CN101810793B - Application of corn stigma flavone in preparing medicament for curing hyperlipidemia - Google Patents
Application of corn stigma flavone in preparing medicament for curing hyperlipidemia Download PDFInfo
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Abstract
The invention relates to application of corn stigma flavone in preparing a medicament for curing hyperlipidemia and losing weight, belongs to the field of medicaments, and particularly relates to new application of the corn stigma flavone in preparing the medicament for curing the hyperlipidemia and losing weight. The application has the advantages that: the corn stigma flavone is a pure natural medicament and can reduce blood fat and lose weight, so the blood fat reducing mechanism is not a single link but a result of multi-aspect and multi-target-spot combined action. The corn stigma flavone, used as the medicament for preventing and curing the hyperlipidemia or complicating diseases thereof and losing weight, has important practical significance and can add the new application of plantresources to drive regional economic development.
Description
Technical field
The invention belongs to field of medicaments, especially refer to the new pharmaceutical uses of Stigma Maydis flavone.
Background technology
Contain the pharmacological action of the Stigma Maydis of flavone component
1 antioxidation
1.1 extract antioxidant activity
Liu's equality extracts Stigma Maydis ethanol extraction reuse chloroform, ethyl acetate, n-butyl alcohol and methanol respectively, obtains 5 different components of polarity, and relatively these 5 kinds of different solvents extracts are in diphenyl picryl phenylhydrazine (DPPH) system and at Fe
2+Bring out the antioxidant activity in lipovitellinin polyunsaturated fatty acid (PUFA) the peroxidating system.The result shows: in the DPPH system, the antioxidant activity of n-butyl alcohol extract is the strongest, and chloroform extract takes second place; At Fe
2+Bring out in the lipoprotein PUFA peroxidating system, the antioxidant activity of chloroform extract is the strongest, and acetic acid ethyl ester extract takes second place.Can find out that thus same group of extract is in different test systems, the antioxidant activity order is different, so must consider concrete test system to the evaluation of antioxidant activity.Remain further research for the mechanism of Stigma Maydis extract antioxidation and the factor that influences antioxidant activity.
Z.A.Maksimoviand reports ripe Stigma Maydis methanolic extract to lipid peroxidation system Wheat Protein, and the strongest active position is medium oils concentrate, comprises phenolic acids, flavonoid and analog thereof.
Employing DPPH methods such as Fang Min and FRAP analytic process have been assessed the antioxidant activity of Stigma Maydis ethanol extraction.The result shows that the different concentration ethanol extract demonstrates different antioxidant activity, dries the Stigma Maydis to constant weight with the lixiviate of five kinds of different concentration ethanol solution, measures its antioxidant activity with two kinds of methods, and the oxidation resistance of 80% ethanol extraction is the strongest.The antioxidation of Stigma Maydis total flavones is obvious, and the clearance rate of DPPH free radical is reached 77.18%, and TAC FRAP value is 691.04 μ mol/L.Make an experiment with fresh corn silk, find that the dehydrated alcohol extraction thing removing DPPH free radical ability and the TAC of fresh corn silk is all the strongest.
WU YANAN utilizes hexichol for bitterness acyl group free radical system, H
2O
2/ Fe
2+The hydroxyl radical free radical system, the ultra-oxygen anion free radical system that produce are studied the ability of the external removing free radical of Stigma Maydis flavone gruff bring up substance, ethyl acetate and n-butyl alcohol extract, and are contrast with the measured value of rutin.Result of the test is: the ability of removing the DPPH free radical is followed successively by: Stigma Maydis flavone gruff bring up substance＞n-butyl alcohol extract＞acetic acid ethyl ester extract.Concentration and clearance rate that each sample reaches maximal clearance are respectively: 1.2,2g/L and 93%, 78% and 71%; The ability of removing hydroxy radical is followed successively by: Stigma Maydis flavone gruff bring up substance＞acetic acid ethyl ester extract＞n-butyl alcohol extract, and when sample solution concentration was 20g/L, it is maximum that clearance rate reaches, and is followed successively by 92.04%, 82.6% and 56.5%; The ability of removing ultra-oxygen anion free radical is followed successively by: Stigma Maydis flavone gruff bring up substance＞acetic acid ethyl ester extract＞n-butyl alcohol extract, when the concentration of sample was 20g/L, it is maximum that clearance rate reaches, and is followed successively by 94.8%, 90% and 58%.The result shows that the effect of each sample removing hydroxy radical and ultra-oxygen anion free radical is superior to the effect of rutin, and the Stigma Maydis flavone has the ability of removing free radical preferably.
Xu Gang has studied from the different parts of corn and has extracted Flavonoid substances, and its antioxygenic property has been carried out comparative study.The result shows: Flavonoid substances content is maximum in the Stigma Maydis, has reached 2.106%; Next is leaf of Semen Maydis 1.013%, corn cob 0.662%, and niblet content is minimum to be 0.411%.Through comparing with TBHQ, VC tea polyphenols, find that other threes show good antioxygenic property except that niblet, its antioxygenic property strengthens with the increase of extract concentrations.
Niu Peng flies to wait the flavone compound that passes through to compare opposed polarity in the Stigma Maydis to the hydroxyl radical free radical generation structure and to Fe
2+Bring out the inhibitory action of lipoprotein polyunsaturated fatty acid (PUFA) peroxidating system, studied the antioxidant activity (AOA) of opposed polarity flavone compound.The result shows that the antioxidant activity of the flavone compound of opposed polarity has than big difference.In the hydroxyl radical free radical generation structure, antioxidant activity is followed successively by chloroform＞n-butyl alcohol＞ethyl acetate in proper order.And at Fe
2+Bring out in the lipoprotein PUFA peroxidating system, antioxidant activity is followed successively by chloroform＞ethyl acetate＞n-butyl alcohol in proper order, in strict accordance with the ascending sequence arrangement of polarity.Find that simultaneously even the identical flavone compound of polarity, in different antioxidant system, its antioxidant activity also has tangible difference.
The filial piety of relaxing adds 0.01% extract (Stigma Maydis total flavones), ascorbic acid, citric acid, hexamethylene diamine base tetraacethyl disodium (EDTA) respectively with report in Adeps Sus domestica.Every its POV value of measuring at a distance from several days, non-oxidizability is strong and weak in proper order as follows in four kinds of reagent as a result: EDTA＞extracting solution＞citric acid＞bad hematic acid.The antioxidant activity that draws extract is more intense.
