CN101618096A - Madder extract and preparation method and application thereof - Google Patents
Madder extract and preparation method and application thereof Download PDFInfo
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- CN101618096A CN101618096A CN200910065554A CN200910065554A CN101618096A CN 101618096 A CN101618096 A CN 101618096A CN 200910065554 A CN200910065554 A CN 200910065554A CN 200910065554 A CN200910065554 A CN 200910065554A CN 101618096 A CN101618096 A CN 101618096A
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- radix rubiae
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Abstract
本发明涉及一种茜草提取物及其制备方法和应用，其中茜草提取物由以下方法制备获得：将茜草根茎经粉碎，用乙醇在室温下渗漉提取；回收乙醇，得到浸膏；将浸膏溶于水中，加热使其完全溶解；然后用乙酸乙酯萃取，浓缩得到茜草乙酸乙酯提取物。实验表明：茜草乙酸乙酯提取物及化合物茜素能有效降低四氧嘧啶致高血糖小鼠的血糖值、GSP值、TG值和组织匀浆中的MDA含量，且能显著的升高小鼠肝细胞的SOD活力值以及增加组织肝糖原含量。证明茜草乙酸乙酯提取物和茜素具有很好的降血糖作用。The present invention relates to a madder extract and its preparation method and application, wherein the madder extract is prepared by the following method: crushing the rhizome of madder, extracting by percolating with ethanol at room temperature; recovering the ethanol to obtain an extract; extracting the extract Dissolved in water, heated to dissolve completely; then extracted with ethyl acetate, concentrated to obtain Rubia ethyl acetate extract. Experiments show that the extract of Rubia ethyl acetate and the compound alizarin can effectively reduce the blood glucose level, GSP value, TG value and MDA content in tissue homogenate of mice with hyperglycemia induced by alloxan, and can significantly increase the SOD activity value of hepatocytes and increase tissue liver glycogen content. It is proved that Rubia ethyl acetate extract and alizarin have good hypoglycemic effect.
Description
Technical field
The present invention relates to a kind of preparation method of Radix Rubiae ethyl acetate extract and prepared the application of blood sugar lowering with the treatment diabetes medicament; Relate in particular to the application of a kind of this position separating compound alizarin aspect preparation blood sugar lowering and treatment diabetes medicament.
Background technology
Diabetes be a kind of be feature with the hyperglycemia, metabolism disorder for the performance, with the closely-related systemic disease of insulin secretion, be the clinical syndrome that causes owing to the h and E factor interaction.Absolute or relative deficiency and target tissue reduce insulin sensitivity because of insulin secretion, cause a series of metabolism disorders such as sugar, albumen, fat, power and water Xie Zhi.Diabetes mainly are divided into two types: type i diabetes (being insulin dependent diabetes mellitus (IDDM)) and type ii diabetes (being non-insulin-dependent diabetes mellitus), the latter accounts for the 99%-95% of patient's sum.The clinical manifestation of diabetes is more than three (polyphagia, polydipsia, polyurias), few (weight loss), a hyperglycemia symptom.The complication that diabetes cause (as acute complicationses such as cardiovascular, kidney, retina and neural chronic complicating diseases, diabetic ketoacidosis, hyperosmolar nonketotic diabetic comas) is the main cause that causes death.All diabetic complications can be by strict glycemic control, from preventing to the full extent, if diabetics can be controlled at the blood sugar level of oneself near in the normal range of blood sugar, then diabetic complication just can not occur, and perhaps occurs than later and degree is also lighter.Diabetes are one of modal epidemic diseases of world wide, have become the non-infective disease of the third-largest serious threat human health after cardiovascular, tumor.According to The World Health Organization (WHO) report, nineteen ninety-five the diabetics made a definite diagnosis of the whole world about 1.35 hundred million, and IDF 2006 annual reports whole world diabetics has reached 2.46 hundred million and predicts diabetics in 2025 and will reach 3.88 hundred million in the recent period.China had become diabetes second big country in 2003, had 2,380 ten thousand diabeticss, and the expert estimates, will reach 6,000 ten thousand to China's number of the infected in 2010.Diabetes become the 3rd " the healthy killer " who threatens the mankind, so prevention and treatment diabetes have become global health care problem.
The treatment of diabetes medicine mainly concentrates on sulphanylureas and biguanides for a long time based on Western medicine for many years.But these medicines exist untoward reaction such as hypoglycemia and gastrointestinal reaction, thereby the antidiabetic drug of searching high-efficiency low-toxicity is still the target that medical expert pursues.Because that Chinese medicine and natural plant have is comparatively safe and cheap, action temperature and lastingly and can delay advantage such as complication, can yet be regarded as and seek an important channel of blood sugar lowering new drug, therefore develop Chinese medicine and natural plant and can provide Remedies for diabetes source widely as antidiabetic drug.
