CN103565847A - Preparation method of raja porosa extract as well as application of raja porosa extract in alcoholic liver protection medicament - Google Patents
Preparation method of raja porosa extract as well as application of raja porosa extract in alcoholic liver protection medicament Download PDFInfo
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- CN103565847A CN103565847A CN201310513265.7A CN201310513265A CN103565847A CN 103565847 A CN103565847 A CN 103565847A CN 201310513265 A CN201310513265 A CN 201310513265A CN 103565847 A CN103565847 A CN 103565847A
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- 241000039233 Okamejei kenojei Species 0.000 title claims abstract description 47
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- 210000004185 liver Anatomy 0.000 title claims abstract description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 36
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- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 8
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- 241000251468 Actinopterygii Species 0.000 claims description 5
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- 238000001556 precipitation Methods 0.000 claims description 3
- 239000002689 soil Substances 0.000 claims description 3
- 238000010025 steaming Methods 0.000 claims description 3
- 238000005550 wet granulation Methods 0.000 claims description 2
- 206010067125 Liver injury Diseases 0.000 abstract description 10
- 231100000753 hepatic injury Toxicity 0.000 abstract description 10
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- 102000004190 Enzymes Human genes 0.000 abstract 1
- 108090000790 Enzymes Proteins 0.000 abstract 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 abstract 1
- 239000003963 antioxidant agent Substances 0.000 abstract 1
- 230000003078 antioxidant effect Effects 0.000 abstract 1
- 238000009835 boiling Methods 0.000 abstract 1
- 238000004140 cleaning Methods 0.000 abstract 1
- 238000001035 drying Methods 0.000 abstract 1
- 239000000413 hydrolysate Substances 0.000 abstract 1
- 235000013372 meat Nutrition 0.000 abstract 1
- 230000003617 peroxidasic effect Effects 0.000 abstract 1
- 230000001376 precipitating effect Effects 0.000 abstract 1
- 239000011814 protection agent Substances 0.000 abstract 1
- 238000000967 suction filtration Methods 0.000 abstract 1
- 238000005406 washing Methods 0.000 abstract 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 19
- 102000019197 Superoxide Dismutase Human genes 0.000 description 16
- 108010012715 Superoxide dismutase Proteins 0.000 description 16
- 238000012360 testing method Methods 0.000 description 9
- 208000007848 Alcoholism Diseases 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 241000700159 Rattus Species 0.000 description 5
- 201000007930 alcohol dependence Diseases 0.000 description 5
- 238000003304 gavage Methods 0.000 description 5
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 4
- 238000010171 animal model Methods 0.000 description 4
- 230000037396 body weight Effects 0.000 description 4
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 4
- 230000000630 rising effect Effects 0.000 description 4
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- 238000011160 research Methods 0.000 description 3
- 238000012449 Kunming mouse Methods 0.000 description 2
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- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- 229960003180 glutathione Drugs 0.000 description 2
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- 230000002440 hepatic effect Effects 0.000 description 2
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- 235000019359 magnesium stearate Nutrition 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
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- 238000010998 test method Methods 0.000 description 2
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- 241000119937 Actinotrichia Species 0.000 description 1
- 208000007082 Alcoholic Fatty Liver Diseases 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 206010009208 Cirrhosis alcoholic Diseases 0.000 description 1
- 206010012335 Dependence Diseases 0.000 description 1
- 206010016262 Fatty liver alcoholic Diseases 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 206010019728 Hepatitis alcoholic Diseases 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 208000005374 Poisoning Diseases 0.000 description 1
- 241001415775 Rajidae Species 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
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- 238000011047 acute toxicity test Methods 0.000 description 1
- 208000026594 alcoholic fatty liver disease Diseases 0.000 description 1
- 208000002353 alcoholic hepatitis Diseases 0.000 description 1
- 208000010002 alcoholic liver cirrhosis Diseases 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
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- 239000008103 glucose Substances 0.000 description 1
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- 229940041476 lactose 100 mg Drugs 0.000 description 1
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- 238000011017 operating method Methods 0.000 description 1
- 229960001412 pentobarbital Drugs 0.000 description 1
- WEXRUCMBJFQVBZ-UHFFFAOYSA-N pentobarbital Chemical compound CCCC(C)C1(CC)C(=O)NC(=O)NC1=O WEXRUCMBJFQVBZ-UHFFFAOYSA-N 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a preparation method of a raja porosa extract, and an application of the raja porosa extract in preparing an alcoholic liver protection medicament. The preparation method is characterized by comprising the following steps: boiling fresh raja porosa, subsequently peeling off the meat, washing the soft bone in clean water, crushing the soft bone, adding a NaOH solution according to a material-liquid mass ratio of 1:(1-1.5), stirring and extracting for 4 hours, subsequently adjusting the pH value to be 6 to 7 by using HCl, filtering, adding NaOH into the filtrate to adjust the PH value to be 8.5-9.0, adding compound protease to hydrolyze for 4 hours according to a mass ratio of the material liquid to the compound protease of 1,000:1, heating the hydrolysate for 30 minutes to be 80 DEG C, adding HCl to adjust the pH value to be 2, standing, performing suction filtration on the solution at the upper layer by using kieselguhr, adding NaOH to adjust the pH value to be 10.5, decoloring by using hydrogen peroxide, subsequently filtering, adding HCl into the filtrate to adjust the pH value to be 7, carrying out reduced-pressure concentration on the filtrate to be a small volume, precipitating and drying so as to obtain the raja porosa extract. The raja porosa extract can be used as an alcoholic liver injury protection agent as the activity of an antioxidant enzyme is improved and the cleaning capability of an organism to a peroxidatic reaction product is improved.