1.2 effective ingredient antioxidant activity
Ren Shuncheng etc. [utilize thiobarbituricacid method (TBA method), conjugated diene detection method (ultraviolet method) that the snperoxiaized ability of lipotropism plastid of from Stigma Maydis, separating 2 flavonoid ax-5 obtaining " methane-3 '-methoxymaysin and ax-4 "-OH-3 '-methoxymaysin is detected. the result shows that the former has stronger inhibition ability than the latter to the generation of conjugated diene (CD-POV), all shows certain dose-effect relationship.Stigma Maydis flavonoid monomer is to the inhibition ability of malonaldehyde (MDA) with to the inhibition ability basically identical of CD-POV, and the time that needs when reaching maximum than CD-POV when just MDA reaches maximum is longer, shows antioxidant activity.
What Stigma Maydis flavonoid chemical constituent was studied both at home and abroad in recent years is many, and the antioxidation aspect has obtained certain achievement.Research has been found same group of extract in different test systems, and the antioxidant activity that is showed is different, so must consider concrete test system to the evaluation of its antioxidant activity.Select for use which kind of evaluation method the most the problem such as mechanism and influence factor of science and Stigma Maydis extract antioxidation all await further further investigation.
2 Stigma Maydis water extract are to the effect of blood viscosity
Stigma Maydis total flavones such as Gao Nina can reduce blood stasis animal pattern whole blood viscosity and blood plasma viscosity, improve blood circulation.Stigma Maydis total flavones shows the blood plasma viscosity observation that influences of syndrome of blood stasis animal pattern blood plasma viscosity; Model group animal blood slurry viscosity (1.6774 ± 0.0826) is than the obvious rising of blank group (1.2108 ± 0.1069) (P＜0.01); Compare with model group; Stigma Maydis total flavones high dose group animal blood slurry viscosity (1.3642 ± 0.1096) obviously reduces (P＜0.05); And dose groups (1.4552 ± 0.1138) and low dose group (1.5220 ± 0.1871) and model group zero difference (P＞0.05) in the positive drug control group (1.4335 ± 0.1127), Stigma Maydis total flavones show that Stigma Maydis total flavones can reduce blood stasis animal pattern blood plasma viscosity.
3 Stigma Maydis water extract have effect for reducing blood fat
Miura T report Stigma Maydis water extract can obviously reduce normal mouse and experiment mice blood middle cholesterol content.Can significantly reduce triglyceride in the normal mouse blood.Content of triglyceride in the experiment mice blood is not influenced.Results suggest, the effect of Stigma Maydis water extract cholesterol reducing are owing to stop cholesterol synthetic at liver.
4 Stigma Maydis preparations are transferred blood fat
The fluffy oral liquid of beautiful a kind of reed mentioned in ancient books of developments such as Xu Wei is a compound Chinese medicinal preparation, is made up of Herba Salsolae Collinae, Stigma Maydis, charred Cortex Eucommiae, Radix Scutellariae 4 flavor Chinese medicines, has to bring high blood pressure down the effect of blood lipid regulation, swelling diuretic.This experiment shows that the fluffy oral liquid of beautiful a kind of reed mentioned in ancient books feeds for a long time with serum TC that high lipid food was caused, TG, the rising of LDL-C, the reduction of HDL-C rat obvious inhibitory action is arranged; To forming the rat of hyperlipemia; TC, TG, LDL-C level are obviously reduced; And make the HDL-C level that rising to a certain degree arranged, show that the fluffy oral liquid of beautiful a kind of reed mentioned in ancient books has tangible prevention and therapeutical effect to experimental hyperlipemia in rats.In addition, acute toxicity test and chronic toxicity test show that all the fluffy oral liquid toxicity of beautiful a kind of reed mentioned in ancient books is lower.Therefore, the fluffy oral liquid of beautiful a kind of reed mentioned in ancient books is having application promise in clinical practice aspect accent blood fat and the control AS.
The progress of Chinese medicine blood fat reducing
The research of 1 Chinese traditional medicines depressing lipid
Have the Chinese herbal medicine of effect for reducing fat have 90 surplus kind, like Fructus Crataegi, Rhizoma Alismatis, Radix Salviae Miltiorrhizae pellet, Radix Et Rhizoma Rhei, Semen Cassiae, Radix Polygoni Multiflori, Radix Glycyrrhizae, Poria, Radix Bupleuri, Radix Paeoniae, Rhizoma Chuanxiong, the Rhizoma Atractylodis Macrocephalae, the Radix Astragali, Flos Chrysanthemi, Herba Artemisiae Scopariae, Radix Angelicae Sinensis, Radix Puerariae, Folium Nelumbinis, Herba Taxilli, Semen Persicae, Radix Rehmanniae, Radix Codonopsis, Rhizoma Curcumae Longae, Radix Ginseng, Herb Gynostemmae Pentaphylli, Fructus Lycii, Ganoderma, Folium Acanthopanacis Senticosi, Fructus Ligustri Lucidi, Cordyceps, Radix Achyranthis Bidentatae, the Cortex Eucommiae, Radix Oenotherae erythrosepalae, Pollen Typhae, Flos Carthami, Radix Notoginseng, Myrrha, Folium Ginkgo, Fructus Hippophae, Rhizoma Polygoni Cuspidati, Pericarpium Citri Reticulatae, the Rhizoma Pinelliae, Radix Rhapontici, Rhizoma Coptidis, Radix Scutellariae, Ramulus Euonymi, Pinus densiflora leaf, Fructus Cannabis, Folium Kaki, pollen, Herba Portulacae, Semen sojae atricolor, Bulbus Allii, Folium Camelliae sinensis, Thallus Laminariae (Thallus Eckloniae), Testa oryzae, Lentinus Edodes, Thallus Porphyrae, Herba bromi japonici, Bulbus Allii Cepae, Radix Platycodonis, Bulbus Allii Macrostemonis, Hirudo, Fel Ursi, Auricularia etc.Be that master's Chinese medicine has: Fructus Crataegi, Rhizoma Alismatis, Radix Polygoni Multiflori, Radix Bupleuri, Rhizoma Chuanxiong, Radix Angelicae Sinensis, Folium Nelumbinis, Radix Codonopsis, Radix Ginseng, Ganoderma, Folium Acanthopanacis Senticosi, Radix Achyranthis Bidentatae, Pollen Typhae, Fructus Hippophae, Pericarpium Citri Reticulatae, the Rhizoma Pinelliae, Radix Rhapontici, Semen sojae atricolor, Bulbus Allii Macrostemonis etc. wherein with cholesterol reducing (TC).Triglyceride lowering (TG) is that main Chinese medicine has: Radix Glycyrrhizae, Folium Acanthopanacis Senticosi, Rhizoma Coptidis, Radix Scutellariae etc.The Chinese medicine that falls TC, TG has: Radix Et Rhizoma Rhei, Semen Cassiae, Radix Puerariae, Herba Taxilli, Rhizoma Curcumae Longae, Radix Ginseng, Herb Gynostemmae Pentaphylli, Fructus Lycii, Fructus Ligustri Lucidi, Cordyceps, Radix Oenotherae erythrosepalae, Radix Notoginseng, Folium Ginkgo, Rhizoma Polygoni Cuspidati, Herba Portulacae, Bulbus Allii, Folium Camelliae sinensis, Hirudo, Fel Ursi, Auricularia etc.Its effect for reducing fat is in the majority with serum cholesterol-lowering; Its effect belong to invigorating the liver and kidney, invigorating the spleen and benefiting QI, blood circulation promoting and blood stasis dispelling more and help digestion, expectorant, diuretic etc.
