CN1250230C - Application of fur seal oil in preparation of medicine for treating fatty liver - Google Patents
Application of fur seal oil in preparation of medicine for treating fatty liver Download PDFInfo
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- CN1250230C CN1250230C CNB011355174A CN01135517A CN1250230C CN 1250230 C CN1250230 C CN 1250230C CN B011355174 A CNB011355174 A CN B011355174A CN 01135517 A CN01135517 A CN 01135517A CN 1250230 C CN1250230 C CN 1250230C
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- fatty liver
- adeps phocae
- phocae vitulinae
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Abstract
The present invention relates to an application of seal's fat for preparing medicine for treating fatty liver. Researchers find that when the seal's fat of the present invention is continuously poured and administrated to stomachs at the dose of 4.3 to 10 ml/kg, the increase of the content of mouse triglyceride caused by DL-ethionine can be obviously inhibited to mouse fatty liver caused by DL-ethionine. Therefore, under the situation that the seal's fat is used as medicine, the seal's fat has obvious effect on treating fatty liver.
Description
Technical field
What the present invention relates to is the application of Adeps Phocae vitulinae as the preparation medicine for treating fatty liver.
Background technology
As everyone knows, patient's quantity of diseases such as diabetes, hepatopathy, prostate, hyperlipidemia is very huge, and in the practical application, has special positive effect and the medicine of almost being free from side effects is difficult to confirm.
It has been generally acknowledged that patient's quantity that Eskimos and Japanese coastal fisherman suffer from this type of disease is significantly lower than general crowd, research worker believe this phenomenon and eat the fur seal meat that is rich in unsaturated fatty acid with them, oil is relevant.
But, do not have concrete zoopery or clinical effectiveness to be proved so far.
Described in the present invention fur seal typically refers to a kind of mammal of life Newfoundland, North Atlantic Region, the arctic, in their body fats unsaturated fatty acid content up to 25%, the intravital fatty acid molecule structural similarity of its molecular structure and people.
Adeps Phocae vitulinae of the present invention (claiming seal oil again) is meant that the subcutaneous fat by fur seal refines the all-natural product that forms.
Result of study shows, containing in the body of a large amount of needed by human in the Adeps Phocae vitulinae can not synthetic voluntarily ω-3 be long carbochain polyene unsaturated fatty acid and a spot of Squalene (SPUALENE).
Summary of the invention
The Adeps Phocae vitulinae that pharmacological experiment of the present invention adopts is to extract the little yellow oily liquid that forms in the fur seal body, provide in soft capsule (0.5g/ grain) mode by Canadian Atlantic Ocean wholesome food company limited, lot number is 11656, and described Adeps Phocae vitulinae adopts room temperature preservation at laboratory.
For the further composition and the content of clear and definite above-mentioned Adeps Phocae vitulinae, below the composition of Adeps Phocae vitulinae used in the present invention is analyzed and confirmed.
Prior art shows that the ω that extracts-3 is that the chemical main composition of polyene unsaturated fatty acid is EPA, DHA and DPA from Adeps Phocae vitulinae, wherein:
EPA is meant eicosapentaenoic acid;
DPA is meant clupanodonic acid;
DHA is meant docosahexenoic acid;
Research worker of the present invention is carried out the mensuration of projects such as ultra-violet absorption spectrum, infrared absorption spectroscopy, gas chromatogram, gas chromatography-mass spectrometry chromatogram to the Adeps Phocae vitulinae of above-mentioned acquisition, and the chemical constitution of described sample is confirmed, and is specific as follows:
The Adeps Phocae vitulinae test result of samples that the present invention adopts is:
(1) ultra-violet absorption spectrum
With chloroform (analytical pure) sample is made into and carries out the long UV scanning of all-wave behind the 5.9mg/ml solution, the model of employed instrument is U-3210spectrophotometer (Japan manufacturing); Referring to accompanying drawing 1, its result shows that sample has absorption maximum at the 249.8nm place, can prove and contain a large amount of unsaturated bonds in the sample;
(2) infrared absorption spectroscopy
Referring to accompanying drawing 2, its result shows, at 3010cm
-1And 1745cm
-1The strong absorption at place be respectively unsaturated=C-H and＞characteristic absorption of C=O stretching vibration; 1655cm
-1The weak absorption at place meets the stretching vibration characteristic absorption (1665-1635cm of cis-double bonds (R1-C=C-R2)
-1, trans is 1675-1665cm
-1), H/ H722cm
-1The absworption peak at place meets the characteristic absorption (730-665cm of cis-double bonds=C-H bending vibration
-1, trans is 980-960cm
-1).
