CN103766901A - Application of andrographolide C to preparation of weight-losing food or medicine - Google Patents

Application of andrographolide C to preparation of weight-losing food or medicine Download PDF

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CN103766901A
CN103766901A CN201410009603.8A CN201410009603A CN103766901A CN 103766901 A CN103766901 A CN 103766901A CN 201410009603 A CN201410009603 A CN 201410009603A CN 103766901 A CN103766901 A CN 103766901A
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neoandrographolide
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andrographolide
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陈卫
郑晓冬
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Zhejiang University ZJU
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/38—Other non-alcoholic beverages
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/20—Agglomerating; Granulating; Tabletting
    • A23P10/28—Tabletting; Making food bars by compression of a dry powdered mixture
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
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    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K9/2004—Excipients; Inactive ingredients
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    • A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
    • A61K9/2059—Starch, including chemically or physically modified derivatives; Amylose; Amylopectin; Dextrin
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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Abstract

The invention discloses an application of andrographolide C to preparation of a weight-losing food or medicine and an application of a medicine composition composed of the andrographolide C which is used as an active component and a pharmaceutically-acceptable additive to preparation of the weight-losing food or medicine. According to the application, the technical prejudices in the prior art that andrographolide does not have the weight-losing effect are overcome and an application field of the andrographolide C is expanded. A cell experiment shows that the andrographolide C can inhibit the proliferation of 3T3-L1 preadipocytes through an oxidative stress way; the preadipocytes can be effectively prevented from being differentiated to fat cells and lipids in the fat cells are promoted to be decomposed. A mouse experiment shows that the andrographolide C has obvious effects on reducing the growth rate of body mass of a mouse, the food utilization rate and the in-vivo fat, and a dosage dependency relation is formed; the mouse blood glucose level and the serum cholesterol level can be effectively reduced, and the serum free fatty acid level and the oxidation resistance level are improved.

Description

A kind of Neoandrographolide is in the application of preparing in diet food or medicine
Technical field
The present invention relates to Chinese medicine applied technical field, be specifically related to a kind of Neoandrographolide in the application of preparing in diet food or medicine.
Background technology
Fat (obesity) refers to that obvious overweight and fat deposit is to a certain degree blocked up, is body fat, and especially triglycerides gathers a kind of state too much causing.It does not refer to that simple body weight increases, but the superfluous state of body fat tissue savings.In recent years, no matter be the puzzlement that developed country or developing country are all faced with obesity accelerated development, bring huge medical expense and burden on society thereupon.From using in 19th century, medicine treatment is fat to be started, and slimming medicines are quit listing because of safety issue in a large number, as sibutramine, fenfluramine, Isomeride, phenylpropanolamine, profit not just like that etc.In view of the popular disease of the more serious obesity in the whole world, and effectively prevention and the treatment measure of current shortage, seek safe and effective prevention and therapeutic strategy, be extremely important for reducing the obesity incidence of disease.
In recent years, be one of focus of studying of scientific worker from the fat preparation of intervening of cellular level research.Wherein, front adipocyte (3T3-L1) is fusiformis, and volume is little, and interstitial is few, in endochylema, does not drip containing triacylglycerol fat; Add derivant (IBMX, 3-methyl isophthalic acid-isobutyl group xanthine; Dexamethasone; Insulin; Referred to as MDI) after induction, 3T3-L1 is divided into ripe adipocyte, and interstitial is abundant, and a large amount of triacylglycerol fat drips and accumulates in around core, is typical mature fat cell form.It has become the most popular natural function factor and has intervened one of fat medicaments sifting model.