Description
Technical field
The present invention relates to a kind of preparation method of Raja porosa extract and in preparation, alcoholic liver injury is protected to the application in medicine.
Background technology
Alcoholism and addiction have become global problem at present, and long-term heavy drinking can cause the infringement of the many organs of whole body, more obvious to the infringement of liver especially.Chronic alcoholism can cause the symptoms such as alcoholic fatty liver, alcoholic hepatitis, alcoholic cirrhosis, fibrosis, and canceration can occur severe patient.At present; for the poisoning treatment of alcoholic liver, adopt comprises aminoacid and glucose infusion, multivitamin more; maintain water, electrolyte and acid-base balance etc., many research worker all try hard to find a kind of can effectively prevention with the liver protecting to alleviate the medicine of ethanol to hepar damnification effect in recent years.
Raja porosa (Raja porosa G ü nther) is commonly called as labor sole, Lao Zi, Pu fish etc., belong to Rajidae, in Huang, the Bohai Sea, all have throughout the year, output is large, for object is fished in important fishery, its Endoskeleton is cartilage entirely, and fin ray is actinotrichia, and the research and development of Raja porosa extract are had to certain science and using value.To the preparation method of Raja porosa extract and the applied research in preparing alcoholic liver injury protection medicine thereof so far there are no report.
Summary of the invention
The object of this invention is to provide a kind of preparation method and the application in preparing alcoholic liver injury protection medicine thereof of Raja porosa extract, to meet the demand of prior art.A kind of preparation method of Raja porosa extract, it is characterized in that after fresh Raja porosa steaming and decocting, rejecting the flesh of fish, cartilage is rinsed, rubbed with clear water, by feed liquid mass ratio, be that 1:1.5 adds NaOH solution, stir and extract 4 h, then with HCl, regulate pH=6～7, filter, in filtrate, add NaOH and regulate pH=8.5～9.0, by the mass ratio of feed liquid and compound protease 1000:1, add composite protease hydrolysis 4 h.By 80 ℃ of heating 30 min of hydrolyzed solution, add HCl to adjust pH=3, standing, Pumex soil sucking filtration for upper solution, adds NaOH to adjust pH=10.5, after hydrogen peroxide decolouring, filters, filtrate adds HCl to adjust pH=7, and filtrate decompression is concentrated into small size, after precipitation is dry, obtains Raja porosa extract.
The application of above-mentioned Raja porosa extract in preparing alcoholic liver injury protection medicine.
The present invention adopts the effect to glutathion peroxidase (GSH-PX), superoxide dismutase (SOD) and malonaldehyde (MDA) of Raja porosa extract that alcohol-induced liver injury in rats animal model tested preparation.Experiment shows; Raja porosa extract of the present invention can make SOD, GSH-PX content raise; MDA content obviously reduces; by strengthening the activity of antioxidase; the removing ability of enhancing body to peroxidation reaction product; thereby performance is to alcoholic liver injury protective effect, and therefore Raja porosa extract of the present invention can be used as alcoholic liver injury protective agent.
the specific embodiment
Below by embodiment and test example, further set forth the preparation of Raja porosa extract of the present invention, with and at the beneficial effect to alcoholic liver injury protection application.
