in the fish oil (EPA) and docosahexenoic acid (DHA), also has the considerable clupanodonic acid (DPA) of content.
Present human hepatopath is more and more, and has special positive effect and the medicine of almost being free from side effects is difficult to confirm.Wherein, hepatocyte injury may be the result of multiple factor interaction.The mechanism of hepatic injury thinks relevant with the lipid peroxidation process of intrahepatic fat acid metabolic obstacle and free radical and initiation thereof at present more due to the chemical substance (as carbon tetrachloride, ethanol etc.).
Chemical liver injury is the hepatic injury that is caused by the chemical Hepatoxic substance.These chemical substances comprise chemical toxicant and the some drugs thereof in ethanol, the environment.Liver as the important detoxifcation organ of human body has Hepatic artery and hepatic vein double blood supply.Chemical substance can enter liver by gastrointestinal tract portal vein or body circulation and transform, and liver is subjected to the toxicant infringement in the chemicals easily thus.All exist some to the virose material of liver in the Nature and the human industrial processes, be called " hepatotropic poison ", these poisonous substances are general susceptible in the crowd, incubation period is short; the process of pathological changes is directly related with the dosage of infection, can cause liver hepatic necrosis, fatty distortion, liver cirrhosis and hepatocarcinoma in various degree.Raw material in the commercial production, intermediate product and final raw material all have liver toxicity, therefore need the generation of very vigilant chemical liver poisoming.
The mechanism of chemical toxicant liver injury is important to be had: 1. steatosis, carbon tetrachloride, yellow phosphorus etc. can be disturbed the synthetic and transhipment of lipoprotein, form fatty liver.2. lipid peroxidation, this is the special representing form of toxic liver injury, as carbon tetrachloride in vivo metabolism produce the very strong intermediate product of a kind of oxidability, cause the lipid peroxidation on the biomembrane, destroy the phospholipid of film, change the structure and the function of cell.3. cholestasis reaction, main and liver plasma membrane and microvillus are impaired, cause that the bile acid acatharsia is relevant.
Due to the chemical substance (as ethanol etc.) hepatic injury can by its in hepatocyte metabolism and cause.Enter the ethanol in the hepatocyte, can suppress the mitochondrion tricarboxylic acid cycle by the ratio of rising NADH and NAD, cause intrahepatic fat acid metabolic generation obstacle, oxidation reduction and NADH increase, promote the synthetic of fatty acid, neutral fat is piled up in hepatocyte and steatosis is taken place, serious steatosis can cause hepatic necrosis, proliferation of fibrous tissue in the liver.Liver damage disease, especially fatty liver and abnormal liver function are many to raise with blood lipid level, and the prompting fatty acid metabolism is disorderly to have certain internal relation with fatty liver and hepatic injury.Reported in literature is arranged, and triglyceride (TG) and very low density lipoprotein (VLDL) (VLDL) can promote hepatic stellate cell (HSC) propagation, and be relevant with the generation possibility of hepatic fibrosis.
Fatty acid is closed in many insatiable hungers, ω-3PUFA especially, and the effect of blood lipid regulation level, treatment hyperlipemia and fatty liver be many institutes confirmations.Discover, to oily 8 weeks of rabbit hyperlipidemia model feed sea dog (Canis familiaris L.), can obviously reduce serum total cholesterol and triglyceride, the increasing high density ester gp reduces cholesterol accumulating at liver and heart.Prompting sea dog (Canis familiaris L.) oil has certain preventive and therapeutic effect to cardiovascular disease and fatty liver.
In recent years, many lipid peroxidations that studies confirm that play an important role in the chemical liver injury process.Body endoperoxides increased response can cause that antioxidant ability of organism descends, thereby the generation of free radical is increased.Free radical is the lipid of oxidation cell membrane not only, the more important thing is the protein of oxidation cell membrane, finally causes the infringement of hepatocyte structure and function.Free radical scavenger superoxide dismutase (SOD) and catalase (CAT) etc. can be removed superoxide anion and the hydroxy radical with height reactivity that produces in the lipid peroxidation process respectively, prevent that lipid peroxidation is to histiocytic damage.Experimentation confirms that anti peroxidation of lipid class medicine can have protective effect to hepatic injury.
Summary of the invention
The object of the present invention is to provide the application of a kind of Adeps Phocae vitulinae in preparation control chemical liver injury health product.
Another object of the present invention is to provide the application of a kind of Adeps Phocae vitulinae compositions in preparation control chemical liver injury health product.