Comprehensive infrared and ultraviolet spectral analysis can be thought to be mainly the cis non-conjugated fatty acid in the sample.
(3) gas chromatogram
In this detection, the accepted standard product are provided by SIGMA company, are respectively:
Cis-5,8,11,14,17-EICOSAPENTAENOIC ACID METHYL ESTER, methyl eicosapentaenoic acid;
Cis-4,7,10,13,16,19-DOCOSAHEXAENOIC ACID ETHYL ESTER, docosahexaenoic acid ethyl;
Cis-7,10,13,16,19-DOCOSAPENTAENOIC ACID METHYL ESTER, clupanodonic acid methyl ester;
Sample is the Adeps Phocae vitulinae of above-mentioned acquisition, analyzing fatty acid with the GC method need be with its ethyl esterization in order to gasification, specifically be to get described sample 80 μ l, the NaOH-ethanol liquid 1ml that adds 0.5ml/l, fill N2, jump a queue, jolting adds boron trifluoride ethanol liquid 1.5ml to little oil droplet complete obiteration in 50 degrees centigrade of water-baths.Mix homogeneously is put 5min in 50 degree water-baths, take out cooling, and adding normal heptane 1ml, saturated NaCl is the 2ml mix homogeneously, layering.Get upper strata liquid in another test tube, add a small amount of anhydrous NA2SO4, fill N2, place in 4 degree and treat that GC analyzes.
Employed instrument is HP6890;
GC conditions: glass filled post 2m * 3mm, 10%DEGS pickling, 101 white carriers, carrier gas N2, flow velocity 25ml/min, detector FID, column temperature 180 degree, vaporizer 210 degree, detector 270 degree, sample size 1ul.
With standard substance normal hexane standardize solution, be configured to 2.5mg/ml solution, precision is measured 1ul and is carried out the gas phase assay, and same method is measured sample, and the result is that the retention time of sample and reference substance is consistent.
Can be referring to accompanying drawing 3-7.
(4) gas-matter coupling method
The instrument that adopts is a U.S. Trace 2000GC/Trace MS type gas chromatograph-mass spectrometer, and all reagent are analytical pure.
Experiment condition is:
GC conditions: the PEG-20M glass capillary column (2m * 0.32mm); Column temperature 40 degree keep 1min, are warming up to 200 degree with 10 degree/min, keep 5min, are warming up to 210 degree with 30 degree/min again, keep 2min; Press 3.0kpa before the carrier gas N2, post, split ratio 1: 30, injector temperature 250 degree; Ion source temperature 250 degree, ionization voltage 70Ev, mass scanning scope 350AmU.S-1; Sample size 1ml.
Through mass spectrogram analysis, determine that the molecular weight of sample and standard substance conforms to.
So far, molecular weight, composition, the content and structure of of the Adeps Phocae vitulinae that the present invention is adopted have obtained confirming.
In order to confirm that Adeps Phocae vitulinae of the present invention causes curative effect aspect the large and small Mus fatty liver at treatment DL-ethionine, carried out following animal experiment, experiment purpose is to understand described Adeps Phocae vitulinae to cause pharmacotoxicological effect and effect aspect the large and small Mus fatty liver at the described DL-ethionine of treatment, and concrete method and result are as follows:
Experimental example one
The purpose of this experimental example is to understand described Adeps Phocae vitulinae to cause pharmacotoxicological effect and effect aspect the mice fatty liver at the described DL-ethionine of treatment, and concrete method and result are as follows:
Adeps Phocae vitulinae is to extract the little yellow oily liquid that forms in the fur seal body, is produced this laboratory room temperature preservation by Atlantic Ocean wholesome food company limited.Press the 20g body weight during test and irritate stomach 0.6ml, height, in, low dosage all uses dilute liquid medicine.