Radix Rubiae is a rubia, popular name Herba Chenopodii Hybridi, Radix Rubiae Yunnanensis, HUOXUECAO, saw grass, draws rattan, Radix seu Caulis Schisandrae Sinensis, is the dry root and rhizome of Maguireothamnus speciosus Radix Rubiae (Rubia cordifolia L.).Kind surplus the Rubiaceae Rubia plant whole world has 60 mainly is distributed in Asia, Europe, America and Africa.There is 24 kind of 10 mutation in China, and the various places that spread all over the country are especially with the southwest most species.Radix Rubiae is widely distributed, but be the main producing region still so far with Shaanxi, Shanxi and Henan, Radix Rubiae bitter in the mouth cold in nature, GUIXIN Liver Channel, have cooling blood for hemostasis, expelling wind and removing dampness, dispel become silted up stimulate the menstrual flow, the effect of blood circulation promoting and blood stasis dispelling, eliminating phlegm and stopping cough, cure mainly haematemesis, epistaxis, metrorrhagia due to blood-heat, amenorrhea, traumatic injury.Mainly contain water miscible ring peptide series matter, liposoluble constituent anthraquinone and glucosides, naphthoquinone and glucosides thereof in the Radix Rubiae, contain terpenoid, cupreol, fatty acid cpds and trace element in addition.
Alizarin is the anthraquinone analog compound that extracts from Radix Rubiae (Rubia Cordifolia L.) root, separates, formal name used at school 1,2-dihydroxyanthraquinone.Pharmacological research shows that alizarin has significant bacteriostasis, and this chemical compound others pharmacologically active is not seen the research report.
Summary of the invention
The present invention also aims to provide a kind of Madder extract.
The present invention also aims to provide a kind of preparation method of Radix Rubiae ethyl acetate extract.
Simultaneously, the present invention also aims to provide the application of a kind of Radix Rubiae ethyl acetate extract aspect preparation treatment diabetes medicament.
The present invention also aims to provide a kind of from the Chinese medicine Radix Rubiae isolated compound alizarin, this chemical compound can be used for preparing hypoglycemic drug.
To achieve these goals, technical scheme of the present invention has adopted a kind of Madder extract, is prepared by following method: with the Radix Rubiae rhizome through pulverizing, with ethanol at room temperature percolation extract; Reclaim ethanol, obtain extractum; Extractum is soluble in water, and heating is dissolved it fully; Use ethyl acetate extraction then, concentrate and obtain the Radix Rubiae ethyl acetate extract.
Described Radix Rubiae rhizome drying is handled.
Described ethanol volume fraction is 95%.
Simultaneously, technical program of the present invention also lies in adopting a kind of preparation method of Radix Rubiae ethyl acetate extract, may further comprise the steps: the Radix Rubiae stem is dry, pulverize, with volume fraction be 95% ethanol at room temperature percolation extract, recovery ethanol obtains extractum; Extractum is soluble in water, and heating is dissolved it fully; Use ethyl acetate extraction then, concentrate and obtain ethyl acetate extract.
Technical scheme of the present invention has also adopted a kind ofly separates the chemical compound alizarin German side method that obtains from ethyl acetate extract, concrete steps are: ethyl acetate extract is through 200～300 order silica gel column chromatographies, petroleum ether-acetone gradient elution (95: 5～9: 1), obtain 4 parts, wherein the 3rd part is through the silica gel H column chromatography, petroleum ether-acetone (15: 1) eluting, detect merging with TLC, through Sephadex LH-20 column chromatography, the acetone eluting obtains the chemical compound alizarin.
Further, technical scheme of the present invention has adopted a kind of Radix Rubiae ethyl acetate extract at the preparation blood sugar lowering and be used for the treatment of application aspect the diabetes medicament.
Simultaneously, also adopted a kind of chemical compound alizarin at the preparation blood sugar lowering and be used for the treatment of application aspect the diabetes medicament.
Described chemical compound alizarin is from any crude drug that contains alizarin, preferably from the ethyl acetate extract in the Chinese medicine Radix Rubiae crude drug.
Radix Rubiae distributes very extensive at occurring in nature, this just makes raw material of the present invention be easy to get, and cost is very low.Experiment shows: Radix Rubiae ethyl acetate extract and sharp its have good hypoglycemic activity through the alizarin that further extraction separation obtains.