Research worker of the present invention confirms that through a large amount of experimental analysiss the Adeps Phocae vitulinae that includes docosahexenoic acid (DHA), eicosapentaenoic acid (EPA) and clupanodonic acid (DPA) can be in the application in the preparation control chemical liver injury health product.
Be preferably, the Adeps Phocae vitulinae compositions of Adeps Phocae vitulinae 99.5% and vitamin E 0.5% composition is as the application of preparation control chemical liver injury health product by weight percentage.
Adeps Phocae vitulinae of the present invention new Arctic Sea dog skin of the hunting fat down that has drawn from, through technologies such as vacuum distillinges, materials such as attached Deproteinization, carbohydrate under cryogenic conditions obtain being the refine Adeps Phocae vitulinae of little yellow, transparency liquid shape.
Refined fur seal of the present invention is by Canadian import, and it is a light yellow oil, has the intrinsic fragrance of Adeps Phocae vitulinae, free from extraneous odour, the visible exogenous impurity of no naked eyes.Physical and chemical index should meet the regulation of table 1.
Table 1 physical and chemical index vitamin
| Project | Method of testing | Index |
| Moisture and volatile matter content, %≤ | GB/T5528 | 0.2 |
| Acid value, mg KOH/g≤ | GB/T5530 | 2.0 |
| Peroxide value, meq/Kg≤ | GB/T5538 | 6.0 |
| Impurity, %≤ | 0.1 |
Wherein, contain docosahexenoic acid 〉=8.22g/100g in the described Adeps Phocae vitulinae, eicosapentaenoic acid 〉=6.09g/100g, clupanodonic acid 〉=3.64g/100g.
The employing of described compositions places capsule forming machine to make soft capsule the mixed liquor of refined fur seal and vitamin E and the edible Gelatinum oxhide of heat fused, and drying is washed the grain sterilization with 75% edible ethanol and formed then.
Vitamin E of the present invention is the edible vitamin E in this area, meets the regulation of GB 14756.
Soft capsule of the present invention is this area pharmagel capsule commonly used, meets the standard of GB6783.
Through great deal of experimental, determine that the capsular technical specification of Adeps Phocae vitulinae of the present invention is as follows:
Table 2 organoleptic indicator
| Project | Index |
| Color and luster | Capsule is little yellow, and content is light yellow to yellow |
| Character | Capsule surface gloss, content are light yellow oil |
| Grow abnormal smells from the patient | Has the intrinsic fragrance of Adeps Phocae vitulinae, free from extraneous odour |
| Impurity | The visible exogenous impurity of no naked eyes |
Table 3 physical and chemical index
| Project | Method of testing | Index |
| Net content, the g/ grain | 0.5 | |
| The net content allowed minus deviation, %≤ | 9.0 | |
| Moisture and volatile matter content, %≤ | GB/T 5528 | 2.0 |
| Disintegration, min≤ | Pharmacopeia regulation in 2000 | 60.0 |
| Acid value, mgKOH/g≤ | GB/T 5530 | 2.0 |
| Peroxide value, meq/kg≤ | GB/T 5538 | 12.0 |
| Vitamin E, mg/100g 〉= | GB/T 12388 | 200 |
| Arsenic (in), mg/kg≤ | GB/T 5009.11 | 1.0 |
| Plumbous (in), mg/kg≤ | GB/T 5009.12 | 1.5 |
| Hydrargyrum (in), mg/kg≤ | GB/T 5009.17 | 0.3 |
Table 4 microbiological indicator
| Project | Method of testing | Index |
| Total plate count, cfu/g≤ | GB 4789.2 | 1000 |
| Coliform, MPN/100g≤ | GB 4789.3 | 40 |
| Mold count, cfu/g≤ | GB 4789.15 | 25 |
| Yeast counts, cfu/g≤ | GB 4789.15 | 25 |
| Pathogenic bacterium | GB 4789.4，GB 4789.5， GB 4789.10，GB 4789.11 | Must not detect |
Annotate: pathogenic bacterium mean Salmonella, shigella, staphylococcus aureus, Hemolytic streptococcus.
The effective component of table 5 Adeps Phocae vitulinae soft capsule
Adeps Phocae vitulinae soft capsule of the present invention, every 500mg recommends consumption to be everyone every day 2 times, each 2.