Embodiment
The first step: the preparation of Raja porosa extract
The flesh of fish will be rejected after fresh Raja porosa steaming and decocting, cartilage is rinsed, rubbed with clear water, by feed liquid mass ratio, be to add NaOH solution at 1: 1.5, stir and extract 4 h, so with HCl, regulate pH=6～7, filter, in filtrate, add NaOH and regulate pH=8.5～9.0, by the mass ratio of feed liquid and compound protease 1000:1, add composite protease hydrolysis 4 h.By 80 ℃ of heating 30 min of hydrolyzed solution, add HCl to adjust pH=3, standing, Pumex soil sucking filtration for upper solution, adds NaOH to adjust pH=10.5, after hydrogen peroxide decolouring, filters, filtrate adds HCl to adjust pH=7, and filtrate decompression is concentrated into small size, after precipitation is dry, obtains Raja porosa extract.
Second step: the preparation of preparation
(1) preparation of Raja porosa extract capsule preparation of the present invention
Take respectively Raja porosa extract 100mg, starch 395mg, magnesium stearate 5mg prepared by above-mentioned steps, evenly mix, be filled to capsule.
(2) preparation of Raja porosa extract tablet of the present invention
Take respectively Raja porosa extract 100mg, starch 295mg, lactose 100mg prepared by above-mentioned steps, evenly mix, with wet granulation, dry, after granulate, mix with 5mg magnesium stearate, be pressed into tablet.
Test example 1: the pharmacodynamic study of Raja porosa extract for treating alcoholic liver injury effect
1, test material
(1) Raja porosa
(2) superoxide dismutase (SOD) test kit, glutathion peroxidase (GSH-PX) test kit and malonaldehyde (MDA) test kit all build up Bioengineering Research Institute purchased from Nanjing
(3) male Wistar rat, the laboratory animal department of the Chinese Academy of Sciences of Tongji Medical College, Huazhong Science and Technology Univ. provides, the animal quality certification number: SCXK (Hubei Province) 2004-0007
2, test method:
40 of male Wistar rats, are divided into 4 groups at random by body weight, and A group is blank group, and B group is ethanol model group, and C, D group are respectively that Raja porosa extract of the present invention is low, high dose intervention group, give respectively 50,100 mgkg
-1d
-1gavage, except blank group, all the other laboratory animals give 50 % ethanol 8 mlkg
-1d
-1gavage, continues 2 weeks; The 3rd week is 12 mlkg ethanol dosage escalation
-1d
-1, continuing gavage 6 weeks, after last gavage, water 12h is can't help in fasting, gives sodium pentobarbital anesthesia, abdominal aortic blood, separation of serum takes out hepatic tissue on ice platform, and the operating procedure providing according to test kit is measured SOD, GSH-PX and MDA.
3, experimental result
(1) Raja porosa extract of the present invention is to GSH-PX, SOD in alcoholism rat blood serum and MDA content influence
With the comparison of blank group, in ethanol model group serum, GSH-PX, SOD content reduce the rising of MDA content; With the comparison of ethanol model group, Raja porosa extract of the present invention is low, high dose group all can suppress the reduction of GSH-PX, SOD and the rising of MDA content.
Table 1 Raja porosa extract of the present invention is to GSH-PX, SOD in alcoholism rat blood serum and MDA content influence
| Group | GSH-PX(umol·L -1) | SOD(U·ml -1) | MDA (nmol·ml -1) |
| Blank group | 2263.2±27.0 | 263.4±4.36 | 162.4±12.4 |
| Essence model group | 1630.2±25.3 | 202.8±10.2 | 203.1.0±15.1 |
| Fish extract is low | 1831.0±14.3? | 216.5±22.0 | 181.1±14.8 |
| High | 1926.5±17.3 | 226.3±12.3 | 195.0±20.3 |
(2) Raja porosa extract of the present invention is to GSH-PX, SOD, MDA content influence in alcoholism liver tissues of rats
With the comparison of blank group, in ethanol model group hepatic tissue, GSH-PX, SOD content reduce the rising of MDA content; With the comparison of ethanol model group, Raja porosa extract of the present invention is low, high dose group all can suppress the reduction of GSH-PX, SOD and the rising of MDA content.