Herba Andrographitis (Andrographispaniculata (Burm.f.) Nees) has another name called Banlangen, cuts snakeweed, eel grass, Lan Helian, is Acanthaceae Herba Andrographitis platymiscium, is a kind of conventional Chinese herbal medicine, has in the Orient long history in therapeutic treatment.China's Herba Andrographitis aboundresources, mainly originates in Guangdong, Fujian, Guangxi, and existing Shaanxi, Hainan, Jiangsu, Shandong, zhejiang and other places also have plantation.It is reported, that herb has is antipyretic, resisting pathogenic microbes, anti-inflammatory, enhancing immunologic function, antifertility, antivenomous effect.Can be used for clinically treating common cold, tracheitis, cholecystitis, liver diseases, bleb, pneumonia in pertussis, pulmonary tuberculosis, lung abscess, hypertension, acute bacillary dysentery, gastroenteritis, the swollen scald of sore furuncle carbuncle and venomous snake bite.The conventional use of Herba Andrographitis and effect of pharmacology aspect are recently studies confirm that in a large number.
The active component of Herba Andrographitis mainly comprises diterpenoid-lactone, polyphenol and Flavonoid substances.But its physiologically active is mainly given the credit to its active ingredient diterpene ginkgolide andrographolide (Andrographolide, molecular formula C20H30O5) and derivative thereof.Andrographolide and derivative thereof are mainly present in leaf, can from plant extracts, separate and obtain its crystalline solid.Andrographolide or derivatives thereof has BA widely, and research shows that it has antiphlogistic antibacterial, antitumor, viral infection resisting, anti-cardiovascular disease activity, has the effects such as the immunity of raising, hepatic cholagogic in addition.
The patent documentation that is CN101125850A as publication number discloses the application of andrographolidume derivative in hypoglycemic and diabetes medicament; Publication number is that the patent documentation of CN102247356A discloses the purposes of andrographolide at prevention and treatment hyperlipidemia and atherosclerosis disease, and discloses method prepared by its pharmaceutical preparation.
Publication number is that the patent documentation of CN102784137A discloses a kind of antiadipositas drug compositions and application thereof; a kind of antiadipositas drug compositions and application thereof of protecting melbine and andrographolide is provided, and this invention shows that andrographolide has the humidification of the anti-obesity effect to melbine.Known by the table 1 in more specific embodiment, in the high fat diet of feeding mouse, in diet, sneak into andrographolide 100mg/kg, oral processing is after one week and three weeks, the body weight of mouse is compared with the high-fat mouse of feeding only, lose weight not obvious, illustrate that the andrographolide adopting in this patent is to the not effect of anti-obesity of mouse.
Summary of the invention
The present invention is based on the further investigation to andrographolide and derivative thereof, provide a kind of Neoandrographolide in the new purposes of preparing in diet food or medicine, the present invention has overcome and in prior art, has thought that andrographolide does not possess the technology prejudice of fat-reducing effect, has expanded the application of Neoandrographolide.
The invention discloses a kind of Neoandrographolide in the application of preparing in diet food or medicine.
The invention also discloses a kind of Neoandrographolide as the pharmaceutical composition of active component and pharmaceutically acceptable additives composition in the application of preparing in diet food or medicine.
The molecular formula of described Neoandrographolide is as follows:
Figure BDA0000454509570000031
As preferably, described Neoandrographolide separation and Extraction from Herba Andrographitis obtains, concrete preparation technology can the patent documentation that be CN101559088A referring to publication number in disclosed method.
Andrographolide the third element in the present invention also can be obtained or be used conventional method synthesize and make by commercial sources.
As preferably, described pharmaceutical composition is tablet, is made up of the raw material of following mass percent:
Figure BDA0000454509570000032
Described filler is one or more in starch, microcrystalline cellulose, lactose.
Described disintegrant is one or more in sodium carboxymethyl starch, Ac-Di-Sol, PVPP, low-substituted hydroxypropyl cellulose, dried starch.
Described lubricant is one or more in talcum powder, dolomol, superfine silica gel powder, hydrogenated vegetable oil.