Function to Adeps Phocae vitulinae of the present invention and Adeps Phocae vitulinae compositions thereof is analyzed, and the generation that fatty acid metabolism and interior free yl effect and lipid peroxide are closed in discovery fatty acid metabolism, especially insatiable hunger has confidential relation.Studies show that EPA and DHA can significantly reduce the content of lipid peroxide malonaldehyde (MDA) in liver, cerebral tissue and the blood, improve the activity of superoxide dismutase (SOD) in liver, the cerebral tissue.Thereby; by activity, removing free radical and the inhibition lipid peroxidation that improves antioxidant in the body; by improving the disorder of liver fat acid metabolic; blood lipid regulation content; reduce cholesterol accumulating and prevent hepatic fibrosis in liver, EPA and DHA should have protective effect to hepar damnification due to the chemical substance.
Therefore, the ω-3PUFA in the Adeps Phocae vitulinae removes free radical, prevents lipid peroxidation to the liver organization cells injury that the reply chemical liver injury does to have protective effect by regulating fatty acid metabolism in blood, the liver.Function test result shows that also taking this product for a long time has auxiliary protection function to chemical liver injury.
Simultaneously vitamin E plays an important role in removing free radical proceed as a kind of antioxidant, can suppress the lipid peroxidation deposits yields effectively, and it is rotten to prevent that insatiable hunger in the product from closing fatty acid oxidation.
Therefore Adeps Phocae vitulinae of the present invention and Adeps Phocae vitulinae compositions thereof have auxiliary protection function to chemical liver injury.
The specific embodiment
Below be embodiments of the invention, the embodiment that the present invention provides is in order to further specify the specific embodiments of the invention scheme, rather than is used for limiting protection scope of the present invention.
Experimental example 1
This experimental example relates to the toxicological assessment test of Adeps Phocae vitulinae of the present invention and Adeps Phocae vitulinae compositions thereof.
One. its mouse oral acute toxicity test of Adeps Phocae vitulinae of the present invention and Adeps Phocae vitulinae compositions thereof, micronucleus test, mouse sperm deformity test, Salmonella reversion test, 30d feeding trial.
Test basis: " food safety toxicological evaluation program and method " GB15193-94
1 material and method
1.1 sample: the Adeps Phocae vitulinae compositions provides faint yellow shape liquid, capsule dress by adding arctic pharmaceutcal corporation, Ltd in growing in vain.Carry out acute toxicity test in mice, PCEMNR micronucleus test, mouse sperm deformity test, Salmonella reversion test, rat 30d feeding trial respectively with its Adeps Phocae vitulinae compositions.
1.2 experimental animal: Kunming kind white mice is provided approval card number: No. [2000] 015, distant real kinoplaszm word by laboratory animal portion of Chinese Medical Sciences University.
The Wistar rat is provided by laboratory animal portion of Chinese Medical Sciences University, approval card number: No. [2000] 015, distant real kinoplaszm word.
1.3 acute toxicity test in mice: select 40 of healthy Kunming kind white mice for use, each 20 of male and female are tested.The mice body weight is 18.0g～22.0g.21.50,10.00,4.64,2.15g/Kg.bw each sex mice is divided into four dosage groups at random, is respectively:, each dosage group is prepared with vegetable oil.Observe 14d behind the mouse stomach continuously.Record poisoning manifestations and death condition.
1.4 genetic toxicity test:
1.4.1Ames test: employing is tested through identifying satisfactory Salmonella typhimurium histidine defect type TA97, TA98, TA100, four test strains of TA102.Adopt the inductive rat liver homogenate of Polychlorinated biphenyls (PCB) as external metabolism activation system.Test establishes 0.313,0.625,1.250,2.500,5 dosage of 5.000mg/ ware, each dosage group is diluted with dimethyl sulfoxide (DMSO), (TA97 of disactivation system, TA98, TA102 are fenaminosulf 50.0 μ g/ wares, and TA100 is folded ammonia sodium 1.5 μ g/ wares to establish blank, solvent control, negative control (distilled water) and positive control simultaneously.Activation system adopts N-2-Fluorenylamine 10.0 μ g/ wares).Adding 0.1ml test strain enrichment liquid, 0.1ml are tried thing solution in top agar, add 0.5ml S9 mixed liquor during metabolism activation, pour into behind the mixing on the bottom culture medium flat plate.Cultivate 48h 37, count every ware and return the change clump count.If being tried the clump count that return to become of thing is to become clump count more than 2 times from beaming back, and has dosage one reaction relation person and then be decided to be the positive.Each dosage do three parallel.A whole set of test repeats under the same conditions to do twice and adds up respectively.
