CN102335225B - Extract of rust-coloured crotalaria herb with root for treating cardiovascular disease, and preparation method and application thereof - Google Patents
Extract of rust-coloured crotalaria herb with root for treating cardiovascular disease, and preparation method and application thereof Download PDFInfo
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- CN102335225B CN102335225B CN2011103144824A CN201110314482A CN102335225B CN 102335225 B CN102335225 B CN 102335225B CN 2011103144824 A CN2011103144824 A CN 2011103144824A CN 201110314482 A CN201110314482 A CN 201110314482A CN 102335225 B CN102335225 B CN 102335225B
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- crotalariae ferrugineae
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Abstract
The invention provides an extract of rust-coloured crotalaria herb with root for treating cardiovascular disease, and a preparation method and application thereof. The extract of rust-coloured crotalaria herb with root comprises sterides and oleanolic acid which can effectively improve lipid levels of mice suffering from hyperlipemia, reduce serum total cholesterol, and low density lipoprotein cholesterin, and prevent and treat cardiovascular disease; and the preparation method for the extract is simple, slightly pollutes environment and is suitable for large-scale industrial production.
Description
Technical field
The present invention relates to a kind of Chinese medicine extract and relate in particular a kind of Herba Crotalariae Ferrugineae extract, preparation method and application that is used for the treatment of cardiovascular and cerebrovascular disease in the application of pharmaceutical field.
Technical background
Epidemiological study shows, dyslipidemia is the very important pathogenic risk factor of atherosclerosis.In population of China blood, there were the cholesterol of atherogenicity effect and triglyceride levels low than the westerner in the past, and have the High-density Lipoprotein-cholesterol of study of anti-atherogenic effect higher, be considered to suffer from atherosclerosis and the less one of the main reasons of coronary atherosclerotic heart disease (coronary heart disease).But in recent years the investigation of China's blood lipids in healthy people is found, men and women crowd's blood T-CHOL and triglyceride levels all rise than before, the corresponding increase of atherogenic danger.Therefore, find natural, safely and effectively regulate the blood fat living resources to the control cardiovascular and cerebrovascular disease significant.These disease cause of disease complexity, but with abnormalities of sugar/lipid metabolism, certain relation is all arranged.Hyperlipemia more and more receives people's concern as the main independent hazard factor that causes cardiovascular and cerebrovascular disease.Studies have shown that, correctly adjust the M & M that blood lipid level can reduce cardiovascular and cerebrovascular disease effectively.Most methods that reduce blood cholesterol levels need to strictly be kept on a diet and depend on medicine clinically, and the side effect of blood lipid-lowering medicine its application is subject to restriction to a certain degree.The correlational study of Herba Crotalariae Ferrugineae and extract thereof mainly concentrates on the anti-inflammation aspect at present, and Chinese patent (200710077676.0) discloses a kind of natural antibacterial composition extracted from Herba Crotalariae Ferrugineae and extracting method thereof; Wu Zhenjian etc. have reported about the research of Herba Crotalariae Ferrugineae flavone extract antibacterial activity in vitro.