The research of 2 Chinese medicine blood fat reducing effective ingredient
Find have the various active composition to have good effect for reducing fat through a large amount of experimentatioies and clinical investigation, existing drug effect and mechanism of action with regard to multiple blood fat reducing active component summarized as follows.
Saponin component: the effective ingredient with effect for reducing fat is in the majority with saponins.The natural drug that is the master with this constituents effect for reducing fat has: Herb Gynostemmae Pentaphylli, Radix Ginseng, Radix Bupleuri, Folium Notoginseng, Folium Acanthopanacis Senticosi etc., they have remarkable reduction effect to TC, TG and the LDL of hyperlipemia animal.
Anthraquinone component: extensively be present in the natural drug, and in the majority with Polygonaceae, leguminous plant, mainly represent medicine to have: Radix Et Rhizoma Rhei, Radix Polygoni Multiflori, Rhizoma Polygoni Cuspidati etc.
Flavones ingredient: its physiologically active is various, and blood fat reducing mainly realizes through antioxidation.Represent in the plant, Fructus Crataegi is the conventional Chinese medicine material, and mainly containing hyperin is main flavonoid organic acid, can obviously reduce mice serum TC and rising HDL/TC; Buckwheat total flavone, Fructus Hippophae flavone, Folium Ginkgo flavone all can suppress the rising of hyperlipemia animal TC, TG, and hyperlipemia is had significant prevention and therapeutical effect.
The alkaloids composition: alkaloid is to have one type of maximum chemical constituents in the natural drug, has intensive physiologically active.As utilizing lotus leaf alkaloid preparation feed hyperlipemia in mice, the content of its serum TC has obvious reduction.
Volatile oil and fatty oils: volatile oil is one type of mixture of mainly being made up of terpenoid, low-molecular-weight fat family and aromatic compound.All belong to this type of like Oleum Hippophae, microula oil, Radix Oenotherae erythrosepalae oil, Chinese Oleum Bulbus Allii and Fructus Cannabis oil.
Protein-based composition: refer to that mainly some reactive proteins, bioactive peptide and aminoacid etc. are rich in special bioactive material.Soybean protein, Rhizoma Dioscoreae esculentae mucin, Semen Cassiae protein all belong to this type of, and TC, TG and the LDL of hyperlipidemia rats are significantly reduced.Because contain the component that combines with bile acid in this proteinoid, thereby suppress absorption of cholesterol and deposition in vivo.
The active polysaccharide class: blood fat reducing mechanism is with seemingly protein-based.Contained glucomannoglycan in LBP, Cordyceps militaris polysaccharide, laminarin, ulva polysaccharide and the Rhizoma amorphophalli can significantly reduce TC, TG level in the hyperlipemia animal blood fat, improves the HDL/TC value simultaneously.
Unsaturated fatty acid: it can combine with cholesterol to generate fusing point low, be easy to running, metabolism and excretory ester, change in the cholesterol body and distribute, reduce fat ester deposition in the blood vessel wall.
Polyphenol: blood fat reducing mechanism and flavonoid are seemingly also realized through antioxidation and removing free radical.
Other constituents: except above-mentioned blood fat reducing active component, also have the minority steroid, chemical compounds such as terpenoid.As: contained phytosterol can reduce absorption of cholesterol in the Semen phaseoli radiati.The Rhizoma Alismatis master contains triterpenoid compound, can reduce serum TC, TG, the rising serum hdl.
In sum, the Chinese medicine blood fat reducing is a kind of simple and efficient, efficient ways.In Chinese traditional medicines depressing lipid, some be medicine be again food, like Fructus Crataegi, Fructus Lycii, Folium Nelumbinis, Flos Chrysanthemi, Bulbus Allii, Thallus Laminariae (Thallus Eckloniae), Herba Portulacae etc., this for the exploitation fat-reducing food created good condition.But still have some problem demanding prompt solutions at present, like Chinese medicine in that to prevent and treat aspect the hyperlipemia low-level repetition phenomenon serious; Clinical experiment lacks the theoretical foundation of modern science mostly; The research short-term effect is far more than long-term effect; Do not carry out the research of profound active chemical in a lot of reports.So not only will carry out the screening of active drug from now on, and should in depth study pharmacology and chemical composition of Chinese materia medica, according to the structure activity relationship good prescription of lipid-lowering effect of drawing up out, so that be human health service better.At present, seek new Chinese traditional medicines depressing lipid, the Chinese medicine of having found is estimated becomes an important research direction.
Acute toxicity
Stigma Maydis flavone chmice acute toxicity toxicity is too low, can't record LD
50, maximum tolerated dose is 16g/kg.
Summary of the invention
The present invention provides the application of a kind of Stigma Maydis flavone in the medicine of treatment hyperlipidemia and fat-reducing, and purpose is to provide a kind of novel blood fat reducing, slimming medicine, satisfies the needs of people to such medicine.
