CN102133221A - Compound traditional Chinese medicine extract preventing glucose metabolism disturbance and preparation method thereof - Google Patents
Compound traditional Chinese medicine extract preventing glucose metabolism disturbance and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a compound traditional Chinese medicine extract preventing glucose metabolism disturbance, comprising the following effective ingredients: dried alcohol, Beta-sitosterol, hexacosanoic acid, butenolide III, oleanolic acid, berberine, jateorhizine, coptisine, salvianic acid A, salvianolic acid B, ring-tetracosane, 9,12-octadecadienoic acid, 5,7-dimethoxy coumarin, specnuezhenide, ginsenoside Rb1 and Rg1, notoginsenoside R1 and encommiol. A preparation method of the extract is as follows: extracting raw material medicines by C1-3 alcohol and/or water, combining a total extract, extracting the total extract by organic solvents with different polarities so as to obtain all effective ingredients, and finally mixing the ingredients so as to obtain a product. As for the compound traditional Chinese medicine extract, a large number of ineffective chemical ingredients in Chinese medicine are removed, so that the content of effective ingredients is increased greatly, and the influence on product processing and preparation quality caused by the ineffective ingredients is reduced; and simultaneously, the preparation process is stable, the product quality is controllable, the mass production is facilitated, and the drug effect of the compound traditional Chinese medicine is improved.
Description
Technical field
The present invention relates to the herbal mixture field, be specifically related to a kind of herbal mixture extract of preventing and treating carbohydrate metabolism disturbance and preparation method thereof.
Background technology
Diabetes are the metabolic diseases that cause because of the blood glucose metabolism disorder.Diabetes have become the significant threat of contemporary society's human health, have caused the great attention of WHO and national governments, are the focuses of medical scientific research.But so far the pathogenesis of blood glucose metabolism disorder is not also illustrated, the clinical treatment diabetes also do not have ideal way.Though chemical drugs has many medicines to blood sugar lowering, its comprehensive therapeutic effect is unsatisfactory, and the expense that chemicals also has untoward reaction and medication also is the drawback of chemical drugs than problems such as height.
Clinical practice shows, Chinese medicine is preventing and treating aspect the diabetes, because its clinical integral viewpoint and dialectical opinion are administered opinion, and Chinese medicine has multicomponent, can many target spots, multi-level performance drug effect, so clinical efficacy is definite, have advantage safely and effectively.
Chinese patent 200410051250.4 discloses a kind of medicine for the treatment of hyperlipemia, and called after " the loyal art of compound recipe is transferred fat side " (FTZ) is made up of Fructus Ligustri Lucidi, the Rhizoma Atractylodis Macrocephalae, the Cortex Eucommiae, Radix Notoginseng, Radix Cirsii Japonici, Radix Salviae Miltiorrhizae, Rhizoma Coptidis, Fructus Citri Sarcodactylis eight flavor Chinese medicines.Be through application experience side of clinical more than ten years, be used for the treatment of blood glucose metabolism disorder diseases such as diabetes and obtain desirable curative effect that experimentation shows that also this side has good blood sugar reducing function to experimental rat diabetes, this prescription patent is obtained the authorization.
Though this compound recipe has the effect of well-tuned blood glucose, but, form its complex chemical composition by eight flavor Chinese medicines owing to be Chinese medicine compound, effective ingredient is also unclear, its product quality is difficult to obtain effective control in the practical application, finally influences clinical efficacy stable of product, and invalid component is too many when being used for useful in preparing drug formulations, dose is big, dosage form can not be diversified, is unfavorable for the large-scale production listing, and extensive use is restricted.
Summary of the invention
The objective of the invention is to that its product quality is difficult to obtain effective control in, the practical application unclear according to existing complex chemical composition, the effective ingredient that is used for preventing or treats the herbal mixture of blood glucose metabolism disorder to exist, defectives such as invalid component is too many provide a kind of herbal mixture extract of preventing and treating carbohydrate metabolism disturbance.
Another object of the present invention provides above-mentioned herbal mixture preparation method of extract.