Summary of the invention
Herba Crotalariae Ferrugineae bitter in the mouth, micro-acid, cold in nature, among the peoplely be used for the treatment of chronic cough expectorant blood, chronic nephritis, tonsillitis, lymphadenitis, treatment poison malignant boil, menoxenia etc.; By research, find, be rich in steroid research in Herba Crotalariae Ferrugineae and find that the sterol be rich in the Herba Crotalariae Ferrugineae of various dose can improve the blood lipid level of hyperlipemia in mice to some extent, reduce serum total cholesterol, low-density lipoprotein cholesterol and atherogenic index, rising serum High Density Lipoprotein Cholesterol level; In addition, its oleanolic acid be rich in has the cruel propylhomoserin level of the rats with liver cirrhosis brain homogenate of reduction, therefore be conducive to the control of hepatic encephalopathy.The present invention provides a kind of Herba Crotalariae Ferrugineae extract, preparation method and application particular content that is used for the treatment of cardiovascular and cerebrovascular disease to be on the basis of above-mentioned research:
Be used for the treatment of the preparation method of the Herba Crotalariae Ferrugineae extract of cardiovascular and cerebrovascular disease, comprise the following steps:
1) pulverize the Herba Crotalariae Ferrugineae crude drug, the dress post, use the 60-95% ethanol percolation, merges recovery extracting solution recovery ethanol system dry, obtains ethanol extract;
2) ethanol in the decompression recycling ethanol extracting solution to without the alcohol flavor, constantly adds water in the process that reclaims ethanol, and the material of separating out that makes to reclaim in ethanol process is suspension, obtains solution of extract;
3) with the extracting and washing solution of extract repeatedly of any one or two or more mixture in petroleum ether, ether, gasoline, n-butyl alcohol, n-amyl alcohol or benzene, till no suspended substance matter in mother solution and saponification foam, merge and reclaim organic facies, obtain the Herba Crotalariae Ferrugineae extract.
The condition of the described percolation of step 1) is: the percolation solvent adopts 75-90% ethanol, the 8-10 that percolation ration is medical material times volume.
The described merging recovery of step 3) organic facies obtains the Herba Crotalariae Ferrugineae extract and also passes through purification, and concrete steps are:
I. the heating organic facies, to lower than boiling point 1-2 ℃, has been recycled to material and has separated out, and obtains the crystallize mother solution;
II. the crystallize mother solution is placed in to water-bath or oil bath, with the per hour speed cooling of 3-5 ℃, to room temperature, filters and obtain crude extract;
III. use the absolute ethanol washing crude extract, to the precipitate surface, without oil substances, obtain the Herba Crotalariae Ferrugineae extract.
Prepare the Herba Crotalariae Ferrugineae extract that is used for the treatment of cardiovascular and cerebrovascular disease according to the described method of above-mentioned steps; Wherein, comprise steroidal composition and oleanolic acid in the Herba Crotalariae Ferrugineae extract.
The described application of Herba Crotalariae Ferrugineae extract in preparation treatment cardiovascular and cerebrovascular disease that is used for the treatment of cardiovascular and cerebrovascular disease.
Useful technique effect of the present invention is: the invention provides the blood lipid level that a kind of Herba Crotalariae Ferrugineae extract that is used for the treatment of cardiovascular and cerebrovascular disease can effectively improve hyperlipemia in mice, reduce serum total cholesterol, low-density lipoprotein cholesterol, prevention and Cardiovarscular; And method for preparing extractive technique provided by the present invention is simple, environmental pollution is little, is applicable to large-scale commercial production.
The specific embodiment
embodiment 1 is used for the treatment of the preparation of the Herba Crotalariae Ferrugineae extract of cardiovascular and cerebrovascular disease
Weighing Herba Crotalariae Ferrugineae crude drug 5.0kg, pulverize, and with the solvent 1L of 90% ethanol, infiltrates, and dress percolation post, add solvent to soak 12h, the beginning percolation, and the 8-10 that percolation ration is medical material times volume, take the basis for estimation that the sterol amount is the percolation volume in percolate, obtains percolation filtrate;
Selection for the percolation solvent can be selected 60-95%, and the solvent in this scope all can be realized the extraction to effective site in Herba Crotalariae Ferrugineae; But optimum solvent concentration is 75-90% ethanol;
Merge percolation filtrate, the ethanol in the decompression recycling ethanol extracting solution to without the alcohol flavor, constantly adds water in the process that reclaims ethanol, and the material of separating out that makes to reclaim in ethanol process is suspension, obtains solution of extract;
With the extracting and washing solution of extract repeatedly of any one or two or more mixture in petroleum ether, ether, gasoline, n-butyl alcohol, n-amyl alcohol or benzene, till no suspended substance matter in mother solution and saponification foam, merge and reclaim organic facies, obtain the Herba Crotalariae Ferrugineae extract, wherein the Herba Crotalariae Ferrugineae extract in should contain steroid material and oleanolic acid.