The method that Stigma Maydis flavone of the present invention can be taked to put down in writing in conventional method or the document obtains, and is preferably to comprise the following steps:
1, technological process
Stigma Maydis → get the raw materials ready → defat → water carries → concentrates → decolour → alcohol analyses → supernatant → recovery ethanol → chromatography → drying → Stigma Maydis flavone
2, production control parameter and concrete operations
1. get the raw materials ready: choose free from insect pests, do not have go mouldy, fresh Stigma Maydis is raw material, clean, dry, pulverize, cross 40 mesh sieves, for use.
2. defat: the Stigma Maydis powder adds 95% ethanol, and soaked overnight is with the weeding of grease solubility impurity.Vacuum filtration, filter cake reuse same concentrations Ethanol Treatment is once, and is for use; Reclaim ethanol, recycle.
3. water is carried: the medical material powder after the defat is dropped in the round-bottomed flask, add 10～20 times of amounts deionized water, stir, soak 24h after, in 80～90 ℃ of heating 30min, extract 3 times, the united extraction clear liquid, for use.
4. concentrate: concentrating in the lixiviate stillness of night input rotary evaporation in vacuo appearance, decompression, cryoconcentration to feed liquid original volume about 1/5 get concentrated solution, and be for use.
5. decolouring: drop into concentrated solution in the round-bottomed flask, under agitation add a small amount of concentration and be 20～30% hydrogen peroxide, mix homogeneously, heating, bleaching time 2h, 50 ℃ of heating-up temperatures, filtered while hot, destaining solution, heating boils off residual hydrogen peroxide, and is for use.
6. alcohol is analysed: drop into destaining solution in the beaker, adds 95% an amount of ethanol, make that concentration of alcohol is 70%～75% (V/V) in the feed liquid, stir, mix homogeneously, be still in the refrigerator, alcohol is analysed and is spent the night, and liquid phase recovery ethanol is collected in centrifugalize, and is for use.
7. chromatography: reclaiming AB-8 macroporous adsorbent resin on the ethanol concentrated solution, after the washing, the different concentration ethanol eluting is collected 60%-70% ethanol part, reclaims ethanol, and is for use.
8. dry: to the vacuum drying baker, drying is crossed 100 mesh sieves, gets the Stigma Maydis flavone Stigma Maydis ethanol part.
9. with rutin make reference substance, aluminum chloride methanol makees developer and testing sample solution shakes up, placed 10 minutes, be solvent blank with 60% ethanol, at 272 blood ultraviolet determination absorbances, in the substitution regression equation, calculate percentage composition.Requiring purity is being qualified more than 50%.
Medicine of the present invention can adopt the conventional method of Chinese medicine preparation to be prepared into any conventional oral preparations, external preparation.
The beneficial effect of patent of the present invention
1, it is simple to use water as the method for distilling of solvent extraction flavone, concentrates the back ethanol precipitation, and the deposition part is a polysaccharide, and supernatant is the flavone part.Corn silk polysaccharide and flavone are extracted simultaneously energy savings.
2, adopt AB-8 that crude flavonoid powder is made with extra care, the purity of flavone is brought up to more than 50% by about 5%.And assay is carried out methodology examine or check, RSD is all less than 2%.
3, the new purposes Stigma Maydis present stage that increases plant resources is thrown away as garbage in vain, and is also very few to its research as medicine resource, belongs to the untapped kind of utilizing.With Stigma Maydis total flavones be developed to blood fat reducing, slimming preparation is to the utilization of Chinese herbal medicine resources effective.
4, driving development of regional economy Jilin Province is one of main producing region of corn, can produce 900000 tons of Stigma Maydis per year, and present stage is also effectively utilized, because hypoglycemic clinical drug demand is bigger, will drive the regional industrial expanding economy.
The medicine Stigma Maydis flavone effect for reducing blood fat that the present invention processes significantly and antiobesity action arranged.Taking dose can be according to the mode of taking, and patient's age and body weight and the degree that is in a bad way change with other similar factor, and oral dose is: 0.45-3.0g/ day people, divide and take for one～three time every day.
The specific embodiment
Further verify the present invention through pharmacodynamic experiment below.
First's Stigma Maydis total flavones is to the influence of normal rat fat
1. material and method
1.1 main agents, medicine and instrument
Stigma Maydis flavone Jilin Institute of Chemical Technology Stigma Maydis research and development group provides (content is 54%)
Chemical Industry Science Co., Ltd 20070416 is analysed in pentobarbital sodium Shanghai clearly
Normal saline Jilin Cornell pharmaceutical Co. Ltd 20070515
The digital display thermostat water bath state China HH-2 of Electrical Appliances Co., Ltd type
The low speed autobalancing centrifuge Beijing medical machine LDZ5-2 of factory type
The BS-600L of electronic balance Shanghai Yousheng Balance Co., Ltd. type
1.2 laboratory animal
The Wistar rat, male, 40, (Jilin University's Experimental Animal Center provides the quality certification number to body weight 120～160g: the moving word 10-5112 of doctor).
1.3 medicine preparation and animal divide into groups
1.3.1 medicine preparation
The Stigma Maydis flavone is made into 0.4%, 0.8% and 1.6% solution with distilled water.
1.3.2 animal divides into groups and dosage regimen
40 rats are divided into 4 groups at random by body weight, and 10 every group, gastric infusion, once a day, continuous 28d.
Blank control group distilled water 10mL/kg
Low dose group Stigma Maydis flavone 40mg/kg
Middle dose groups Stigma Maydis flavone 80mg/kg
High dose group Stigma Maydis flavone 160m/kg
1.4 experimental technique
During the administration, every 7d writes down the body weight of every group every rat, amount of drinking water and food ration.Before the last administration; Water 14h is can't help in each treated animal fasting; 30min after the last administration, with 3% pentobarbital sodium 30mg/kg intraperitoneal anesthesia, ventral aorta is taken a blood sample with animal; Separation of serum, the content of mensuration cholesterol in serum, triglyceride, high density lipoprotein, low density lipoprotein, LDL and free fatty.
2. statistical method
All results all adopt Excel to carry out group difference property t check, and the result representes with means standard deviation
.