The object of the invention is achieved by the following technical programs:
A kind of herbal mixture extract (FTZCE) of preventing and treating carbohydrate metabolism disturbance, effective ingredient is ceryl alcohol, cupreol, n-hexacosanoic acid, atractylenolide, oleanolic acid, berberine, jateorhizine, coptisine, danshensu, salvianolic acid B, cyclolignocerane, 9,12-octadecadienoic acid, 5,7-dimethoxy coumarin, specnuezhenside, ginsenoside Rb1 and Rg1, arasaponin R1, eucommoil.
The part by weight of above-mentioned herbal mixture extract effective ingredient is 1～5:1～6:1～4:1～4:1～8:1～8:1～5:1～5:1～6:1～5:1～4:1～3:1～5:1～4:1～8:1～8:1～3:1～4.
Above-mentioned herbal mixture preparation method of extract comprises the steps:
(1) crude drug Radix Salviae Miltiorrhizae, Fructus Ligustri Lucidi, Rhizoma Coptidis, Radix Cirsii Japonici, the Cortex Eucommiae, the Rhizoma Atractylodis Macrocephalae, Radix Notoginseng and Fructus Citri Sarcodactylis are carried out C
1-3Alcohol extraction and/or water are carried, and merge total extract;
(2) total extract that obtains with the organic solvent extraction step (1) of opposed polarity obtains the effective site of opposed polarity, and each effective site is mixed, and obtains preventing and treating the herbal mixture extract of carbohydrate metabolism disturbance.
C
1-3Alcohol is methanol, ethanol or propanol.
The preferred concrete steps of above-mentioned preparation method are:
(1) crude drug Radix Notoginseng and Fructus Ligustri Lucidi are carried out C
1-3Alcohol extraction obtains C
1-3Ethanol extract carries out water with Radix Cirsii Japonici, the Rhizoma Atractylodis Macrocephalae, Radix Salviae Miltiorrhizae, the Cortex Eucommiae, Fructus Citri Sarcodactylis and Rhizoma Coptidis and carries, concentrates, and obtains water extract, merges C
1-3Ethanol extract and water extract obtain total extract;
(2) total extract that obtains with petroleum ether extraction step (1) obtains effective extract part A;
(3) then with extract remaining behind ethyl acetate extraction step (2) petroleum ether extraction, obtain effective extract part B;
(4) and then with extract remaining behind n-butanol extraction step (3) ethyl acetate extraction, obtain effective extract part C, merge each effective site, obtain preventing and treating the herbal mixture extract of carbohydrate metabolism disturbance.
As a kind of preferred version, described C
1-3The ethanol extraction of alcohol drawings 30～95 volume % 1～5 time, preferred 2～4 times, each ethanol volume that extracts is more than 1.5 times of quality of medicinal material, preferred 5～12 times, each extraction time is 5 min～5 h, preferred 1～3 h; Described water is carried preferably and being boiled 1～5 time with decocting, and preferred 2～4 times, the volume of each water is more than 1.5 times of quality of medicinal material, and preferred 6～12 times, each decocting time is 5 min～5 h, preferred 0.5～4 h; Concentrated solution volume after described the concentrating is 0.2～5 times of quality of medicinal material, preferred 1～2 times.
In the above-mentioned preparation method, effectively the preparation method of extract part A is as follows: add 1～15 times, preferred 1～5 times to the petroleum ether of extract volume, extract 1～5 time, preferred 2～4 times, combining extraction liquid obtains position A through cold drying; Effectively the preparation method of extract part B is as follows: add 1～15 times, preferred 3～10 times to the ethyl acetate of extract volume, extract 1～5 time, preferred 2～4 times, combining extraction liquid obtains position B through cold drying; Effectively the preparation method of extract part C is as follows: add 1～15 times, preferred 3～10 times to the n-butyl alcohol of extract volume, extract 1～5 time, preferred 2～4 times, combining extraction liquid obtains position C through cold drying.
In the herbal mixture that the present invention prepares, the weight ratio of effective site A, B and C is 0～1:0～2:0～3.