With alloxan inducing mouse β cell injury, after causing diabetes model, irritate stomach with Radix Rubiae ethyl acetate extract and alizarin, detect SOD and MDA value in blood glucose value, GSP value, TG value, hepatic glycogen content and the liver tissue homogenate in the mice serum, experiment shows: Radix Rubiae ethyl acetate extract and chemical compound alizarin can effectively reduce the MDA content in blood glucose value, GSP value, TG value and the tissue homogenate that alloxan causes the hyperglycemia mice, and hepatic glycogen content is organized in the SOD energy value and the increase of the mouse liver cell that can raise significantly.Proof Radix Rubiae ethyl acetate extract and alizarin have good hypoglycemic activity.
By adopt Radix Rubiae ethyl acetate extract that method of the present invention handles gained with and separated product chemical compound alizarin, can make various preparations, comprise: suppository, tablet, pill, granule, membrane, microcapsule, drop pill, aerosol, medicated wine, syrup, oral liquid etc.
The specific embodiment
Embodiment 1
Present embodiment is a Madder extract---the extracting method of ethyl acetate extract specifically may further comprise the steps: get the Radix Rubiae dry product, pulverize, extract with percolation under the 95% ethanol room temperature, reclaim ethanol, obtain extractum; Extractum is soluble in water, and heating is dissolved it fully; Use ethyl acetate extraction then, concentrate and obtain ethyl acetate extract.
Embodiment 2
Present embodiment is the separated product of Radix Rubiae ethyl acetate extract---the separation method of chemical compound alizarin, body may further comprise the steps: ethyl acetate extract is through 200～300 order silica gel column chromatographies, petroleum ether-acetone gradient elution (95: 5～9: 1), obtain 4 parts, wherein the 3rd part is through the silica gel H column chromatography, petroleum ether-acetone (15: 1) eluting, detect merging with TLC, through Sephadex LH-20 column chromatography, the acetone eluting obtains the chemical compound alizarin.
Embodiment 3
Present embodiment is the treatment diabetes effect experiment of Radix Rubiae ethyl acetate extract and chemical compound alizarin.
Method: mice alloxan diabetes model
Principle: alloxan (Alloxan) is the beta Cell of islet toxic agent, optionally damages the beta Cell of islet of multiple animal, causes diabetes.Because its round modeling method is easy, the formation rate height is the most frequently used diabetes animal model.In the stability study for alloxan mice diabetes model, mainly be by alloxan solution, after injection due to diabetes, detect blood glucose value in the mice serum in the certain hour.During diabetes, reflection past 1-2 week average blood sugar level and medicine can raise to GSP (saccharifying serum albumin) content of the influence of nonenzymatic glycosylation effect in the body, and simultaneously, some enzymatic activity in the body in the glycogen building-up process is low, glycogen is synthetic to be reduced, and shows as hepatic glycogen content and reduces.In addition, diabetes can cause simultaneously the oxidative damage of tissue, show as that superoxide dismutase (SOD) in the hepatic tissue reduces and the rising of the content of malonaldehyde (MDA).Also can cause the lipid metabolism obstacle, show as the rising of serum triglycerides (TG).
Instrument: UV-2000 type ultraviolet-uisible spectrophotometer (but You Ni Shanghai Instr Ltd.), CS-H1 type blender (Beijing is won and encouraged positive scientific ﹠ technical corporation), the biochemical incubator (Shanghai one permanent Science and Technology Ltd.) of LRH-150, Multiskan MK3 microplate reader (thermoelectric Shanghai Instr Ltd.), TGL-16 high speed centrifuge (big instrument plant in Jintan City, Jiangsu), the tissue refiner of 985370-395 type (BIOSREC, Mexico), all size pipettor etc.
Reagent: Alloxan (alfa asear company; lot number: 2244-11-3) acarbose tablet (Zhongmei Huadong Pharmaceutical Co., Ltd. Hangzhou; lot number: 080105); Coomassie brilliant blue G250 (packing factory of Shanghai chemical reagents corporation; lot number: 20050115); (Shanghai Rongsheng Bioisystech Co., Ltd, lot number: 20080705) GSP, TG, hepatic glycogen detection kit (be Nanjing and build up bio-engineering research institute), other reagent are analytical pure to the glucose assays test kit.
Experimental technique:
Mice (male, body weight: behind 22 ± 2g) the adaptation 7d, divide in groups at random, 10 of blank groups, another group is diabetes modeling group.Diabetes modeling group causes diabetes by disposable tail vein injection 4% alloxan of 80mg/kg after fasting be can't help water 12-16h hour.
Water 6h is can't help in fasting after 96h hour, weighs, the eye socket vein gets blood, separation of serum.With blood glucose value greater than the standard of 11.1mmol/L as successful modeling, according to of the grouping of blood glucose value homeostatic principle with the modeling success, 11 every group.