3. result
3.1 the Stigma Maydis flavone is to the influence of normal rat amount of drinking water, food ration and body weight
Compare with blank control group, the middle and high dosage of Stigma Maydis flavone can reduce the amount of drinking water and the food ration of 21d and 28d normal rat, p＜0.05 or p＜0.01; See Tab.1 and Tab.2; The middle and high dosage of Stigma Maydis flavone can obviously reduce the body weight of normal rat, and Tab.3 is seen in p＜0.05 or p＜0.01.
Tab.1 Stigma Maydis flavone is to the influence (
n=10) of normal rat amount of drinking water
Compare with blank control group,
*P＜0.05,
*P＜0.01.
Tab.2 Stigma Maydis flavone is to the influence (
n=10) of normal rat food ration
Compare with blank control group,
*P＜0.05,
*P＜0.01.
Tab.3 Stigma Maydis flavone is to the influence (
n=10) of normal rat body weight
Compare with blank control group,
*P＜0.05,
*P＜0.01.
3.2 the Stigma Maydis flavone is to the influence of normal rat fat
Compare with blank control group; The Stigma Maydis flavone has the trend that reduces serum cholesterol content, reduces serum triglycerides content, reduces serum high-density LP content, increases serum low-density LP content and reduce the free serum content of fatty acid to normal rat; But not statistically significant is seen Tab.4.
Tab.4 Stigma Maydis flavone is to the influence (
n=10) of normal rat fat
4. conclusion
1. the Stigma Maydis flavone can reduce the amount of drinking water and the food ration of normal rat
2. the Stigma Maydis flavone has the reduction effect to the body weight of normal rat
3. the Stigma Maydis flavone has the trend of reduction to normal Serum TC, TG, LDL-c and FFA
In sum, the Stigma Maydis flavone can obviously reduce the body weight of normal rat under the situation that reduces amount of drinking water and food ration; The trend that normal rat is had blood fat reducing.
Second portion Stigma Maydis flavone is to the influence of hyperlipidemia rats blood fat
1. material and method
1.1 main agents, medicine and instrument
Stigma Maydis flavone Jilin Institute of Chemical Technology Stigma Maydis research and development group provides (content is 54%)
Zhaodong, lovastatin Heilungkiang Hua Fu pharmaceutcal corporation, Ltd 20091101
Propylthiouracil Shanghai Zhaohui Pharmaceutical Co., Ltd. 20090604
Cholesterol Tianjin recovery fine chemistry industry institute 20090630
Sodium cholate China favour generation biochemical reagents company limited 20050830
Chemical Industry Science Co., Ltd 20070416 is analysed in pentobarbital sodium Shanghai clearly
The C8000 of automatic clinical chemistry analyzer U.S. Abbott Laboratories type
The digital display thermostat water bath state China HH-2 of Electrical Appliances Co., Ltd type
The low speed centrifuge Beijing medical machine LDZ5-2 of factory type
The BS-600L of electronic balance Shanghai Yousheng Balance Co., Ltd. type
1.2 laboratory animal
The Wistar rat, male, 80, (Jilin University's Experimental Animal Center provides the quality certification number to body weight 120～160g: the moving word 10-5112 of doctor).
1.3 medicine preparation and animal divide into groups
1.3.1 high lipid food prescription
2.0% cholesterol, 0.2% propylthiouracil, 0.3% sodium cholate, 7.5% Adeps Sus domestica, 2..0% egg yolk, 88.0% normal feedstuff
1.3.2 medicine preparation
The Stigma Maydis flavone is made into 0.4%, 0.8% and 1.6% solution with distilled water; Lovastatin is made into 0.3% suspension with 0.5% sodium carboxymethyl cellulose.
1.3.3 animal divides into groups and dosage regimen
80 of Wistar male rats, body weight 120～160g is divided into blank control group (10) and high lipid food group (80) with it earlier; The blank control group normal diet; The high lipid food group is given high lipid food, feeds 21d continuously, gets blood; Separation of serum, the content (water 14h is can't help in fasting before getting blood) of cholesterol, triglyceride, high density lipoprotein, low density lipoprotein, LDL and free fatty in the mensuration high lipid food group rat blood serum.Choose the higher rat of blood fat, the high lipid food group is divided into 5 groups again according to blood lipid level, that is: model group, positive drug group, the basic, normal, high dose groups of Stigma Maydis flavone, 10 every group, gastric infusion, once a day, 28d continuously.
Blank control group normal diet distilled water 10mL/kg
Model group high lipid food distilled water 10mL/kg
Lovastatin group high lipid food lovastatin 30mg/kg
Low dose group high lipid food Stigma Maydis flavone 40mg/kg
Middle dose groups high lipid food Stigma Maydis flavone 80mg/kg
High dose group high lipid food Stigma Maydis flavone 160mg/kg
1.4 experimental technique
During the administration, every 7d writes down the body weight of every group every rat, water uptake and food-intake.Before the last administration; Water 14h is can't help in each treated animal fasting, 30min after the last administration, with animal with 3% pentobarbital sodium 30mg/kg intraperitoneal anesthesia; The ventral aorta blood sampling, the content of mensuration cholesterol in serum, triglyceride, high density lipoprotein, low density lipoprotein, LDL and free fatty.
2. statistical method
All results all adopt Excel to carry out group difference property t check, and the result is with means standard deviation (x ± s) expression.
3. result
3.1 the Stigma Maydis flavone is to the influence of hyperlipidemia rats amount of drinking water, food ration and body weight
Compare with model group, the middle and high dosage of Stigma Maydis flavone can reduce the amount of drinking water and the food ration of 28d hyperlipidemia rats, and Tab.5 and Tab.6 are seen in p＜0.05 or p＜0.01; Compare with model group, the basic, normal, high dosage of corn silk polysaccharide can obviously reduce the body weight of hyperlipidemia rat, and Tab.7 is seen in p＜0.05.
Tab.5 Stigma Maydis flavone is to the influence (
n=10) of hyperlipidemia rats amount of drinking water
Compare with model group,
*P＜0.05,
*P＜0.01.
Tab.6 Stigma Maydis flavone is to the influence (
n=10) of hyperlipidemia rats food ration
Compare with model group,
*P＜0.05,
*P＜0.01.
Tab.7 Stigma Maydis flavone is to the influence (
n=10) of hyperlipidemia rats body weight
Compare with model group,
*P＜0.05,
*P＜0.01.