1.4.2 PCEMNR micronucleus test: adopt 24h twice per os administration by gavage in interval to test, irritate stomach amount 0.2ml/10g body weight.With body weight 26.0-29.8g white mice, by the body weight random packet, 10 every group, male and female half and half.With the positive contrast of cyclophosphamide (CP) of 40mg/kg.bw dosage, ripe Oleum Glycines negative control is tried the agent amount and is 0.94,1.88,3.75,7.50g/kg.bw, is assigned to desired concn with ripe Oleum Glycines.6h after the last administration, animal is put to death in the cervical vertebra dislocation, gets femur bone marrow and dilutes smear with calf serum, and methanol is fixed, Giemsa dyeing.Every animal counting 1000 polychromatic erythrocytes (PCE) under optical microscope, microkernel incidence is to contain the PCE permillage of micronucleus.
1.4.3 mouse sperm deformity test: with the sexual maturity male mice of body weight 26.0-30.0g, by the body weight random packet.Tried agent amount 1.88,3.75,7.50g/kg.bw, be assigned to desired concn with ripe Oleum Glycines.With the positive matched group of 40mg/kg.bw cyclophosphamide (CP), the negative matched group of distilled water, ripe Oleum Glycines are solvent control.More than each test group irritate stomach once every day, continuously 5d irritates stomach amount 0.2ml/10g body weight, 30d put to death animal after last was irritated stomach, got the epididymis film-making, Yihong dyeing, several 5 animals of every batch total, the sperm of 1000 structural integrities of every animal counting calculates distortion spermatogenesis rate.
1.530d feeding trial
1.5.1 sample: the Adeps Phocae vitulinae compositions, tried thing with its mouse oral acute toxicity test.
1.5.2 experimental animal: select body weight 55.1-69.5g Wistar rat for use.
1.5.3 test method: rat is divided into matched group and three is at random tried the thing group, it is 3.000g/60kg.bw (press content calculating) that the manufacturer's recommended adult takes in the approved product maximum (body weight is in 60kg) every day.The maximum dose level group is a B group in Coming-of-Age Day 100 times of maximum intake, be 5.000g/kg.bw, below in, low dose group successively decreases with 1/5, be C group 1.000g/kg.bw, 20 times of maximum intake are equivalent to be grown up, D organizes 0.200g/kg.bw, 4 times of the maximum intake that is equivalent to be grown up, every group of 10 rats.B, C, three test group of D are tried thing evenly mix in the normal feedstuff, content is 5.000%, 1.000%, 0.200% according to this.Matched group (A group) feed arm's length basis feedstuff.The rat feed intake is calculated by body weight 10%, feeds separately, and free diet, record rats eating amount, body weight are observed 30d continuously.
1.5.4 observation index:
1.5.4.1 clinical observation: the general performance of animal, behavior, poisoning symptom, and death condition, claim body weight, intake weekly, calculate the food overall utilization.
1.5.4.2 routine blood test and biochemical indicator: get blood in test 31d tail vein, adopt Japanese CA-300 blood cell automatic counter for counting, measure hemoglobin (HGB), erythrocyte (RBC), leukocyte (WBC).Femoral artery is got blood, and the test kit, the Dutch VITALAB-MICRO biochemistry analyzer that adopt Beijing Zhongsheng Biological Engineering High Technology Company to provide are measured glutamate pyruvate transaminase (GPT), glutamic oxaloacetic transaminase, GOT (GOT), blood urea nitrogen (BUN), creatinine (CR), blood glucose (GLU), total protein (TP), albumin (ALB), cholesterol (CHO), triglyceride (TG).
1.5.4.3 pathological anatomy: gross examination of skeletal muscle, organ coefficient, pathological tissue inspection (liver,kidney,spleen, stomach and duodenum)
2. result of the test
2.1 acute toxicity test in mice
Table 6 acute toxicity test in mice result
Manufacturer's recommended adult (body weight is in 60kg) day absorption approved product maximum 0.05/kg.bw, the maximum dose level group is tried thing 21.50/kg.bw, converts to be equivalent to 430 times of Coming-of-Age Day absorption approved product amount.Each dosage group of laboratory observation 14d not poisoning symptom, death toll is zero.Tried the acute toxicity LD of thing to two kinds of sex mices
50All greater than 15.00g/kg.bw according to toxicity grading, belong to nontoxic level.LD
50Greater than 10 times of manufacturer's recommended Coming-of-Age Day intakes, can enter the next stage toxicological experiment.