Administration group in grouping back begins to irritate the corresponding medicine of stomach every day, the acarbose positive controls is irritated stomach 75mg/kg every day, Radix Rubiae ethyl acetate extract group is irritated stomach 750mg/kg every day, the alizarin group is irritated stomach 50mg/kg every day, normal control group and model group are irritated stomach corresponding C MC-Na every day, fasting 6h behind the administration 8d, pluck eyeball and get blood system from the determination of serum fasting blood sugar, GSP content, TG content, isolating liver weighs, get a part (about 80mg) and measure hepatic glycogen content, and the liver of the about 300mg of weighing and about 30mg measures MDA and SOD with the liver homogenate liquid of 0-3 ℃ of normal saline homogenate (surveying MDA) and 1% (the surveying SOD) that become 10%, and Coomassie brilliant blue is except that albumen.Other gets spleen, kidney is weighed and calculated each organ index.
The mensuration of blood glucose, GSP, TG, hepatic glycogen content, SOD and MDA is all carried out according to the requirement of test kit, and experimental data x ± s represents that experiment statistics adopts the spss11.0 statistical software to carry out one factor analysis of variance.
The result:
(1) Radix Rubiae ethyl acetate extract and chemical compound alizarin the results are shown in Table 1 to the influence of alloxan diabetes mouse blood sugar value, GSP content, hepatic glycogen content, show in the table, the blood glucose value of Radix Rubiae ethyl acetate extract group and alizarin group all is lower than model group and the preceding value of administration, demonstrate significant hypoglycemic activity, and GSP content is starkly lower than model group, and hepatic glycogen content is apparently higher than model group, and data difference has the significance meaning.
(2) Radix Rubiae ethyl acetate extract and alizarin the results are shown in Table 2 to the influence of the oxidability of alloxan diabetes mice, data show, the SOD level of model group is starkly lower than the blank group, and the SOD level of Radix Rubiae ethyl acetate extract and alizarin is apparently higher than model group, alizarin even be higher than the blank group; And the MDA level is starkly lower than model group.Show that Radix Rubiae ethyl acetate extract and extract alizarin have certain antioxidation, can treat the relevant oxidative damage of diabetes.
(3) Radix Rubiae ethyl acetate extract and alizarin influence table 2 data show to the blood lipid level of alloxan diabetes mice, the TG content of model group is apparently higher than the blank group, and the TG content of Radix Rubiae ethyl acetate extract and alizarin is starkly lower than model group, and be lower than the blank group, demonstrate the effect for reducing blood fat that is better than positive control drug, show that the dysbolism of blood fat that they cause for diabetes has certain therapeutical effect.
Table 1 alizarin ethyl acetate extract and alizarin are to the therapeutical effect of diabetes model (x ± s)
Each group compares with the blank group:
△P＜0.05,
△ △P＜0.01,
△ △ △P＜0.001;
Each group compares with model group:
*P＜0.05,
*P＜0.01,
* *P＜0.001;
Each group compares with positive controls:
#P＜0.05,
##P＜0.01,
###P＜0.001
Table 2 alizarin ethyl acetate extract and alizarin are to diabetes model
Antioxidation and effect for reducing blood fat (x ± s)
Each group compares with the blank group:
△P＜0.05,
△ △P＜0.01,
△ △ △P＜0.001;
Each group compares with model group:
*P＜0.05,
*P＜0.01,
* *P＜0.001;
Experimental result shows, diabetic model group compares with blank group matched group, blood glucose value is kept higher level in the serum, the GSP value all obviously raises, hepatic glycogen content reduces, and illustrates that institute's diabetes model of making is stable, and the MDA value all obviously raises in TG value and the hepatocyte, the SOD value reduces, and illustrates that diabetes have caused complication such as relevant oxidative damage and disorders of lipid metabolism.
Radix Rubiae ethyl acetate extract and alizarin group all have the blood glucose value that reduces diabetic mice, GSP content,, hepatic glycogen content in the rising hepatic tissue, as seen Radix Rubiae ethyl acetate extract and alizarin all have good hypoglycemic activity, can develop to be used for treatment of diabetes; Simultaneously, Radix Rubiae ethyl acetate extract and alizarin obviously reduce MDA value in diabetic mice serum TG content and the tissue, promote the SOD activity level of mice simultaneously.As seen alizarin ethyl acetate extract and chemical compound alizarin also have the effect of auxiliary treating diabetes such as fine antioxidation and blood fat reducing.