If lipid material content is higher in said extracted thing aqueous solution, the Herba Crotalariae Ferrugineae extract after causing extracting contains more lipid material and terpene substances, need to after extraction, add single step purification technique; Wherein the step of purifying process is:
I. the heating organic facies, to lower than boiling point 1-2 ℃, has been recycled to material and has separated out, and obtains the crystallize mother solution;
II. the crystallize mother solution is placed in to water-bath or oil bath, with the per hour speed cooling of 3-5 ℃, to room temperature, filters and obtain crude extract;
III. use the absolute ethanol washing crude extract, to the precipitate surface, without oil substances, obtain the Herba Crotalariae Ferrugineae extract.
The content of effective ingredient in should the Detection and Extraction position in each step of above-mentioned technique, wherein the detection method of each effective ingredient is:
The testing conditions of oleanolic acid is: chromatographic column is (4.6 mm * 150.0mm, 5 m); Mobile phase is methanol one water (V:V=90:10); Detecting wavelength is 220 nm; Sample size is 10 μ l; Flow velocity is 1.0 mL/min; Column temperature is 25 ℃.
The testing conditions of steroidal is: A, extract each operation stage extract, be dried to the thing solvent, residue adds the 1ml glacial acetic acid and dissolves, then adds 1ml vinegar and join, and drips one of concentrated sulphuric acid, displaing amaranth in the solution edge; B, extract each operation stage extract, be dried to the thing solvent, dissolve with the 1ml chloroform, add the 1ml concentrated sulphuric acid, chloroform layer is aobvious red, and the concentrated sulphuric acid layer has pistac.Above test shows that this plant is containing steroidal.
the experimentation of embodiment 2 Herba Crotalariae Ferrugineae extracts to hyperlipidemia model rat effect for reducing blood fat
The experimental animal male SD rat, 60, body weight 150～200g, clean level.
20 ℃～24 ℃ of SPF experimental animal room ambient temperatures, relative humidity 40%～70%, day alternates with night 12h/12h.
High lipid food 78.95% normal feedstuff, 1% cholesterol, 10% yolk powder, 10% Adeps Sus domestica, 0.05% bovine bile;
By the Ministry of Public Health function of health food, detect and the assessment technique standard: rat conforms after 3d, feed normal feedstuff 5d, overnight fasting is adopted tail blood, centrifugal (3000 rpm), get serum, survey serum TC, TG, HDL-C, according to the TC level, be divided at random 5 groups, be i.e. three dosage groups (0.33,0.66 and 2.00g/kg), a high lipid food matched group and a normal feedstuff matched group.Experimental period,, interior three dosage groups gave the tested material of high lipid food and various dose, and the high lipid food matched group gives high lipid food and distilled water, and the normal feedstuff matched group gives normal feedstuff and distilled water.30d in test weighs, and gets tail blood after fasting 16h, surveys serum TC, TG, HDL-C.
Statistical analysis adopts the SPSS10.0 statistical software to carry out variance analysis, and significance level is decided to be α=0.05.
The Herba Crotalariae Ferrugineae extract on Serum TC affect table 1
Annotate: each dosage group and hyperlipidemia model compare
* p<0.01,
* p<0.01
High lipid food control animals serum TC and normal feedstuff matched group comparing difference have statistical significance (
p<0.01), illustrate that this test hyperlipidemia model is successfully established.After test, the Herba Crotalariae Ferrugineae extract of 0.66,2.00 g/ kg dosage groups can make TC in rat blood serum reduce, with high lipid food matched group comparing difference have statistical significance (
p<0.01).