Among the present invention, can also be processed into forms such as oral formulations or ejection preparation with each effective site or its compositions according to acceptable carrier on the pharmaceutics and general formulation method.
Compared with prior art, the present invention has following beneficial effect:
By preparation method of the present invention, can remove a large amount of invalid chemical constituents in the Chinese medicine, obtain effective extract part of opposed polarity, both kept the Chinese medicine characteristic, the active constituent content in the Chinese medicine is improved greatly, reduced the influence of invalid components the product processing and the quality of the pharmaceutical preparations, make this herbal mixture effective ingredient clear and definite, stable preparation process, controllable product quality helps suitability for industrialized production.Experimentation shows that the pharmaceutical composition that the inventive method obtains has good blood sugar reducing function to experimental rat diabetes.
The specific embodiment
Further explain the present invention below in conjunction with embodiment, but embodiment does not do any type of qualification to the present invention.
The preparation of the total extract (FTZ) of embodiment 1 Chinese medicine compound
Radix Notoginseng, Fructus Ligustri Lucidi two medicated powder in Fructus Ligustri Lucidi, the Rhizoma Atractylodis Macrocephalae, the Cortex Eucommiae, Radix Notoginseng, Radix Cirsii Japonici, Radix Salviae Miltiorrhizae, Rhizoma Coptidis, the Fructus Citri Sarcodactylis (weight ratio is 1:1:1:2:1:2:1:1) are broken into coarse powder, distinguish reflux, extract, three times with 12 times, 10 times of medical material amount and 8 times 60 volume % ethanol, each 2 h, merge extractive liquid,, reclaiming ethanol and making alcohol extraction part concentration is that 1 ml is equivalent to 1 g crude drug; All the other medical materials decoct respectively and extract three times all with the water of 12 times, 10 times and 8 times amounts in the side, and each 2 h merge three times the water extract, and vacuum decompression is concentrated into 1 g medical material/ml extracting solution, and water is carried partly and the front alcohol extraction partly merges and obtains the Chinese medicine compound total extract.
The preparation of the extract part of embodiment 2 compound recipe total extracts
With the petroleum ether extraction of 5 times of volumes of compound recipe total extract once, separate petroleum ether layer; Extract twice with 3 times of volumes again, separate obtaining petroleum ether layer, merge three times petroleum ether extraction liquid, reclaim petroleum ether, again in 0.08 MPa, 65 ℃ of vacuum dryings obtain effective extract part A;
The remaining extract in extraction back adds the ethyl acetate extraction of 10,8,5 times of amounts more respectively, and solvent is reclaimed at the ethyl acetate extraction position that obtains, and in 0.07 MPa, 75 ℃ of following vacuum dryings obtain effective extract part B;
Extract remaining behind the ethyl acetate extraction extracts respectively with the n-butyl alcohol of 10,6,3 times of amounts of total extract respectively again, obtains extract and merges, and reclaims n-butyl alcohol, and in 0.07 MPa, 75 ℃ of following vacuum dryings obtain compound recipe extract part C with extract.
According to total inventory of Chinese medicine compound and the amount that obtains each extract part, calculate the yield of each extract respectively.
Effectively the yield of extract part A is 0.10%～0.30%
Effectively the yield of extract part B is 0.40%～1.00%
Effectively the yield of extract part C is 2.00%～4.00%.
The herbal mixture extract is that 0～1:0～2:0～3 mix by the weight ratio of effective extract part A, B and C.Contain ceryl alcohol, cupreol, n-hexacosanoic acid, atractylenolide, oleanolic acid, berberine, jateorhizine, coptisine, danshensu, salvianolic acid B, cyclolignocerane, 9 after measured, 12-octadecadienoic acid, 5,7-dimethoxy coumarin, specnuezhenside, ginsenoside Rb1 and Rg1, arasaponin R1, eucommoil etc., the part by weight of each component is 1～5:1～6:1～4:1～4:1～8:1～8:1～5:1～5:1～6:1～5:1～4:1～3:1～5:1～4:1～8:1～8:1～3:1～4.