3.2 the Stigma Maydis flavone is to the influence of hyperlipidemia rats blood fat
Compare with blank control group, model group TC and LDL-c significantly raise, and p＜0.01 explains that blood lipid metabolism is disorderly, hyperlipemia model modeling success; Compare with model group; Positive drug can reduce hyperlipemia rat serum TC, TG, LDL-c level; P＜0.05 or p＜0.01, the middle and high dose groups of Stigma Maydis flavone can reduce type 2 diabetes mellitus Serum TC, TG, LDL-c and FFA level, and Tab.8 is seen in p＜0.05 or p＜0.01.
Tab.8 Stigma Maydis flavone is to the influence (
n=10) of hyperlipidemia rats blood fat
Compare △ P＜0.05, △ △ P＜0.01 with blank control group; Compare with model group,
*P＜0.05,
*P＜0.01.
4. conclusion
1. the Stigma Maydis flavone can reduce the amount of drinking water and the food ration of hyperlipidemia rats
2. the Stigma Maydis flavone can obviously reduce the body weight of hyperlipidemia rats.
3. the Stigma Maydis flavone can obviously reduce TC, TG, LDL-c and the FFA level of hyperlipidemia rats serum
In sum, the Stigma Maydis flavone can obviously reduce the body weight of hyperlipidemia rats under the situation that reduces hyperlipidemia rats amount of drinking water and food ration, have the effect that reduces the hyperlipidemia rats blood fat.
Third part Stigma Maydis flavone is to the influence of type diabetes rat blood fat
1. material and method
1.1 main agents, medicine and instrument
Stigma Maydis flavone Jilin Institute of Chemical Technology Stigma Maydis research and development group provides (content is 54%)
Metformin Jiangsu Heng Shan pharmaceutcal corporation, Ltd 20091201
Propylthiouracil Shanghai Zhaohui Pharmaceutical Co., Ltd. 20090604
Cholesterol Tianjin recovery fine chemistry industry institute 20090630
Sodium cholate China favour generation biochemical reagents company limited 20050830
Chemical Industry Science Co., Ltd 20070416 is analysed in pentobarbital sodium Shanghai clearly
The C8000 of automatic clinical chemistry analyzer U.S. Abbott Laboratories type
The digital display thermostat water bath state China HH-2 of Electrical Appliances Co., Ltd type
The low speed centrifuge Beijing medical machine LDZ5-2 of factory type
The BS-600L of electronic balance Shanghai Yousheng Balance Co., Ltd. type
The SXT-1 of blood glucose meter Changsha Sannuo Biology Sensing Technology Co., Ltd type
1.2 laboratory animal
The Wistar rat, male, 80, (Jilin University's Experimental Animal Center provides the quality certification number to body weight 120～160g: the moving word 10-5112 of doctor).
1.3 medicine preparation and animal divide into groups
1.3.1 high lipid food prescription
2.0% cholesterol, 0.2% propylthiouracil, 0.3% sodium cholate, 7.5% Adeps Sus domestica, 2.0% egg yolk, 88.0% normal feedstuff
1.3.2 medicine preparation
The Stigma Maydis flavone is made into 0.4%, 0.8% and 1.6% solution with distilled water; Metformin is made into 0.4% suspension with 0.5% sodium carboxymethyl cellulose; Alloxan is mixed with 12% solution with normal saline, and is existing with join at present.
1.3.3 animal divides into groups and dosage regimen
80 rats are divided into blank control group (10) and high lipid food group (70) earlier, the blank control group normal diet, and the high lipid food group gives high lipid food.Feed 21d continuously; Get blood; Separation of serum is measured the content and the blood glucose value of cholesterol, triglyceride, high density lipoprotein, low density lipoprotein, LDL, free fatty in the high lipid food group rat blood serum and is measured blank control group blood glucose value (water 14h is can't help in equal fasting before getting blood).Choose the higher rat of blood fat in the high lipid food group, the lumbar injection alloxan, 120mg/kg carries out modeling to it.After three days, measure the fasting blood sugar of modeling rat, choose the rat of blood glucose value＞11.1mmol/L; According to blood glucose, blood fat and body weight the modeling rat is divided into 5 groups again; That is: model group, positive drug group, the basic, normal, high dose groups of Stigma Maydis flavone, 10 every group, gastric infusion; Once a day, continuous 28d.
Blank control group normal diet distilled water 10mL/kg
Model group high lipid food distilled water 10mL/kg
Metformin group high lipid food metformin 40mg/kg
Low dose group high lipid food Stigma Maydis flavone 40mg/kg
Middle dose groups high lipid food Stigma Maydis flavone 80mg/kg
High dose group high lipid food Stigma Maydis flavone 160mg/kg
1.4 experimental technique
During the administration, write down the body weight of every group of rat, water uptake and food-intake.In 7d, 14d and 21d, behind the administration 30min, the tail vein is got blood, with the blood glucose value of measuring every group of rat (getting not only water 14h of blood fasting before).Before the last administration; Water 14h is can't help in each treated animal fasting; 30min after the last administration measures the blood glucose value of every group of rat, with 3% pentobarbital sodium 30mg/kg intraperitoneal anesthesia; The ventral aorta blood sampling, the content of mensuration cholesterol in serum, triglyceride, high density lipoprotein, low density lipoprotein, LDL and free fatty.
2. statistical method
All results all adopt Excel to carry out group difference property t check, and the result representes with means standard deviation (
n=10).
3. result
3.1 the Stigma Maydis flavone is to the influence of type 2 diabetes mellitus rat blood sugar
Compare with blank control group, model group 7d, 14d, 21d, 28d blood glucose value all significantly raise, and hyperglycemia model modeling success is explained in p＜0.01; Compare with model group, positive drug can reduce the blood glucose value of 21d and 28d2 type diabetes rat, p＜0.05, and each organizes the trend that the blood glucose of type 2 diabetes mellitus rat is had reduction the Stigma Maydis flavone, but not statistically significant is seen Tab.9.
Tab.9 Stigma Maydis flavone is to the influence (
n=10) of type 2 diabetes mellitus rat blood sugar
Compare △ P＜0.05, △ △ P＜0.01 with blank control group; Compare with model group,
*P＜0.05,
*P＜0.01.