Embodiment 4
Present embodiment is the preparation method of Radix Rubiae ethyl acetate extract tablet:
Preparation method: get above-mentioned Radix Rubiae ethyl acetate extract 300g, the ethanol of adding an amount of 80% is made the wetting agent mixing and is made soft material behind adding starch 100g, the arabic gum 15g, under 60 ℃ of temperature, be dried to water content below 3%, then the thing of oven dry was pulverized 14 mesh sieves, after adding magnesium stearate 5g and Pulvis Talci 20g mixing at last, cross 12 mesh sieve granulate, tabletting, quality inspection, coating are made 1000.According to being grown up each 5 three times for each person every day.
Embodiment 5
Present embodiment is the preparation method of chemical compound alizarin tablet, and is specific as follows:
Preparation method: get above-claimed cpd alizarin 162g, with 60g starch mixing, the starch slurry 24g with 10% makes soft material, after the granulation of 14 mesh sieves, put 70 ℃～80 ℃ dry backs in 12 order granulate, behind adding dried starch 23g and the magnesium stearate 3g mixing, tabletting, quality inspection, coating are made 1000.According to being grown up each 2 three times for each person every day.
Embodiment 6
Present embodiment is the preparation method of Radix Rubiae ethyl acetate extract capsule, and is specific as follows:
Preparation method: get Radix Rubiae ethyl acetate extract 450g, divide on the drip pan of shop, put into 40-50 ℃ of baking oven and be dried to water content below 3%, block with oven dry is broken into powder below 8 orders then, adds 10% starch slurry and makes soft material in right amount, after the drying, granulate with 8 mesh sieves, directly incapsulate according to 0.4g powder/grain, make 1000 capsules, packing.Radix Rubiae ethyl acetate extract capsule is taken three times for each person every day, each 3 capsules.
Embodiment 7
Present embodiment is the preparation method of the capsule of chemical compound alizarin, and concrete steps are as follows:
Get chemical compound alizarin 90g,, add 10% starch slurry and make soft material in right amount, after granulating with 14 mesh sieves, put 70 ℃～80 ℃ dry backs and granulate, directly incapsulate, make 1000 capsules, pack according to 0.4g powder/grain with 8 mesh sieves with 40g starch mixing.Chemical compound alizarin capsule is taken three times for each person every day, each 1 capsules.
It should be noted last that: above embodiment is only in order to explanation, and unrestricted technical scheme of the present invention, although the present invention is had been described in detail with reference to the foregoing description, those of ordinary skill in the art is to be understood that: still can make amendment or be equal to replacement the present invention, and not breaking away from any modification or partial replacement of the spirit and scope of the present invention, it all should be encompassed in the middle of the claim scope of the present invention.
Claims (9)
1, a kind of Madder extract is characterized in that: prepared by following method: with the Radix Rubiae rhizome through pulverizing, with ethanol at room temperature percolation extract; Reclaim ethanol, obtain extractum; Extractum is soluble in water, and heating is dissolved it fully; Use ethyl acetate extraction then, concentrate and obtain the Radix Rubiae ethyl acetate extract.
2, Madder extract according to claim 1 is characterized in that: described Radix Rubiae rhizome drying is handled.
3, Madder extract according to claim 1 is characterized in that: described ethanol volume fraction is 95%.
4, a kind of preparation method of Madder extract is characterized in that: may further comprise the steps: the Radix Rubiae stem is dry, pulverize, with volume fraction be 95% ethanol at room temperature percolation extract, recovery ethanol obtains extractum; Extractum is soluble in water, and heating is dissolved it fully; Use ethyl acetate extraction then, concentrate and obtain ethyl acetate extract.
5, the preparation method of Madder extract according to claim 4 is characterized in that: the ethyl acetate extract that makes is separated obtaining the chemical compound alizarin.
6, the preparation method of Madder extract according to claim 5, it is characterized in that: may further comprise the steps: ethyl acetate extract is through 200～300 order silica gel column chromatographies, petroleum ether-acetone gradient elution (95: 5～9: 1), then through the silica gel H column chromatography, petroleum ether-acetone (15: 1) eluting detects merging with TLC, through Sephadex LH-20 column chromatography, the acetone eluting obtains the chemical compound alizarin.
7, the preparation method of Madder extract according to claim 6 is characterized in that: obtain 4 parts behind gradient elution, wherein the 3rd position is through the silica gel H column chromatography.
8, the Radix Rubiae ethyl acetate extract is at the preparation blood sugar lowering and be used for the treatment of application aspect the diabetes medicament.
9, the chemical compound alizarin is at the preparation blood sugar lowering and be used for the treatment of application aspect the diabetes medicament.
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