The capsular preparation method of embodiment 3 extractive compositions
Extract makes up according to following weight portion:
Effectively extract part A is 1 part
Effectively extract part B is 2 parts
Effectively extract part C is 5 parts
With each extract of aforementioned proportion, effectively the starch mix homogeneously of extract part A and equivalent mixes with effective extract part B again, mixes with effective extract part C at last, dry granulation behind three's mix homogeneously, behind the drying under reduced pressure, granulate, fill No. 2 capsule, polishing, quality inspection packing.
Measure the capsule that makes with the HPLC method, it contains each active constituent content and the results are shown in Table 1:
Table 1 FTZCE capsule HPLC assay result
Embodiment 4 Chinese medicine compound extracts and compositions thereof are to the effect of normal mouse blood sugar
Animal: 60 of healthy NIH mices, male and female half and half, body weight are 18～22 g.
Grouping: be divided into 8 groups at random: blank group, glibenclamide group, 1 group, 2 groups, 3 groups of compound recipe total extract group (being called for short FTZ), extract part compositionss (weight ratio of A, B and C is 1:2:3, is called for short FTZCE), 10 every group by sex.
Experimental technique: behind animal fasting 12 h, get blood from the eye socket venous plexus, the centrifugal 10min of blood sample 3000 rpm, separation of serum by the operation of glucose kit description, is measured blood sugar content.Get and irritate stomach immediately behind the blood and give to supply accordingly the reagent thing, glibenclamide 50 mg/kg wherein, the blank group gavages the co-content normal saline, and each administration volume is 0.2 mL/10g body weight.After administration when 3 h, 6 h and 9 h, measuring blood sugar of blood extracting content as stated above respectively, experimental result sees Table 2.Result's credit by statistics analyses and selects the t method of inspection for use, relatively the difference of each administration group and matched group.
Table 2 FTZCE is to the effect of normal mouse blood sugar (n=10, x ± s)
Annotate: with matched group ratio, * P＜0.05, * * P＜0.01.
Experimental result shows, with the normal mouse ratio, in administration each group of positive control drug, total extract and compound extract compositions after 3 hours; After 3,6,9 hours, positive control drug, total extract, extract part have significant reduction effect to the blood glucose of normal mouse.Each organize administration after 6 hours hypoglycemic effect the most remarkable.
Embodiment 5 FTZCE are to the influence of the blood glucose of diabetic mice
Medicine and reagent: FTZCE(herbal mixture extractive composition, the weight ratio of A, B and C is 1:2:3), blood glucose test kit (the safe reagent company limited of Beijing northization), streptozotocin (STZ, U.S. Sigma company product), quench one's thirst the peace capsule (Tonghua Baishan Pharmaceutical Ltd's product).
Adopt Advantage electronic induction blood glucose meter and supporting paper slip (Switzerland Roche company product) to measure blood glucose.
Experimental subject is to economize the cleaning level NIH mice that Experimental Animal Center provides, male and female half and half, body weight 18～22 g by Nanfang Medical Univ.
(a) to the influence of the blood glucose of streptozotocin (STZ) type diabetic mice
With 80 healthy NIH mices, numbering is divided into 8 groups after weighing at random, 10 every group, gets wherein 1 group as the normal control group.All the other respectively organized the mice fasting after 16 hours, the freshly prepared 1% streptozotocin solution of lumbar injection 150 mg/kg, after 72 hours, the ophthalmic corner of the eyes is got blood, adopt Advantage electronic induction blood glucose meter and supporting paper slip (Switzerland Roche company product) to measure blood glucose, blood glucose value is defined as the diabetes model Mus greater than 16.7 mmol/L persons.The diabetes model Mus is divided into 7 groups at random by after the blood glucose value numbering, 3 treatment groups of FTZCE (30,60,120 mg/kg), 2 groups of compound recipe total extract (FTZ) (250,500 mg/kg), the peace of quenching one's thirst capsule for treating group (500 mg/kg), model control group.Each group difference gastric infusion 1 time/day, successive administration 30 days was got blood from mice socket of the eye venous plexus respectively in 2 hours, centrifugal 10 min of blood sample 3000 rpm after the last administration played animal fasting, administration in preceding 12 hours, separation of serum is measured blood sugar content by the glucose kit description.Experimental result sees Table 3.