3.2 the Stigma Maydis flavone is to the influence of type 2 diabetes mellitus rat fat
Compare with blank control group, model group TC, TG, LDL-c significantly raise, and p＜0.01 explains that blood lipid metabolism is disorderly, type 2 diabetes mellitus model modeling success; Compare with model group; Positive drug can reduce type 2 diabetes mellitus Serum TC, TG, LDL-c level; P＜0.05 or p＜0.01; The middle and high dose groups of Stigma Maydis flavone can reduce type 2 diabetes mellitus Serum TC and FFA level, and the basic, normal, high dose groups of Stigma Maydis flavone can reduce type 2 diabetes mellitus rat blood serum TG and LDL-c level, and Tab.10 is seen in p＜0.05 or p＜0.01.
Tab.10 Stigma Maydis flavone is to the influence (
n=10) of type 2 diabetes mellitus rat fat
Compare △ P＜0.05, △ △ P＜0.01 with blank control group; Compare with model group,
*P＜0.05,
*P＜0.01.
4. conclusion
1. the Stigma Maydis flavone has the trend of reduction to the blood glucose of type 2 diabetes mellitus rat
2. the Stigma Maydis flavone reduces TC, TG, LDL-c and the FFA level of type 2 diabetes mellitus rat
In sum, the blood glucose of Stigma Maydis flavone type 2 diabetes mellitus rat has the trend of reduction, has the effect of the blood fat that reduces the type 2 diabetes mellitus rat.
Method for preparing of the present invention:
Embodiment 1
1. get the raw materials ready: choose free from insect pests, do not have go mouldy, fresh blue or green Stigma Maydis is raw material, clean, dry, pulverize, cross 40 mesh sieves, for use.
2. defat: the Stigma Maydis powder adds 95% ethanol, and soaked overnight is with the weeding of grease solubility impurity.Vacuum filtration, filter cake reuse same concentrations Ethanol Treatment is once, and is for use; Reclaim ethanol, recycle.
3. water is carried: the medical material powder after the defat is dropped in the round-bottomed flask, add 10～20 times of amounts deionized water, stir, soak 24h after, in 80～90 ℃ of heating 30min, extract 3 times, merge extractive liquid,, for use.
4. concentrate: concentrate extracting to drop into night in the rotary evaporation in vacuo appearance, decompression, cryoconcentration to feed liquid original volume about 1/5 get concentrated solution, and be for use.
5. decolouring: drop into concentrated solution in the round-bottomed flask, under agitation add a small amount of concentration and be 20～30% hydrogen peroxide, mix homogeneously, heating, bleaching time 2h, 50 ℃ of heating-up temperatures, filtered while hot, destaining solution, heating boils off residual hydrogen peroxide, and is for use.
6. alcohol is analysed: drop into destaining solution in the beaker, adds 95% an amount of ethanol, make that concentration of alcohol is 70%～75% (V/V) in the feed liquid, stir, mix homogeneously, be still in the refrigerator, alcohol is analysed and is spent the night, and liquid phase recovery ethanol is collected in centrifugalize, and is for use.
7. chromatography: reclaiming AB-8 macroporous adsorbent resin on the ethanol concentrated solution, after the washing, the different concentration ethanol eluting is collected 60%-70% ethanol part, reclaims ethanol, and is for use.
8. dry: to the vacuum drying baker, drying is crossed 100 mesh sieves, gets the Stigma Maydis flavone Stigma Maydis ethanol part.
9. with rutin make reference substance, aluminum chloride methanol makees developer and testing sample solution shakes up, placed 10 minutes, be solvent blank with 60% ethanol, at 272nm ultraviolet determination absorbance, in the substitution regression equation, calculate percentage composition.Requiring purity is being qualified more than 50%.
Embodiment 2
1. get the raw materials ready: choose free from insect pests, do not have go mouldy, fresh blue or green Stigma Maydis is raw material, clean, dry, pulverize, cross 40 mesh sieves, for use.
2. defat: the Stigma Maydis powder adds 95% ethanol, and soaked overnight is with the weeding of grease solubility impurity.Vacuum filtration, filter cake reuse same concentrations Ethanol Treatment is once, and is for use; Reclaim ethanol, recycle.
3. alcohol extraction: the medical material powder after the defat is dropped in the round-bottomed flask, add the ethanol of 10～20 times of amount 60%-70%, stir, behind the immersion 24h,, extract 3 times in 70～80 ℃ of heating 30min, merge extractive liquid,, for use.
4. concentrate: concentrate extracting to drop into night in the rotary evaporation in vacuo appearance, decompression, cryoconcentration to feed liquid original volume about 1/5 get concentrated solution, and be for use.
5. decolouring: drop into concentrated solution in the round-bottomed flask, under agitation add a small amount of concentration and be 20～30% hydrogen peroxide, mix homogeneously, heating, bleaching time 2h, 50 ℃ of heating-up temperatures, filtered while hot, destaining solution, heating boils off residual hydrogen peroxide, and is for use.
6. chromatography: AB-8 macroporous adsorbent resin on the destaining solution, after the washing, the different concentration ethanol eluting is collected 60%-70% ethanol part, reclaims ethanol, and is for use.
7. dry: partly put Stigma Maydis ethanol in the vacuum drying oven, drying is crossed 100 mesh sieves, gets the Stigma Maydis flavone.
8. with rutin make reference substance, aluminum chloride methanol makees developer and testing sample solution shakes up, placed 10 minutes, be solvent blank with 60% ethanol, at 272nm ultraviolet determination absorbance, in the substitution regression equation, calculate percentage composition.Requiring purity is being qualified more than 50%.
Embodiment 3
The technical study of the refining Stigma Maydis crude flavonoid powder of AB-8
The Stigma Maydis water extract is concentrated into 1/5 of original solution volume, adds 95% ethanol, make solution concentration reach 70-80% by amount of calculation; Left standstill 24 hours; Solution is collected in centrifugal back, promptly gets Stigma Maydis water extract-alcohol precipitation supernatant, and its concentrated final vacuum evaporate to dryness is got drying sample powder (crude flavonoid); Take by weighing the above-mentioned powder of 40mg, place the 100mL volumetric flask, it is an amount of to add 70% ethanol, ultrasonic make it the dissolving, put cold and with 70% ethanol dilution to scale, shake up, promptly get need testing solution.