Further preferably, described tablet is made up of the raw material of following mass percent:
Figure BDA0000454509570000033
Described diet food is beverage based food, and the recommendation consumption of Neoandrographolide is 50～150mg/ days.
Described slimming medicine can be any applicable regular dosage form, and to be grown up as example, the recommendation consumption of Neoandrographolide is 50～150mg/ days, is preferably 100～150mg/ days.
Prove by cell experiment: Neoandrographolide can suppress the front lipocyte proliferation of 3T3-L1 by oxidative stress approach, can effectively suppress front adipocyte to Adipocyte Differentiation, promotes adipocyte inner lipid to decompose.Neoandrographolide is acted on to adipocyte before 3T3-L1, and action time is respectively 24h, 48h and 72h, and cell half-inhibition concentration IC50 is respectively 16.4 μ g/mL, 13.0 μ g/mL, and 8.6 μ g/mL.Neoandrographolide processed group is compared with control group, and differentiation inhibiting rate reaches 18.5%; The inhibiting rate that lipid is generated reaches 33%; The facilitation that lipid is decomposed reaches 1.45 times; By the adipocyte of differentiation is carried out to vitality test, find that 10 μ g/mL dosage Neoandrographolides do not have toxicity to the adipocyte of differentiation.
Prove by mouse experiment: Neoandrographolide has obvious reducing effect to weight of mice rate, food utilization, body fat, and be dose-dependence; Mouse blood sugar level, serum cholesterol level be can effectively reduce, serum free fatty acid levels, antioxidant levels improved.
Compared with prior art, the present invention has the following advantages:
1, to found through experiments Neoandrographolide lower to the required concentration for the treatment of of obesity in the present invention, has no side effect.
2, Neoandrographolide has hepatic cholagogic, strengthens the BAs such as immunity, has not only guaranteed the validity of fat-reducing, and body health is also played to facilitation.
3, the present invention, for Neoandrographolide provides new medical application, has expanded a new application.
Accompanying drawing explanation
Fig. 1 is the impact (* p<0.05, * * p<0.01 with control group compared with) of variable concentrations Neoandrographolide on front lipocyte proliferation vigor;
Fig. 2 be Neoandrographolide to 3T3-L1 before the inhibitory action of Adipocyte Differentiation:
MDI is the differentiation group (* p<0.05, * * p<0.01 is compared with MDI) through derivant induction;
Fig. 3 is the impact (* p<0.05, * * p<0.01 is compared with MDI) of the 3T3-L1 cell viability of Neoandrographolide on differentiation;
Fig. 4 is the impact (* p<0.05, * * p<0.01 with MDI compared with) of Neoandrographolide on triglycerides accumulation in adipocyte;
Fig. 5 is the impact (* p<0.05, * * p<0.01 with MDI compared with) of Neoandrographolide on the effect of adipocyte solution fat;
Fig. 6 is each test group Mouse Weight increment (* p<0.05, * * p<0.01 is compared with HFD);
Fig. 7 is the impact (* p<0.05, * * p<0.01 with HFD compared with) of Neoandrographolide on C57BL/6 mouse food ration and food utilization;
Fig. 8 is the impact (* p<0.05, * * p<0.01 with HFD compared with) of Neoandrographolide on C57BL/6 mouse liver weight (A) and adipose tissue weight (B);
Fig. 9 is the impact (* p<0.05, * * p<0.01 with HFD compared with) of Neoandrographolide on C57BL/6 mouse's blood sugar content (A), serum total cholesterol content (B), serum triglyceride content (C), free fatty acid content (D);
Figure 10 is the impact (* p<0.05, * * p<0.01 with HFD compared with) of Neoandrographolide on C57BL/6 mouse adipose tissue form.
The specific embodiment
Below in conjunction with specific embodiment, the invention will be further described, and what below enumerate is only specific embodiments of the invention, but protection scope of the present invention is not limited in this:
Embodiment 1
Functional weight-reducing drinks (according to mass fraction) containing Neoandrographolide:
Neoandrographolide 0.0375%, citric acid 0.25%, honey 6%, Aspartame 1.5%, flavoring essence 0.05%, sodium carboxymethylcellulose 1%, water 91.20%.