2.2 genetic toxicity test
2.2.1Ames test:
Table 7Ames result of the test (for the first time)
Table 8Ames result of the test (for the second time)
By table 7,8 as seen, each dosage group is returned change bacterium colony number average and is returned two times that become clump count above negative control group, does not also have dose-response relationship.Illustrate that this is tried thing to Salmonella typhimurium histidine defect type TA97, TA98, TA100, TA102 four bacterial strains, add and do not add S9, all do not present genetoxic.
2.2.2 PCEMNR micronucleus test:
Table 9 mouse bone marrow cells PCE micronucleus test result
Through X 2 test, each dosage group micronuclear rates does not have significance (P＞0.05) with negative group comparing difference, and positive controls and negative control group relatively have highly significant difference (P＜0.01).It is negative that this is tried the PCEMNR micronucleus test.
2.2.3 mouse sperm deformity test:
Table 10 mouse sperm deformity result of the test
Through the Wilconson rank test, each dosage group rate of teratosperm and negative control group comparing difference do not have significance (P＞0.05), and positive controls and negative control group relatively have highly significant (P＜0.01) difference.The mouse sperm deformity test is negative.
2.330d feeding trial
2.3.1 influence to rat body weight:
Table 11 30d feeding trial rat body weight measurement result
By table 11 as seen, each treated animal vegetative activity is normal.Each dosage treated animal body weight and matched group compare, and difference does not have significance (P＞0.05).
2.3.2 the influence to rat total foodstuff utilization rate is as shown in table 12:
30 days feeding trial total foodstuffs of table 12 utilization rate measurement result
By table 12 as seen, tried thing and mixed in the feedstuff feed rat 30 days, the refusing to eat phenomenon does not appear in animal, and each dosage treated animal food utilization and matched group comparing difference do not have significance (P＞0.05).
2.3.3 the hematological examination result is as shown in table 13:
Table 13 feeding trial hematological examination in 30 days result
By table 13 as seen, the hemoglobin of each dosage group (HGB), erythrocyte (RBC), leukocyte (WBC) do not have significance (P＞0.05) with the matched group comparing difference.
2.3.430 the result is as shown in table 14 in it feeding trial biochemical investigation:
Table 1430 day feeding trial biochemical investigation result
By table 14 as seen, the glutamate pyruvate transaminase of each dosage group (GPT), glutamic oxaloacetic transaminase, GOT (GOT), blood urea nitrogen (BUN), creatinine (CR), blood glucose (GLU), total protein (TP), albumin (ALB), cholesterol (CHO), triglyceride (TG) do not have significance (P＞0.05) with the matched group comparing difference.
2.3.5 tried the influence of thing to the dirty body ratio of rat:
The measurement result of 30 days dirty body ratios of feeding trial rat of table 15
By table 15 as seen, each dosage group and matched group comparing difference do not have significance (P＞0.05).
2.3.6 histopathologic examination:
Each experimental group and matched group laboratory animal hair luster, mucosa are ruddy, it is normal, no abnormal to cut open each internal organs position of inspection observation substantially, and when dissected is not also found bladder, common hepatic duct calculus, liver,spleen,kidney color and luster uniform outer surface is smooth, and stomach, duodenum are not also found ulcer and other abnormal change; The liver,spleen,kidney of high dose group and matched group laboratory animal, stomach, duodenum are as follows through the pathological examination result:
Liver: smooth surface, smooth, do not see protuberance and enclosed mass.Finding under the mirror: removing negative control group has the indivedual hepatocyte of a routine buck to have fat to drip existence, other animal livers central veins and hepatic cords and the clear finding of limiting plate, and no abnormality seen changes.One routine hepatocyte fatty of negative control group becomes may belong to the naturality pathological changes, not talkative relevant with this test.
Kidney: smooth surface is smooth, does not see obvious increase and dwindles.Mirror is observed down: the no abnormal change of general structure of kidney.The organizational structure of kidney, each nephron (renal capsule, glomus, proximal tubule, distal tubule etc.) structure is normal, and the renal blood vessels no abnormality seen changes.High dose group and matched group relatively find no significant difference.
Stomach and duodenum: mucous layer, tela submucosa, flesh layer and cast outer mold structure are normal, do not find pathological change.
Spleen: smooth surface is smooth, does not see obvious increase and dwindles.Mirror is observed down: red, white pulp is high-visible, does not see hyperemia and other pathological changes.