The affect table 2 of Herba Crotalariae Ferrugineae extract on rat blood serum TG
Annotate: each dosage group and hyperlipidemia model compare
* p<0.01,
* p<0.01
After test, the Herba Crotalariae Ferrugineae extract of 0.33,0.66,2.00 g/kg dosage groups can make TG in rat blood serum reduce, with high lipid food matched group comparing difference have statistical significance (
p<0
.05
, P<0.01), illustrate that the Herba Crotalariae Ferrugineae extract has reducing effect to the TG in rat blood serum.
The affect table 3 of Herba Crotalariae Ferrugineae extract on rat blood serum HDL-C
annotate: each group compares with the high lipid food matched group
p0.05
Serum HDL-C difference not statistically significant between each group, illustrate the Herba Crotalariae Ferrugineae extract to HDL-C without the rising effect.
This experimental result shows, 0.66, the Herba Crotalariae Ferrugineae extract of 2.00g/kg dosage group can make TC in rat blood serum obviously reduce, 0.33,0.66 and the Herba Crotalariae Ferrugineae extract of 2.00g/kg dosage group all can make TG in rat blood serum reduce, but the Herba Crotalariae Ferrugineae extract has no significant effect HDL-C, the reduction blood fat of Herba Crotalariae Ferrugineae extract is described; Adjust the M & M that blood lipid level can reduce cardiovascular and cerebrovascular disease effectively.
Claims (4)
1. a preparation method that is used for the treatment of the Herba Crotalariae Ferrugineae extract of cardiovascular and cerebrovascular disease is characterized in that: comprise the following steps:
1) pulverize the Herba Crotalariae Ferrugineae crude drug, the dress post, use the 60-95% ethanol percolation, merges and reclaim extracting solution recovery ethanol to dry, obtains ethanol extract;
2) ethanol in the decompression recycling ethanol extracting solution to without the alcohol flavor, constantly adds water in the process that reclaims ethanol, and the material of separating out that makes to reclaim in ethanol process is suspension, obtains solution of extract;
3) with the extracting and washing solution of extract repeatedly of any one or two or more mixture in petroleum ether, ether, gasoline, n-butyl alcohol, n-amyl alcohol or benzene, till no suspended substance matter in mother solution and saponification foam, merge and reclaim organic facies, obtain the Herba Crotalariae Ferrugineae extract;
Described merging recovery organic facies obtains the Herba Crotalariae Ferrugineae extract and also passes through purification, and concrete steps are:
I. the heating organic facies, to lower than boiling point 1-2 ℃, has been recycled to material and has separated out, and obtains the crystallize mother solution;
II. the crystallize mother solution is placed in to water-bath or oil bath, with the per hour speed cooling of 3-5 ℃, to room temperature, filters and obtain crude extract;
III. use the absolute ethanol washing crude extract, to the precipitate surface, without oil substances, obtain the Herba Crotalariae Ferrugineae extract.
2. the preparation method that is used for the treatment of the Herba Crotalariae Ferrugineae extract of cardiovascular and cerebrovascular disease according to claim 1, it is characterized in that: the condition of the described percolation of step 1) is: the percolation solvent adopts 75-90% ethanol, the 8-10 that percolation ration is medical material times volume.
3. a Herba Crotalariae Ferrugineae extract that is used for the treatment of cardiovascular and cerebrovascular disease, it is characterized in that: method according to claim 1 and 2 prepares the Herba Crotalariae Ferrugineae extract that is used for the treatment of cardiovascular and cerebrovascular disease; Wherein, comprise steroidal composition and oleanolic acid in the Herba Crotalariae Ferrugineae extract.
4. the application of Herba Crotalariae Ferrugineae extract in preparation treatment cardiovascular and cerebrovascular diseases medicament that is used for the treatment of cardiovascular and cerebrovascular disease according to claim 3.
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| Title |
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| 响铃草、松针、甘松有效成分研究;张旭;《成都中医药大学博士研究生论文》;20061231;11，29 * |
| 张旭.响铃草、松针、甘松有效成分研究.《成都中医药大学博士研究生论文》.2006,11，29. |
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