According to above-mentioned formula, by 3.75 grams of Neoandrographolides, 25 grams of citric acids, 63.5 grams of honey, 150 grams of Aspartames, 5 grams of flavoring essences, 100 grams of sodium carboxymethylcelluloses, 9120g water mixes, and boils sterile filling beverage bottle and get final product.Every bottle of 200ml, 75 milligrams/bottle of Determination of Andrographolides.
Dosage for being grown up: take each 1 bottle every day 2 times.
Embodiment 2
Neoandrographolide tablet (according to mass fraction):
Neoandrographolide 50%, starch 17.3%, lactose 25%, crosslinked carboxymethyl fecula sodium 7%, dolomol 0.7%.
According to the mass fraction of above-mentioned formula, by 75 grams of Neoandrographolides, 26.25 grams of starch are dissolved in 150 grams of water, the dry Neoandrographolide microcapsule granule of making of spraying.By microcapsules grain and 37.5 grams of lactose, 10.5 grams, crosslinked carboxymethyl fecula sodium, 1.05 grams of mixing of dolomol, join in three-dimensional motion mixer, and incorporation time is 15-20 minute, obtains total mixture.Then direct tablet compressing obtains 1000 altogether, and every 0.15 gram, 75 milligrams/of Neoandrographolide content
Dosage for being grown up: take each 1 every day 2 times.
Fat-reducing effect test
Test one: observe Neoandrographolide on 3T3-L1 before the impact of lipocyte proliferation vigor, the action pathway of lipocyte proliferation before understanding it and suppressing.
Experiment material:
Adipocyte before 3T3-L1, purchased from Chinese Academy of Sciences's cell bank;
Neoandrographolide (purity >98%), purchased from National Institute for Food and Drugs Control.
Tetramethyl nitroblue (MTT), sough (DMSO) in dimethyl Asia, all purchased from Sigma company.
Experimental technique:
Cell is cultivated: in the time that adipocyte before 3T3-L1 grows to 80% left and right of culture dish area, can go down to posterity.Concrete grammar is as follows: inhale and abandon old culture medium, with PBS cleaning 2 times, add 1mL trypsase, put back to CO 2in incubator, hatch.After 2～5min, add 8mL fresh culture to stop digestion, softly piping and druming, sucks cell suspension in aseptic centrifuge tube the centrifugal 5min of 1200r/min.Supernatant discarded adds 5mL fresh culture in centrifuge tube, after suction pipe piping and druming evenly, is inoculated in the culture dish containing 8mL fresh culture, puts into CO 2incubator is cultivated.
Mtt assay detects cell viability: select the cell of exponential phase, use trypsinization cell monolayer, be made into individual cells suspension with fresh culture, with 5 × 10 4the density of individual/mL is inoculated in 96 orifice plates, every hole 100 μ L.Put 37 ℃ of incubators and cultivate after 24 hours, every hole adds andrographolide by concentration gradient, sets up blank simultaneously, processes respectively 3T3-L1 front adipocyte 24h, 48h and 72h.The centrifugal 5min of 1500r/min, discards solution in 96 orifice plates, and every hole adds 150 μ LPBS to clean 1 time, and centrifugal (1500r/min) 5min, discards PBS again.Every hole adds 100 μ L culture mediums, the MTT that is 0.5mg/mL containing final concentration, put incubator and hatch 4h, complete to crystallization, 3000r/min is centrifugal, abandons solution in 96 orifice plates, and every hole adds DMSO solution 150 μ L, after bluish violet crystallization is dissolved completely, use ELIASA to detect the absorbance value (OD value) at wavelength 490nm place.Be calculated as follows the impact of Neoandrographolide on front lipocyte proliferation:
Cell survival rate (%)=OD experiment/OD blank × 100%
Experimental result:
Neoandrographolide on 3T3-L1 before the impact of lipocyte proliferation:
Observation Fig. 1 discovery, with the increase of Neoandrographolide concentration, before 3T3-L1, lipocyte proliferation vigor declines, and illustrates that before Neoandrographolide is to 3T3-L1, the propagation of adipocyte is inhibited; Act on respectively after 24h, 48h and 72h the IC of cell half-inhibition concentration 50be respectively 16.4 μ g/mL, 13.0 μ g/mL and 8.6 μ g/mL.