The Adeps Phocae vitulinae compositions was mixed in the feedstuff feed rat 30 days, and to each dosage treated animal body weight gain, food utilization, every indexs such as routine blood test, blood biochemistry index, organ coefficient there is no harmful effect.Histopathology is observed, and liver,spleen,kidney, stomach, duodenum there is no significant pathological change.
2.4 conclusion:
Through above a large amount of test result analysis, can think Adeps Phocae vitulinae of the present invention and Adeps Phocae vitulinae compositions thereof through its mouse oral acute toxicity test LD50 greater than 15.00g/kg.bw, belong to nontoxic level according to toxicity grading, micronuclei in mice, mouse sperm deformity are tested, Salmonella reversion test is all negative, the 30d feeding trial is not seen obvious ill-effect.
Experimental example 2
This experimental example relates to Adeps Phocae vitulinae of the present invention and Adeps Phocae vitulinae compositions thereof have auxiliary protection function to chemical liver injury research.
Test basis: " instrument safety toxicological evaluation program and method " GB15193.1-94
1. materials and methods
1.1 sample: Adeps Phocae vitulinae compositions of the present invention is faint yellow oily soft capsule.It is that 3000.0mg/60kg.bw is 50.0mg/kg.bw.d that manufacturer's recommended adult (body weight is in 60kg per capita) takes in the approved product amount.
1.2 animal: the Wistar male white rat, body weight 188.2-212.7 gram provides approval number by animal portion of Chinese Medical Sciences University: distant real animal No. 010.
1.3 reagent and instrument:
Carbon tetrachloride (C CI
4Analytical pure): Beijing Chemical Plant's product batch number: 990401
Glutamate pyruvate transaminase (ALT): Beijing Zhongsheng Biological Engineering High Technology Company produces
Authentication code: (93) are defended the accurate word D-36-3 of medicine number
Glutamic oxaloacetic transaminase, GOT (AST): Beijing Zhongsheng Biological Engineering High Technology Company produces
Authentication code: (91) are defended the accurate word D-20-2 of medicine number
Triglyceride (TG): Beijing Zhongsheng Biological Engineering High Technology Company produces
Authentication code: (95) are defended the accurate word D-17-1 of medicine number
Glutamate pyruvate transaminase, glutamic oxaloacetic transaminase, GOT, triglyceride all adopt Holland to produce the full-automatic analyser of AUTOLABPM4000/ and measure.
1.4 dosage grouping: establish three dosage groups according to 25,5,1 times of human body recommended amounts, be respectively 1250.0mg/kg.bw, 250.0mg/kg.bw, 50.0mg/kg.bw, tried thing and be diluted to desired concn with soybean salad oil, other establishes solvent contrast, negative control group and carbon tetrachloride model group.The solvent contrast gives soybean salad oil, negative control group gives distilled water.
1.5 test method: make the acute chemical hepatic injury model as hepatotoxic agent with 1% carbon tetrachloride, duration of test is tried each dosage group of thing and is irritated Adeps Phocae vitulinae compositions of the present invention (irritating stomach amount 1.0mL/100g.bw) the continuous irrigation stomach 30d that stomach gives various dose, negative control group and carbon tetrachloride hepatic injury model group are irritated stomach and are given distilled water, and the solvent matched group gives soybean salad oil.Put to death animal each treated animal fasting in preceding 24 hours on the 31st day, each dosage group continues to be tried thing, negative control, solvent matched group and model group give distilled water or soybean salad oil respectively, give the Adeps Phocae vitulinae compositions after 1 hour carbon tetrachloride model group and each dosage group irritate stomach and give 1% carbon tetrachloride (irritating stomach amount 0.5mL/100g.bw), give carbon tetrachloride and get blood system from serum after 23 hours, measure the content of TG in ALT, AST value and the hepatic tissue, and get liver and make pathology section, the pathology damage situation of observing liver.Histopathology is observed with hepatocyte " degeneration, necrosis " and is waited as observation index, and quantizes according to every kind of lesion degree, by " (0 minute) ,+(1 minute), ++ (2 minutes), +++(3 minutes), ++ ++ (4 minutes) quantification ".
1.6 experimental data adopts the SPSSforwindow10.0 statistical software to carry out method of analysis of variance.