Positive control drug is a bifendate, every ball content 1.5mg, and lot number 990527, the accurate word (1996) of medicine is defended No. 121020 in the authentication code capital, is produced by Beijing synergism pharmaceutical factory, purchases in pharmacy of Beijing synergism pharmaceutical factory.Before the experiment pill is crushed in mortar, add distilled water while grinding, make the suspendible medicinal liquid of 8mg/ml, the mouse stomach volume is pressed the 0.5ml/20g body weight.
Model drug is the DL-ethionine, powder, and lot number 98H5018 is produced by SIGMA company, is bought on behalf by supply and marketing section of this institute.Take by weighing the DL-ethionine before the experiment on 1/0,000 balances, make the suspendible medicinal liquid of 10mg/ml, the mouse stomach volume is pressed the 0.5ml/20g body weight.
Every of blank group mice is irritated stomach 0.5ml distilled water.
Biochemical reagents and other reagent, triglyceride test kit are Beijing northization fine chemicals company limited production, and normal saline, olive oil are purchased in Beijing pharmacy.
Experimental animal is a cleaning level Kunming mouse, mice body weight 19-21g, and the animal quality quality certification number: the capital is moving is betrothed to (2000) No. 012, is provided by the Nat'l Pharmaceutical ﹠ Biological Products Control Institute experimental center.Observed two days before the experiment, choosing is healthy, and active animal is used for test.
Get 60 of mices, male and female half and half, body weight 19-21g is divided into 6 groups at random: i.e. blank group, DL-ethionine model 0.25g/kg group, positive control drug bifendate 0.2g/kg group, Adeps Phocae vitulinae 10ml/kg group, 5ml/kg, 2.5ml/kg group, 10 every group.Blank group and model control group are irritated the stomach feedwater every day, three dosage groups and positive control drug group gastric infusion every day totally 9 days, at the 7th day, except that the blank group, all add the DL-ethionine in addition and once irritate stomach, in the 9th day broken end sacrificed by exsanguination animal, get liver and blot bloodstain and claim its weight with filter paper, with normal saline by making liver homogenate, centrifugal 5 minutes of 2500rpm at 1: 5, get supernatant and measure content of triglyceride by the test kit requirement, the mensuration wavelength is 550nm.Measure the significance of respectively organizing content of triglyceride difference with two groups of t checks.
Described result sees table 1 for details; Mice is in high spirits during the administration, and activity is not seen drug-induced untoward reaction freely.
Table 1 Adeps Phocae vitulinae is to the influence () of triglyceride
| Group | Dosage ml/kg | Number of animals n | x±SD.mg/g | The p value |
| Blank model contrast bifendate Adeps Phocae vitulinae | —— 0.25g 0.2g 2.5 5 10 | 10 10 10 10 10 10 | 1.61±0.65 4.88±2.13* 2.43±1.42 5.12±4.99 2.25±1.13 1.91±0.68 | —— P＜0.05 p＞0.05 p＜0.05 p＜0.01 |
Compare with the blank group
*P＜0.01
Adeps Phocae vitulinae is to the influence (two) of triglyceride
| Group | Dosage ml/kg | Number of animals n | x±SD.mg/g | The p value |
| Blank model contrast bifendate Adeps Phocae vitulinae | —— 0.25g 0.2 |
10 10 10 10 10 10 | 4.96±0.87 14.01±3.61* 7.06±0.85 6.27±1.34 7.20±0.86 6.13±1.46 | —— P＜0.01 p＜0.01 p＜0.01 p＜0.01 |
Compare with the blank group
*P＜0.01
The result shows that the content of triglyceride of blank group is in range of normal value; The numerical value of model control group raises then very obvious; And the Adeps Phocae vitulinae 10ml/kg of positive controls, the numerical value of 5ml/kg group all has obvious reduction, has significant differences and significant difference (p＜0.01, p＜0.05) with the analysis of model control group comparative statistics.
Conclusion
Adeps Phocae vitulinae is under 5ml/kg-10ml/kg dosage, continuously gastric infusion is 9 days, and the fatty liver to due to the mice DL-ethyl ammonia sulfate can significantly reduce the raising of content of triglyceride, can effectively prevent the generation of fatty liver, card has shown that Adeps Phocae vitulinae has lipotropic preferably effect.