Test two: observe the impact of Neoandrographolide on front Adipocyte Differentiation, one-tenth fat, steatolysis, investigate it and whether can suppress front Adipocyte Differentiation, and promote lipid to decompose.
Experiment material:
Adipocyte before 3T3-L1, purchased from Chinese Academy of Sciences's cell bank;
Neoandrographolide (purity >98%), purchased from National Institute for Food and Drugs Control;
Oil red O, purchased from Aladdin reagent company;
Bradford determination of protein concentration kit is purchased from green skies biotech company, and glycerine is measured kit, Triglyceride Reagent box purchased from Beijing Puli's lema gene technology Co., Ltd.
Experimental technique:
Front Adipocyte Differentiation method: use trypsinization cell monolayer, be made into individual cells suspension with fresh culture, with 4 × 10 4the density in individual/hole is inoculated in 12 orifice plates.100% converges two days later, break up and culture medium was changed to induction culture medium (containing 10% hyclone, 1% dual anti-, 10 μ g/mL insulin, 1 μ mol/L dexamethasone, 0.5mmol/L IBMX in DMEM high glucose medium) in the 0th day, after 48h, break up and within the 2nd day, change secondary induction culture medium (containing 10% hyclone, 1% dual anti-, 10 μ g/mL insulin in DMEM high glucose medium) into, subsequently, every 48h changes basal medium one time.
Oil red O stain method: Cell Differentiation 0th～3d adds gradient concentration Neoandrographolide in culture medium, 8d adopts oil red O stain method observation of cell fat to drip accumulation.Concrete grammar is as follows: the old culture medium of sucking-off, and every hole is washed 1 time with PBS, adds l mL4% formaldehyde to fix 15min, ultrapure washing 3 times; Every hole adds 500 μ L oil red O stain liquid, puts room temperature dyeing 30min on shaking table, and ultrapure washing 3 times, observes Taking Pictures recording under inverted microscope; Every hole adds 500 μ L isopropyl alcohols, puts room temperature extracting 15min on shaking table; Extract 200 μ L/ holes are placed in to 96 orifice plates, and 510nm measures light absorption value, calculates Cell Differentiation rate, and computing formula is as follows:
Cell Differentiation rate %=OD processed group/ OD control group× 100%
The mensuration of content of triglyceride: select the cell of exponential phase, by individual cells suspension, with 3 × 10 5the density in individual/hole is inoculated in 6 orifice plates, according to preceding method Cell differentiation inducing activity.Cell Differentiation 0-3d adds gradient concentration andrographolide in culture medium, when 6d, inhales and abandons culture medium, washes after 3 times with PBS, adds 200 μ L cell pyrolysis liquids, by triglycerides and protein content in kit method mensuration cell.Content of triglyceride in cell, computing formula is as follows:
Figure BDA0000454509570000081
The mensuration that lipid decomposes: after cell induction differentiation, when 6d, be larger fat drop in cell, can be used for measuring lipolysis.Culture medium is abandoned in suction, washes 3 times with PBS buffer solution.The basal medium that contains gradient concentration Neoandrographolide is added to every hole, cultivate 48h.Culture medium is abandoned in suction, with PBS flushing 3 times, then adds containing Hank ' the s buffer solution of 1% bovine serum albumin(BSA) and hatches 3h, by protein content in triglycerides and cell in kit method mensuration buffer solution.Glycerol content in culture medium, computing formula is as follows:
Figure BDA0000454509570000082
The mensuration of protein concentration: adopt Bradford determination of protein concentration kit measurement, reference material is 5mg/mL bovine serum albumin(BSA).