2. result of the test
2.1 Adeps Phocae vitulinae compositions of the present invention is to the influence of the weight of animals
By table 16 as seen, per os gives the Adeps Phocae vitulinae compositions 30d of the present invention of various dose, and each dosage group, model group, solvent matched group and negative control group compare, and the weight of animals, weightening finish difference do not have significance (P＞0.05).
Table 16 Adeps Phocae vitulinae compositions of the present invention is to the influence of the weight of animals
| Dosage (mg/kg.bw) | Number of animals (only) | Body weight (g) before the test | P | Test back body weight (g) | P | Weightening finish (g) | P |
| Negative control | 10 | 197.0±7.9 | - | 265.6±26.6 | - | 68.6±22.1 | - |
| The solvent contrast | 10 | 197.2±8.6 | 0.970 | 262.0±13.2 | 1.000 | 64.8±10.1 | 1.000 |
| CCI 4 (5ml/kg.bw) | 10 | 196.7±6.9 | 0.926 | 267.0±16.5 | 1.000 | 70.3±12.4 | 1.000 |
| 1250.0 | 10 | 197.0±8.9 | 0.993 | 267.2±15.3 | 1.000 | 70.4±11.4 | 1.000 |
| 250.0 | 10 | 197.5±8.6 | 0.902 | 267.1±11.4 | 1.000 | 69.6±11.5 | 1.000 |
| 50.0 | 10 | 196.3±7.7 | 0.846 | 266.1±13.2 | 1.000 | 69.8±7.6 | 1.000 |
2.2 Adeps Phocae vitulinae compositions of the present invention is to the influence of the TG in animal serum ALT, AST and the hepatic tissue
By table 17 as seen, the ALT of carbon tetrachloride hepatic injury model group, AST, TG are higher than negative control group, and difference has significance (P＜0.05); The ALT of Adeps Phocae vitulinae combination object height of the present invention, middle dosage group is lower than the carbon tetrachloride hepatic injury model group, and difference has significance (P＜0.05); The AST of Adeps Phocae vitulinae compositions high dose group of the present invention is lower than the carbon tetrachloride hepatic injury model group, and difference has significance (P＜0.05).
Table 17 Adeps Phocae vitulinae compositions of the present invention is to the influence among the TG in animal serum ALT AST and the hepatocyte
| Dosage (mg/kg bw) | Number of animals (only) | ALT (U/L) | P | AST (U/L) | P | TG (mg/dL) | P |
| Negative control | 10 | 68±28 | 0.000 ** | 186±69 | 0.002 ** | 170±75 | 0.030 * |
| The solvent contrast | 10 | 67±25 | 0.000 ** | 186±70 | 0.002 ** | 170±75 | 0031 * |
| CCI 4 (5ml/kg.bw) | 10 | 1195±329 | --- | 1115±470 | --- | 450±210 | --- |
| 1250.0 | 10 | 440±328 | 0.001 ** | 466±158 | 0.024 * | 275±128 | 0.456 |
| 250.0 | 10 | 663±373 | 0.049 * | 830±260 | 0.841 | 408±178 | 1.000 |
| 50.0 | 10 | 949±521 | 0.979 | 860±339 | 0.951 | 446±179 | 1.000 |
*p＜0.05
**p＜0.01
2.3 Adeps Phocae vitulinae compositions of the present invention is to the influence of hepatic lesions
By table 18,19 as seen, the liver body ratio of each dosage group of Adeps Phocae vitulinae compositions of the present invention and carbon tetrachloride hepatic injury model group and negative control group comparison animal is not seen notable difference (P＞0.05); Carbon tetrachloride hepatic injury model group (being as the criterion with the change of irritability ball sample, steatosis, endochylema cohesion, four kinds of pathological changes total marks of hepatic necrosis) liver cell lesion degree is heavy than negative control group, and difference has significance (P＜0.01).Adeps Phocae vitulinae compositions high dose group rat hepatocytes lesion degree of the present invention is light than the carbon tetrachloride hepatic injury model group, and difference has significance (P＜0.05).