Table 3. FTZCE is to the influence of the blood glucose of streptozotocin (STZ) type diabetic mice (n=10, x ± s)
Annotate: compare with normal group,
*P＜0.05,
*P＜0.01; Compare with model group,
△P＜0.05,
△ △P＜0.01; With comparing before the treatment
▲P＜0.05,
▲ ▲P＜0.01.
(b) influence of the blood glucose of the hyperglycemia mice that epinephrine is caused
With 60 healthy NIH mices, after weighing, numbering is divided into 6 groups at random, every group 10, get wherein 1 group as normal control group, epinephrine group, the little heavy dose of FTZCE (30,60mg/kg) group, FTZ 500 mg/kg group, the peace of quenching one's thirst capsule for treating group (500 mg/kg).Each group difference gastric infusion 1 time/day, normal control group and epinephrine group give the equal-volume normal saline, successive administration 7 days, behind last administration 2 h, matched group lumbar injection (ip) equal-volume normal saline, all the other respectively organize ip epinephrine (240 mg/kg) solution, and 0.5 h, 1 h get blood from mice socket of the eye venous plexus behind the ip epinephrine respectively, separation of serum, mensuration blood glucose.Experimental result sees Table 4.
The influence of the blood glucose of the hyperglycemia mice that table 4. FTZCE causes epinephrine (n=10, x ± s)
(c) influence of the blood glucose of the hyperglycemia mice that glucose is caused
With 60 healthy NIH mices, after weighing, numbering is divided into 6 groups at random, and 10 every group, normal control group, the high sugared model group of glucose, the little heavy dose of FTZCE (30,60 mg/kg) group, FTZ 500 mg/kg group, the peace of quenching one's thirst capsule for treating group (500mg/kg).Each group difference gastric infusion 1 time/day, normal control group and glucose group give the equal-volume normal saline, successive administration 7 days, behind last administration 2h, matched group ip equal-volume normal saline, all the other respectively organize ip glucose (2 g/kg) solution, and 0.5,1,2 h get blood from mice socket of the eye venous plexus behind the ip glucose respectively, separation of serum, mensuration blood glucose.Experimental result sees Table 5.
The influence of the blood glucose of the hyperglycemia mice that table 5. FTZCE causes glucose (n=10, x ± s)
Annotate: compare with normal group,
*P＜0.01 is compared before treating,
▲P＜0.05,
▲ ▲P＜0.01.
Embodiment 6FTZCE brings out the influence of the plain resistance of mouse islets to hydrocortisone
Be divided into 4 groups at random after 40 mices numbering weighed, every group 10, get wherein one group of subcutaneous injection normal saline as normal control group, one group of subcutaneous injection hydrocortisone (36 mg/kg), two groups of subcutaneous injection hydrocortisone (36 mg/kg) are irritated the little heavy dose of stomach FTZCE (30,60 mg/kg) more respectively in addition.Each group difference gastric infusion 1 time/day, normal control group and hydrocortisone group give the equal-volume normal saline, successive administration 10 days, behind last administration 2 h, matched group ip equal-volume normal saline, all the other respectively organize ip insulin (0.5 g/kg), and 0.5 h, 2 h pluck eye and get blood behind the ip glucose respectively, and separation of serum, automatic clinical chemistry analyzer are measured blood glucose.
The result is shown in table 6, table 7.
Table 6 FTZCE brings out influence (n=10, the x ± s) of the plain resistance of mouse islets to hydrocortisone
Table 7 FTZCE brings out influence (n=10, the x ± s) of the plain resistance of mouse islets to hydrocortisone
Annotate: compare with normal group,
*P＜0.05,
*P＜0.01; Before treating, compare,
▲P＜0.05,
▲ ▲P＜0.01.