Experimental result:
The impact of Neoandrographolide on front Adipocyte Differentiation, one-tenth fat and steatolysis:
Fig. 2 shows that Neoandrographolide can effectively suppress Adipocyte Differentiation, and in figure, AG.2.5 represents that Neoandrographolide content is 2.5 μ g/mL; AG.5 represents that Neoandrographolide content is 5 μ g/mL; AG.10 represents that Neoandrographolide content is 10 μ g/mL.The Neoandrographolide of 10 μ g/mL dosage, reaches 18.5% to the inhibiting rate of Adipocyte Differentiation, and effect is remarkable.
Fig. 3 shows, by the cell of differentiation is carried out to vitality test, find that low concentration Neoandrographolide is lower on the cell viability impact of differentiation, with MDI processed group there was no significant difference, shows that low dosage Neoandrographolide (being less than 20 μ g/mL) is safe to adipocyte.In figure, control is control group, and AG.20 represents that Neoandrographolide content is 20 μ g/mL; AG.40 represents that Neoandrographolide content is 40 μ g/mL.
Fig. 4 shows that andrographolide can effectively reduce the content of triglycerides in cell, and is concentration dependence, and 10 μ g/mL(are AG.10) inhibiting rate of dosage reaches 33%.The enhancing of lipolysis can reduce gathering of fat, and Neoandrographolide shows the effect of very strong promotion adipocyte steatolysis.
Fig. 5 shows that in 10 μ g/mL dosage processed group culture mediums, glycerol content is 1.45 times that contrast.Above result shows, Neoandrographolide can effectively suppress Adipocyte Differentiation and become fat, promotes lipid to decompose, rather than because due to damaging cells.
Test three: observe the impact of Neoandrographolide on obesity mice body weight, investigate the weight that whether can reduce obesity mice liver and adipose tissue.
Experiment material:
Clean level C57BL/6 mouse, purchased from Shanghai Slac Experimental Animal Co., Ltd.;
Neoandrographolide (purity >98%), purchased from National Institute for Food and Drugs Control.
Experimental technique:
Modeling and intervention: before experiment, 65 C57BL/6 mouse are given under experimental enviroment to the conventional raising of basal feed 5 days, after conforming, be divided at random 12 of low fat control groups, 53 of hyperlipidemia model groups.Low fat control group gives 10% low fat feed (LFD), and all the other hyperlipidemia model groups all give 45% high lipid food (HFD).After 2 weeks, eliminate in hyperlipidemia model group insensitive 5 mouse of hyperlipidemia model according to body weight, 48 remaining mouse are divided into 4 groups, 12 every group at random.Mice group, administration and feed arrange the test grouping in Table 1:C57BL/6 mouse prevention of obesity model.
Table 1
Figure BDA0000454509570000091
Figure BDA0000454509570000101
note: 2% Tween 80 hydrotropy for Neoandrographolide.
Feeding environment and operation: 4, the every cage of mouse; Environment temperature (22 ± 2) ℃; Humidity 40-70%; Natural lighting; Mouse ad lib and drinking-water; Record body weight and food ration every day, the intervention experiment of Neoandrographolide continues 10 weeks.
Experiment finishes, and respectively organizes mouse fasting 12h.Weigh, pluck eyeball blood sampling, put to death, get liver, adipose tissue, and weigh.Body weight gain rate, food utilization computing formula are as follows:
Figure BDA0000454509570000102
Experimental result:
The impact of Neoandrographolide on Mouse Weight, food ration:
Fig. 6 demonstration, the increased weight of 3 concentration administration groups of Neoandrographolide is all starkly lower than hyperlipidemia model group, and presents certain dosage effect.Neoandrographolide 150mg/kg(AG.H) is the most remarkable to the inhibition of high fat diet inducing obesity, and hyperlipidemia model group body weight increment rate is 26.7 ± 3.1%, and AG.H group body weight increment rate is only 8.7 ± 1.1%.