In order to reduce experimental cost, raise the efficiency, make process conditions be able to further optimization, exploring experiment gained optimum condition with single factor is benchmark, dwindles the horizontal extent of corresponding factor, selected orthogonal experiment three factors three levels:
Tab.11 orthogonal experiment factor and design levels table
Tab.12L9 (3
4) Orthogonal experiment results
Can draw through a series of orthogonal experiment: with AB-8 type macroporous adsorbent resin Stigma Maydis total flavones is made with extra care, the optimum process condition that draws is: eluant flow velocity 3.6mL/min, eluant pH value 5.61, eluant strength 70%; Adopt optimized process conditions, the purity of total flavones brings up to 58.32% by 14.24%, and yield has reached 69.35%.Explain that the AB-8 resin has preferably specificity conciliate absorption property to Stigma Maydis total flavones, concentration effect is good, has easy and simple to handlely, and the products obtained therefrom purity and the rate of transform are higher, are easy to realize suitability for industrialized production.
Embodiment 4
The preparation and the quality control of Stigma Maydis flavone capsule for descend of blood fat
1. raw material is pressed the method extraction of embodiment 1 or 2, and the method for embodiment 3 is refining, puts 70 ℃ of vacuum dryings, pulverizes, and crosses No. 7 standard screens, and is subsequent use.
2. process hard capsule above-mentioned medicated powder adding calcium carbonate and starch are stirred, sieve.Add ethanol adjustment humidity in case of necessity,, in 1 or No. 2 capsule of packing into, process 1000 altogether in 60 ℃～70 ℃ oven dry.Conventional sterilization packs every capsule 0.03～0.2g that heavily contains the Stigma Maydis flavone behind the granulate.Every day three times, each 3～5 balls.
3. quality control outward appearance, moisture, content uniformity, disintegration, sanitary standard all meet the requirements.
4. the Stigma Maydis flavone capsule has that technology is simple, steady quality, purity are high, dosage accurately, determined curative effect, taking convenience is convenient to advantages such as identification, storage and transportation.
Claims (2)
1. the application of Stigma Maydis flavone in preparation treatment hyperlipidemia medicine is characterized in that the Stigma Maydis flavone is obtained by following method for preparing:
1. get the raw materials ready: choose free from insect pests, do not have go mouldy, fresh Stigma Maydis is raw material, clean, dry, pulverize, cross 40 mesh sieves, for use;
2. defat: the Stigma Maydis powder adds 95% ethanol, and soaked overnight is with the weeding of grease solubility impurity.Vacuum filtration, filter cake reuse same concentrations Ethanol Treatment is once, and is for use; Reclaim ethanol, recycle;
3. water is carried: the medical material powder after the defat is dropped in the round-bottomed flask, add 10～20 times of amounts deionized water, stir, soak 24h after, in 80～90 ℃ of heating 30min, extract 3 times, merge extractive liquid,, for use;
4. concentrate: concentrate carrying dropping into night in the rotary evaporation in vacuo appearance, decompression, cryoconcentration to feed liquid original volume about 1/5 get concentrated solution, and be for use;
5. decolouring: drop into concentrated solution in the round-bottomed flask, under agitation add a small amount of concentration and be 20～30% hydrogen peroxide, mix homogeneously, heating, bleaching time 2h, 50 ℃ of heating-up temperatures, filtered while hot, destaining solution, heating boils off residual hydrogen peroxide, and is for use;
6. alcohol is analysed: drop into destaining solution in the beaker, adds 95% an amount of ethanol, make that concentration of alcohol is 70%～75% (V/V) in the feed liquid, stir, mix homogeneously, be still in the refrigerator, alcohol is analysed and is spent the night, and liquid phase recovery ethanol is collected in centrifugalize, and is for use;
7. chromatography: reclaiming AB-8 macroporous adsorbent resin on the ethanol concentrated solution, after the washing, the different concentration ethanol eluting is collected 60%-70% ethanol part, reclaims ethanol, and is for use;
8. dry: to the vacuum drying baker, drying is crossed 100 mesh sieves, gets the Stigma Maydis flavone Stigma Maydis ethanol part;
9. with rutin make reference substance, aluminum chloride methanol makees developer and testing sample solution shakes up, placed 10 minutes, be solvent blank with 60% ethanol, at 272nm ultraviolet determination absorbance, in the substitution regression equation, calculate percentage composition.Requiring purity is being qualified more than 50%.
2. the application of Stigma Maydis flavone as claimed in claim 1 in preparation treatment slimming medicine.
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| CN102178873B (en) * | 2011-04-23 | 2012-11-28 | 王效山 | Chinese medicine weight-losing preparation comprising external preparation and internal preparation and preparation method thereof |
| CN102908538A (en) * | 2011-08-02 | 2013-02-06 | 苏州宝泽堂医药科技有限公司 | Extraction method of corn stigma total flavonoids |
| CN102908599A (en) * | 2012-06-19 | 2013-02-06 | 谢文龙 | Medicine capable of reducing blood sugar, blood fat and blood pressure |
| CN103652161A (en) * | 2013-11-18 | 2014-03-26 | 肖森林 | Corn stigma medlar tea |
| KR20150061115A (en) * | 2013-11-25 | 2015-06-04 | 가톨릭대학교 산학협력단 | Pharmaceutical composition comprising maysin for the prevention and treatment of obesity |
| CN104082374A (en) * | 2014-07-04 | 2014-10-08 | 贵州黄平靓鸥桑综合开发有限公司 | Biscuits capable of reducing blood fat |
| CN105011287A (en) * | 2015-08-14 | 2015-11-04 | 青岛浩大海洋保健食品有限公司 | Rose and sea functionality health drink with blood lipid regulating effect |
| CN106070887A (en) * | 2016-06-26 | 2016-11-09 | 贾恩助 | A kind of damp eliminating fat-eliminating tea |
| CN107183427A (en) * | 2017-05-17 | 2017-09-22 | 霍山延年居石斛科技有限公司 | A kind of stem of noble dendrobium juice and preparation method thereof |
| CN112956624A (en) * | 2021-03-17 | 2021-06-15 | 吉林化工学院 | Research on stability of corn stigma compound beverage |
| CN113243528B (en) * | 2021-04-26 | 2023-05-23 | 上海市农业科学院 | Corn silk extract capable of efficiently reducing blood sugar and blood fat and preparation method thereof |
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