Embodiment 7FTZCE is to the effect of streptozotocin type diabetes model rat
Medicine and reagent: streptozotocin (STZ), U.S. Sigma company product; Sodium citrate, Shantou City's brilliance laboratory product, lot number 20000116.
Experimental subject is 36 of female Wistar regular grade rats, body weight 220 ~ 250 g.(No.1 Military Medical Univ.'s Experimental Animal Center provides, the quality certification number: Guangdong probatio inspectionem pecuoarem word 2004B023 number)
After all rat adaptabilities are fed a week, fasting 12h, getting 7 rats is normal control group (abbreviation matched group), other rat disposable celiacs are injected 50 mg/kg STZ, and (STZ faces the sodium citrate buffer preparation with preceding usefulness 0.1 mol/L, pH4.5, use up in 10 min), matched group is then injected the equivalent liquor sodii citratis.Blood glucose FBG is a hyperglycemia diabetes model rat greater than 16.7 mmol/L persons behind the 72h.Diabetes rat is divided into diabetic model group, the big small dose group of FTZCE at random.Big agent group by people and rat dose,equivalent than mg/5 mL/ kg administration every days 30, little dose of group mg/5 mL/kg every days 10, model group and normal group are given 5 mL/kg distilled water every day, irritate stomach the morning once, treat 15 days.Before each organizes modeling, after the modeling, the docking of treatment back gets blood and surveys blood glucose (FBG), serum insulin (Ins), get the rat pancreas at last and carry out HE dyeing and immunohistochemical staining as specimen.
Adopt Advantage electronic induction blood glucose meter and supporting paper slip (Switzerland Roche company product) to measure blood glucose.The blood glucose glucose oxidase method, serum insulin is with putting the method for exempting from.
(x ± s) expression carries out significance test with variance analysis method to experimental data with means standard deviation.
Influence to blood glucose the results are shown in Table 8, and model group and normal group compare, and FBG significantly raises (P＜0.01), significantly descends (P＜0.01) with model group comparison FBG after the big or small dosage treatment, illustrates that FTZCE has remarkable hypoglycemic activity.
Table 8 FTZCE respectively organizes rat blood sugar change list (mmol/L) to the STZ model
Annotate: compare with normal group,
*P＜0.05,
*P＜0.01; Compare with model group,
△P＜0.05,
△ △P＜0.01; With comparing before the treatment
▲ ▲P＜0.01.
Influence to serum insulin the results are shown in Table 9, and model group and normal group compare, and Ins significantly reduces (P＜0.01), and serum Ins level shows that than the remarkable rising of model group (P＜0.05) secretion has certain facilitation to FTZCE to Ins after the big or small dosage treatment.
Table 9 FTZCE respectively organizes rat blood serum insulin change list (mU/L) to the STZ model
Annotate: compare with normal group,
*P＜0.05,
*P＜0.01.Compare with model group
▲ ▲P＜0.01.
HE dyeing histological observation is found: the normal group islets of langerhans is circular, elliptical erythrocyte group, and boundary is clear, and islets of langerhans number and island inner cell number are more, and kytoplasm is abundant; STZ model group islets of langerhans number and island inner cell number reduce, and cellular morphology is irregular, and nuclear differs in size, form differs, the part karyopycnosis, and a few cell is the cavity shape; Heavy dose of group islets of langerhans number and island inner cell are counted showed increased, and cell distribution is even, and the nuclear size is equal substantially, no karyopycnosis; Little dose of group islets of langerhans number and island inner cell number increase, and form is rule comparatively, the small part karyopycnosis.This explanation FTZCE has significant protective effect to the islet cells tissue of STZ MODEL DAMAGE, and can promote secretion of insulin.
Embodiment 8Influence to B cell
Cultivate rat Langerhans islet B cell routinely, treat to add 200 μ M H respectively behind the cell fusion
2O
2Educate 1h altogether, the FTZCE (weight ratio of A, B and C is that 1:2:3 is prepared from) and the FTZ that add various variable concentrations then, continue to cultivate 24h after the time, collecting cell is made cell homogenates, with oxide dismutase (SOD), catalase (CAT) and glutathion peroxidase (GSH-Px) and glutathion (GSH) detection kit, adopt colorimetry to detect SOD, CAT and GSH-Px activity and GSH content, study above-mentioned variable concentrations FTZCE B cell SOD, CAT and GSH-Px activity and the active influence of GSH content.