Fig. 7 shows that food ration (Food intake), the food utilization (Food utilization rate) of Neoandrographolide to each experimental group C57 mouse all has certain influence, and the food ration of 3 dosage groups all decreases compared with hyperlipidemia model group.Neoandrographolide is remarkable to the impact effect of food utilization, the food utilization of low dose group be high fat group 75.9%, middle dosage group is 53.7%, high dose group is only 31.8%, showing the main intervention effect of Neoandrographolide to Mouse Weight, is not by suppressing food ration.
The impact of Neoandrographolide on mouse liver, adipose tissue weight:
Fig. 8 (A) shows, high lipid diet has remarkable impact to mouse liver, and hyperlipidemia model group is compared with low fat model group mouse, and liver weight significantly increases, low dosage Neoandrographolide has certain influence to liver weight, and middle and high dosage can effectively reduce the hepatomegaly of meals.Fig. 8 (B) shows that AG.H group is compared with HFD group, and fatty gross weight declines 51.1%, and effect is remarkable.
Test four: observe the impact of Neoandrographolide on obesity mice blood sugar, triglyceride (TG) and T-CHOL (TC), investigate the serum free fatty acid levels that whether can improve obesity mice.
Experiment material:
Clean level C57BL/6 mouse, purchased from Shanghai Slac Experimental Animal Co., Ltd.;
Neoandrographolide (purity >98%), purchased from National Institute for Food and Drugs Control.
Free fatty (FFA) is measured kit, builds up bio tech ltd purchased from Nanjing.
Experimental technique:
The modeling of C57BL/6 mouse and interference method are with test three.
The mensuration of C57BL/6 mice serum index: the centrifugal 15min of 3000r/min after mouse blood room temperature leaves standstill, separation of serum, packing ,-20 ℃ are frozen.Utilize automatic clinical chemistry analyzer to measure blood sugar, triglycerides (TG), T-CHOL (TC).Measure serum FFA according to detection kit.
Experimental result:
The impact of Neoandrographolide on mouse blood sugar, blood fat:
As shown in Fig. 9 (A), high lipid diet can significantly improve 43.0% mouse blood sugar level, the action effect that the mouse blood sugar level after low dosage Neoandrographolide of giving obviously reduces by 31.8%, 3 dosage group is suitable, shows that the Neoandrographolide of low dosage can effectively be controlled mouse blood sugar.As Fig. 9 (B) shows, high lipid diet significantly improves 15.9% mice serum cholesterol levels, and high dose Neoandrographolide can effectively reduce by 12.9% serum cholesterol level, in, low dosage effect is not remarkable.Fig. 9 (C) demonstration, high lipid diet significantly improves 74.3% mice serum triglyceride levels, and AG.M group and AG.H group reduce respectively 17.8%, 31% compared with HFD group, and low dosage effect is not remarkable.Fig. 9 (D) shows, high lipid diet does not make significant difference to mice serum free fatty acid levels, give mouse free fatty acid levels after Neoandrographolide and raise, AG.M group, AG.H group compared with HFD component you can well imagine high 2.6 times, 3.0 times.
Test five: observe the impact of Neoandrographolide on mouse adipose tissue, liver cell form, investigate it and whether can suppress adipose tissues growth, suppress cytolipin accumulation, promote adipocyte smaller volume.
Experiment material:
Clean level C57BL/6 mouse, purchased from Shanghai Slac Experimental Animal Co., Ltd.;
Neoandrographolide (purity >98%), purchased from National Institute for Food and Drugs Control;
Oil red O, purchased from Aladdin reagent company.