Table 2 Raja porosa extract of the present invention is to GSH-PX, SOD, MDA content influence in alcoholism liver tissues of rats
| Group | GSH-PX(umol·L -1) | SOD(U·ml -1) | MDA (nmol·ml -1) |
| Blank group | 1060.7±22.1 | 293.8±27.9 | 18.6±6.5 |
| Ethanol model group | 742.8±30.7 | 179.1±16.7 | 28.5±5.3 |
| Ray fish extract is low | 780.1±27.5 | 198.8±26.3 | 25.7±5.6 |
| High | 806.7±36.6 | 213.4±17.9 | 22.5±3.4 |
Test example 2: the acute toxicity test of Raja porosa extract of the present invention
1, experimental animal:
Kunming mouse, body weight 20 ±2g，You Qingdao City medicine inspecting institute Experimental Animal Center provide
2, medicine:
Raja porosa extract of the present invention
3, test method:
According to National Drug Administration, promulgate " the study of tcm new drug specification requirement " of revision, Raja porosa extract of the present invention is carried out to acute toxicity testing.Because being subject to drug level and volume restrictions, cannot measure LD
50therefore, carry out maximum dosage-feeding experiment.Get 30 of Kunming mouses, female half and half, ig, gives capsule Chinese medicine 1g/mL, every 0.8 mL, i.e. and the maximum gavage volume of mice, with observation post administration mice active state, diet, feces, breathing, body weight and death condition, Continuous Observation 14d.
4, result of the test:
Each organizes well-grown after mouse stomach administration, and body weight increases, and behavior is normal, is showed no death and significant reaction.By after sacrifice of animal, each main organs of perusal is without abnormal phenomena.
Result shows, Raja porosa extract maximal dose oral application of the present invention without obvious damage, is not found the toxic reaction to body generation to animal, is a kind of safe and reliable medicine.
Claims (3)
1. the preparation method of a Raja porosa extract, it is characterized in that after fresh Raja porosa steaming and decocting, rejecting the flesh of fish, cartilage is rinsed with clear water, rub, by feed liquid mass ratio, be to add NaOH solution at 1: 1.5, stir and extract 4 h, then with HCl, regulate pH=6～7, filter, in filtrate, add NaOH and regulate pH=8.5～9.0, by the mass ratio of feed liquid and compound protease 1000:1, add composite protease hydrolysis 4 h, by 80 ℃ of heating 30 min of hydrolyzed solution, add HCl to adjust pH=3, standing, Pumex soil sucking filtration for upper solution, add NaOH to adjust pH=10.5, after hydrogen peroxide decolouring, filter, filtrate adds HCl to adjust pH=7, filtrate decompression is concentrated into small size, after precipitation is dry, obtain Raja porosa extract.
2. the preparation method of Raja porosa extract as claimed in claim 1, also comprise: extract is made to the capsule of this extract 100mg or 200mg specification by conventional pharmacy means, or made the tablet containing this extract 100mg or 200mg specification by wet granulation technology.
3. the application of the Raja porosa extract described in claim 1 in preparing alcoholic liver protection medicine.
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| CN201310513265.7A CN103565847B (en) | 2013-10-28 | 2013-10-28 | The preparation method of Raja porosa extract and the application in alcoholic liver protection medicine |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310513265.7A CN103565847B (en) | 2013-10-28 | 2013-10-28 | The preparation method of Raja porosa extract and the application in alcoholic liver protection medicine |
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| CN103565847B CN103565847B (en) | 2015-09-02 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105125584A (en) * | 2015-09-22 | 2015-12-09 | 青岛大学 | Preparation method of Chimaera phantasma Jordan et Snyder extract and application of Chimaera phantasma Jordan et Snyder extract to preparation of alcoholic liver injury protective drug |
| CN105193851A (en) * | 2015-09-22 | 2015-12-30 | 青岛大学 | Application of raja porosa extract in alcoholic brain damage protection medicine |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101913583A (en) * | 2010-08-20 | 2010-12-15 | 曹荣军 | Method for extracting chondroitin, collagen and high-calcium powder from animal cartilage |
| CN103114116A (en) * | 2013-01-24 | 2013-05-22 | 林丽艳 | Method for producing chondroitin sulfate and coproducing peptone, collagen peptide and cartilage calcium powder |
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2013
- 2013-10-28 CN CN201310513265.7A patent/CN103565847B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101913583A (en) * | 2010-08-20 | 2010-12-15 | 曹荣军 | Method for extracting chondroitin, collagen and high-calcium powder from animal cartilage |
| CN103114116A (en) * | 2013-01-24 | 2013-05-22 | 林丽艳 | Method for producing chondroitin sulfate and coproducing peptone, collagen peptide and cartilage calcium powder |
Non-Patent Citations (1)
| Title |
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| 刘宪锋: "硫酸软骨素双酶法生产新工艺", 《中国药业》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105125584A (en) * | 2015-09-22 | 2015-12-09 | 青岛大学 | Preparation method of Chimaera phantasma Jordan et Snyder extract and application of Chimaera phantasma Jordan et Snyder extract to preparation of alcoholic liver injury protective drug |
| CN105193851A (en) * | 2015-09-22 | 2015-12-30 | 青岛大学 | Application of raja porosa extract in alcoholic brain damage protection medicine |
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