To achieve these goals, the technical solution used in the present invention is: the model that causes the mice fatty liver with the DL-ethionine, to Adeps Phocae vitulinae under 2.5ml/kg-10ml/kg dosage, 9 days pharmacodynamic action of continuous irrigation stomach is observed, and understands Adeps Phocae vitulinae causes mice fatty liver model to the DL-ethionine pharmacotoxicological effect.Found that: this medicine can obviously suppress the DL-ethionine and cause increasing of mice triglyceride content.
Experimental example two
The purpose of this experimental example is to understand described Adeps Phocae vitulinae to cause pharmacotoxicological effect and effect aspect the rat fat liver at the described DL-ethionine of treatment, and concrete method and result are as follows:
Test material
Adeps Phocae vitulinae is to extract the little yellow oily liquid that forms in the fur seal body, is produced this laboratory room temperature preservation by Atlantic Ocean wholesome food company limited.Press the 140g body weight during test and irritate stomach 1.2ml (high dose use stock solution) or 1ml (in, low dosage uses dilute liquid medicine).
Positive control drug is a bifendate, and every ball contains measures 1.5mg, lot number 990527, and the accurate word (1996) of medicine is defended No. 121020 in the authentication code capital, is produced by Beijing synergism pharmaceutical factory, purchases in pharmacy of Beijing synergism pharmaceutical factory.Before the experiment pill is crushed in mortar, add distilled water while grinding, make the concrete suspension of 22.5mg/ml, the rat oral gavage volume is pressed the 1ml/140g body weight.
Model drug is the DL-ethionine, powder, and lot number 98H5018 is produced by SIGM company, is bought on behalf by supply and marketing section of this institute.Take by weighing the DL-ethionine before the experiment on 1/0,000 balances, make the suspension of 28mg/ml, the rat oral gavage volume is pressed the 1ml/140g body weight.
Every of blank group rat is irritated stomach 1ml distilled water.
Biochemical reagents and other reagent, triglyceride test kit are Beijing northization fine chemicals company limited production, and normal saline, olive oil are purchased in Beijing pharmacy.
Experimental animal is a cleaning level wister rat, rat body weight 130g-150g, and the animal quality quality certification number: the capital is moving is betrothed to (2000) No. 012, is provided by the Nat'l Pharmaceutical ﹠ Biological Products Control Institute experimental center.Observed two days before the experiment, choosing is healthy, and active animal is used for test.
Method and result
Be taken at 48 of Mus, male and female half and half, body weight 130-150g is divided into 6 groups at random: promptly blank group, DL-ethionine model 0.2g/kg group, positive control drug bifendate 0.1g/kg group, Adeps Phocae vitulinae 8.6ml/kg group, 4.3ml/kg, 2.2ml/kg group, 8 every group.Blank group and model control group are irritated the stomach feedwater every day, three dosage groups and positive controls gastric infusion every day totally 9 days, at the 7th day, except that the blank group, all add the DL-ethionine in addition and once irritate stomach, in the 9th day broken end sacrificed by exsanguination animal, get liver and blot bloodstain and claim its weight with filter paper, with normal saline by making liver homogenate, centrifugal 5 minutes of 2500rpm at 1: 5, get supernatant and measure content of triglyceride by the test kit requirement, the mensuration wavelength is 550nm.Measure the significance of respectively organizing content of triglyceride difference with two groups of t checks.The result sees table 2 for details.Rat is in high spirits during the administration, and activity is not seen drug-induced untoward reaction freely.