Experimental technique:
The modeling of C57BL/6 mouse and interference method are with test three.
C57BL/6 mouse epididymis is the observation by light microscope of adipose tissue around: 1. get epididymis adipose tissue one fritter around, 10% formalin is fixed, and makes tissue fixing completely, abundant; 2. the hepatic tissue blocking fixing is dewatered, transparent; 3. waxdip, embedding section; 4. haematoxylin dyeing; 5. neutral gum mounting drying; 6. under 200 power microscopes, observe the adipocyte state of adipose tissue.
The observation by light microscope of C57BL/6 mouse liver: 1. get every group of same leaf liver one fritter of mouse, be cut into the thickness of 6～8 μ m with freezing-microtome; 2. the fixing 20min of 4% formaldehyde, oil red O stain is processed 30min; 3. rinse unnecessary dyeing liquor with 60% isopropyl alcohol, distilled water rinsing; 4. glycerin gelatine mounting; 5. under 400 power microscopes, observe liver organization pathology and Fat Accumulation situation.
Experimental result:
The impact of Neoandrographolide on mouse adipose tissue, liver cell form:
As shown in figure 10, LFD group, adipocyte small volume, form is clear, structural integrity; The epididymis of HFD group around adipocyte volume expands, and cell size is inhomogeneous, irregular arrangement; The each dosage group of Neoandrographolide is compared with HFD group, and adipocyte volume obviously reduces, in 200 power microscope field range fat cell hypertrophy number showed increased; AG.H group adipocyte volume is even less than low fat control group, the compact rule of cell arrangement, and form is clear.These results show that Neoandrographolide is remarkable to the inhibition of C57 mouse fat tissue cell lipid accumulation.
Finally, the present invention can summarize with other the concrete form without prejudice to spirit of the present invention and principal character.Therefore, no matter from that, above-mentioned embodiment of the present invention all can only think explanation of the present invention can not limit the present invention, claims have been pointed out scope of the present invention, and scope of the present invention is not pointed out in above-mentioned explanation, therefore, any change in implication and the scope suitable with claims of the present invention, all should think to be included in the scope of claims.

Claims (8)

1. a Neoandrographolide is in the application of preparing in diet food or medicine.
2. the pharmaceutical composition of an acceptable additives composition using Neoandrographolide as active component and in pharmaceutics is in the application of preparing in diet food or medicine.
3. application as claimed in claim 2, is characterized in that, described pharmaceutical composition is tablet, is made up of the raw material of following mass percent:
Figure FDA0000454509560000011
4. application as claimed in claim 3, is characterized in that, described filler is one or more in starch, microcrystalline cellulose, lactose.
5. application as claimed in claim 3, is characterized in that, described disintegrant is one or more in sodium carboxymethyl starch, Ac-Di-Sol, PVPP, low-substituted hydroxypropyl cellulose, dried starch.
6. application as claimed in claim 3, is characterized in that, described lubricant is one or more in talcum powder, dolomol, superfine silica gel powder, hydrogenated vegetable oil.
7. the application as described in claim as arbitrary in claim 3～6, is characterized in that, described tablet is made up of the raw material of following mass percent:
Figure FDA0000454509560000012
8. application as claimed in claim 7, is characterized in that, slimming medicine prepared by described pharmaceutical composition, and its consumption is 50～150mg/ days.
CN201410009603.8A 2014-01-08 2014-01-08 Application of andrographolide C to preparation of weight-losing food or medicine Pending CN103766901A (en)

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CN113332275A (en) * 2021-05-20 2021-09-03 中国中医科学院广安门医院 Application of andrographolide in preparing product for promoting browning of white fat
US11318153B2 (en) 2018-06-25 2022-05-03 Bialpha International Sdn. Bhd. Method of using Neoandrographolide for lowering blood sugar, lowering blood lipid, improving liver function and improving renal function
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