Table 18 is respectively organized the comparison of rat liver pathological changes
Table 19 is respectively organized rats'liver body ratio and hepatic pathology histological examination result
| Dosage (mg/kg.bw) | Number of animals (only) | Liver weight/body weight (%) | P | Pathological change (respectively organizing average integral) | P |
| Negative control | 10 | 3.63±0.38 | - | 0.10±0.32 | 0.000 ** |
| The solvent contrast | 10 | 3.65±0.42 | 0.914 | 0.00±0.00 | 0.000 ** |
| CCI 4 (5ml/kg.bw) | 10 | 3.79±0.45 | 0.341 | 3.30±1.16 | -- |
| 1250.0 | 10 | 3.57±0.32 | 0.696 | 1.80±0.63 | 0.044 * |
| 250.0 | 10 | 3.61±0.27 | 0.899 | 3.40±0.84 | 0.630 |
| 50.0 | 10 | 3.63±0.36 | 0.976 | 3.20±0.79 | 1.000 |
*p＜0.05
**p＜0.01
2.4 result:
Rat continuous irrigation stomach caused the acute chemical hepatic injury model with carbon tetrachloride on the 31st day after giving 1250.0mg/kg.bw, 250.0mg/kg.bw, 50.0mg/kg.bw Adeps Phocae vitulinae compositions of the present invention 30d.The ALT of carbon tetrachloride hepatic injury model group, AST, TG are higher than negative control group, and difference has significance (P＜0.05); The ALT of Adeps Phocae vitulinae combination object height of the present invention, middle dosage group is lower than the carbon tetrachloride hepatic injury model group, and difference has significance (P＜0.05); The AST of Adeps Phocae vitulinae compositions high dose group of the present invention is lower than the carbon tetrachloride hepatic injury model group, and difference has significance (P＜0.05); Carbon tetrachloride hepatic injury model group (being as the criterion with the change of irritability ball sample, steatosis, endochylema cohesion, four kinds of pathological changes total marks of hepatic necrosis) liver cell lesion degree is heavy than negative control group, and difference has significance (P＜0.01).Adeps Phocae vitulinae compositions high dose group rat hepatocytes lesion degree of the present invention is light than the carbon tetrachloride hepatic injury model group, and difference has significance (P＜0.05).To sum up the result can think that Adeps Phocae vitulinae compositions of the present invention has auxiliary protection function to chemical liver injury.
Experimental example 3
This experimental example relates to the quantitative test of Adeps Phocae vitulinae of the present invention and Adeps Phocae vitulinae compositions thereof, the research of stability test.
Select three batches of Adeps Phocae vitulinae compositionss of the present invention to carry out Detection of Stability, preservation condition: relative humidity 75%, 37 ℃ of temperature, assay method is the same, and the result is as follows:
Table 20 Adeps Phocae vitulinae composition stable testing result
Experimental example 1
The edible Gelatinum oxhide of getting Canadian import refined fur seal 99.5kg, vitamin E 0.5kg mixed liquor and heat fused places capsule forming machine to make soft capsule, and drying is washed the grain sterilization with 75% edible ethanol and formed then.Make 200000 of the capsules of every 0.5g; wherein every 100g contains effective composition: docosahexenoic acid (DHA) 8.35g, eicosapentaenoic acid (EPA) 6.15g, clupanodonic acid (DPA) 3.84g; to the person that has the chemical liver injury oral every day 2 times; each 2; took 90 days, and had the effect that chemical liver injury is had auxiliary protection function.
Claims (2)
1. the application of the Adeps Phocae vitulinae compositions of Adeps Phocae vitulinae 99.5% and vitamin E 0.5% composition in preparation control chemical liver injury health product by weight percentage.
2. the application of Adeps Phocae vitulinae compositions according to claim 1 in preparation control chemical liver injury health product, it is characterized in that: contain docosahexenoic acid 〉=8.22g/100g in the described Adeps Phocae vitulinae, eicosapentaenoic acid 〉=6.09g/100g, clupanodonic acid 〉=3.64g/100g.
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| CN102551041A (en) * | 2010-12-08 | 2012-07-11 | 浙江金诺康生物制药有限公司 | Refined seal oil, seal oil soft capsules and application of seal oil soft capsules in preparing health products for improving chemical liver injury |
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| CN1410074A (en) * | 2001-09-30 | 2003-04-16 | 中国药品生物制品检定所 | Application of fur seal oil in preparation of medicine for treating acute liver damage |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1410074A (en) * | 2001-09-30 | 2003-04-16 | 中国药品生物制品检定所 | Application of fur seal oil in preparation of medicine for treating acute liver damage |
Non-Patent Citations (2)
| Title |
|---|
| 维生素E在肝损伤防治中的地位和作用. 王雅凡等.《国外医学》流行病学传染病学分册，第22卷第5期. 1995 |
| 维生素E在肝损伤防治中的地位和作用. 王雅凡等.《国外医学》流行病学传染病学分册，第22卷第5期. 1995 * |
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