Table 2 Adeps Phocae vitulinae is to the influence () of triglyceride
| Group | Dosage ml/kg | Number of animals n | x±SD.mg/g | The p value |
| Blank model contrast bifendate Adeps Phocae vitulinae | —— 0.2g 0.16g 2.2 4.3 8.6 | 8 8 8 8 8 8 | 2.37±0.76 5.92±0.68* 2.68±0.45 5.32±3.3 4.13±1.1 3.14±0.64 | —— P＜0.01 P＞0.05 p＜0.05 p＜0.01 |
Compare with the blank group
*P＜0.01
Adeps Phocae vitulinae is to the influence (two) of triglyceride
| Group | Dosage ml/kg | Number of animals n | x±SD.mg/g | The p value |
| Blank model contrast bifendate Adeps Phocae vitulinae | —— 0.2g 0.16g 2.2 4.3 8.6 | 8 8 8 8 8 8 | 2.25±0.24 3.65±0.59* 2.68±0.45 3.28±0.29 2.89±0.51 2.68±0.45 | —— P＜0.01 P＞0.05 p＜0.05 p＜0.01 |
Compare with the blank group
*P＜0.01
The result shows that the content of triglyceride of blank group is in range of normal value; The numerical value of model control group raises then very obvious; And positive controls and Adeps Phocae vitulinae 8.6ml/kg, the numerical value of 4.3ml/kg group all has obvious reduction, has significant differences and significant difference (p＜0.01, p＜0.05) with the analysis of model control group comparative statistics.
Conclusion
Adeps Phocae vitulinae is under 4.3ml/kg-8.6ml/kg dosage, continuously gastric infusion is 9 days, and the fatty liver to due to the rat DL-ethionine can significantly reduce the raising of content of triglyceride, can effectively prevent the generation of fatty liver, confirm that Adeps Phocae vitulinae has lipotropic preferably effect.
Cause the model of rat fat liver with the DL-ethionine, to Adeps Phocae vitulinae under 2.2ml/kg-8.6ml/kg dosage, 9 days pharmacodynamic action of continuous irrigation stomach is observed, and found that: this medicine can obviously suppress the DL-ethionine and cause increasing of rat triglyceride content.
Description of drawings
Below be description of drawings of the present invention,, can more be expressly understood the present invention by description of drawings and in conjunction with above-mentioned specific descriptions, specific as follows:
Accompanying drawing 1 is the uv absorption spectrogram of Adeps Phocae vitulinae of the present invention;
Accompanying drawing 2 is infrared absorption spectras of Adeps Phocae vitulinae of the present invention;
Accompanying drawing 3-5 is the gas chromatogram of the standard substance of the EPA methyl ester that uses among the present invention, DHA ethyl ester, DPA methyl ester;
Accompanying drawing 6-7 is the gas chromatogram of Adeps Phocae vitulinae of the present invention.
Claims (3)
1, a kind of Main Ingredients and Appearance is the application of the Adeps Phocae vitulinae of eicosapentaenoic acid, clupanodonic acid and docosahexenoic acid as the preparation medicine for treating fatty liver, in the wherein said Adeps Phocae vitulinae, the ratio of the content of eicosapentaenoic acid, docosahexenoic acid and clupanodonic acid is 1.00072%: 0.49247%: 1.09003% or 1.19101%: 0.58635%: 1.27373%.
2, Adeps Phocae vitulinae according to claim 1 is as the application of preparation medicine for treating fatty liver, and the dosage that it is characterized in that Adeps Phocae vitulinae in the described application is 4.3-10mL/Kg.
3, Adeps Phocae vitulinae according to claim 1 and 2 is as the application of preparation medicine for treating fatty liver, and the dosage that it is characterized in that Adeps Phocae vitulinae in the described application is 5-8.6mL/Kg.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011355174A CN1250230C (en) | 2001-09-30 | 2001-09-30 | Application of fur seal oil in preparation of medicine for treating fatty liver |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011355174A CN1250230C (en) | 2001-09-30 | 2001-09-30 | Application of fur seal oil in preparation of medicine for treating fatty liver |
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| Publication Number | Publication Date |
|---|---|
| CN1410073A CN1410073A (en) | 2003-04-16 |
| CN1250230C true CN1250230C (en) | 2006-04-12 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNB011355174A Expired - Fee Related CN1250230C (en) | 2001-09-30 | 2001-09-30 | Application of fur seal oil in preparation of medicine for treating fatty liver |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102755350B (en) * | 2012-08-01 | 2015-07-01 | 浙江金诺康生物制药有限公司 | Refined seal oil and application thereof to preparation of medicament for treating non-alcoholic fatty liver |
| GB201301626D0 (en) * | 2013-01-30 | 2013-03-13 | Dignity Sciences Ltd | Composition comprising 15-OHEPA and methods of using the same |
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