Influence to Bcl-2, PPAR-α and Bax and Fas gene expression in the B cell
Handle B cell with method in addition, extract total RNA in the B cell with Trizol reagent, the RT-PCR method is measured Bcl-2, PPAR-α and Bax and Fas expression conditions in the B cell, observes the influence of FTZCE to Bcl-2, PPAR-a and Bax in the B cell and Fas gene expression.
Statistical method: data are represented with means standard deviation, carry out significance test with variance analysis method.
Experimental result
Influence to B cell SOD, CAT and GSH-Px activity and GSH content
2 ~ 10 μ g/ml concentration FTZCE and FTZ handle back B cell SOD, CAT and GSH-Px activity and GSH content significantly increases (P＜0.01), and the enhancing of obvious dose dependent is arranged, and the results are shown in Table 10.Illustrate that herbal mixture extract (FTZCE) has the effect of significantly improving to B cell SOD, CAT and GSH-Px activity and GSH content, helps antioxidant stress injury.The protection B cell.
Table 10 different pharmaceutical is to the influence of oxidative stress damage B cell SOD, CAT and GSH-Px activity and GSH content
Annotate: 1) compare with the normal control group,
☆ ☆P＜0.01; Compare * P＜0.05 with model group, * * P＜0.01;
2) FTZ: the loyal art of compound recipe is transferred fat formula extraction, FTZCE: herbal mixture extract, BBR: berberine hydrochloride.
Influence to Bcl-2, PPAR-α and Bax and Fas gene expression in the B cell
Bcl-2 and Bax and Fas gene expression are expressed normally in the normal group B cell, add 200 μ M H
2O
2Educate altogether that Bcl-2 and PPAR α gene expression obviously reduce behind the 1h, Bax and Fas gene expression then obviously increase; Anti-apoptotic genes expression Bcl-2 and anti-inflammatory factor PPAR-α gene expression significantly increase (P＜0.01) in the B cell of 2 ~ 10 μ g/ml concentration FTZCE processing back; and apoptogene Bax and Fas gene expression significantly reduce (P＜0.01); illustrate that composition of plant extracts of the present invention (FTZCE) makes anti-apoptotic genes expression Bcl-2 gene expression that obvious facilitation (P＜0.01) be arranged; and the expression of inhibition apoptosis gene Bax and Fas, thereby the protection B cell is avoided oxidative stress damage apoptosis.The results are shown in Table 11, BBR is a berberine hydrochloride.
Table 11 different pharmaceutical is to the influence of Bcl-2, Bax in the B cell and Fas gene expression
Annotate: compare with the normal control group,
☆ ☆P＜0.01; Compare * P＜0.05 with model group, * * P＜0.01.
Conclusion: FTZEE confirms to have the promotion insulin secretion through pharmacological research, and the content that increases serum insulin has blood sugar lowering, improves the metabolic effect of blood glucose.Its mechanism of action may comprise:
(1) FTZEE can strengthen the active and GSH content of B cell superoxide dismutase (SOD), catalase (CAT) and glutathion peroxidase (GSH-Px), can suppress lipid peroxidation and remove hydroxyl radical free radical, antioxidant stress injury promotes B cell regeneration.
(2) FTZEE can increase the cl-2 gene expression of B cell anti-apoptotic factor B and suppress B cell antiapoptotic factors Bax and Fas gene expression, thereby reduces the generation of apoptosis, stimulates normal B cell excreting insulin.
(3) FTZEE can increase B cell anti-inflammatory factor PPAR-α gene expression, and the antagonism role of cytokines reduces the B cell damage.Increase the content of serum insulin by protection, reparation B cell.
Clinical research is the result show, plant extract FTZCE of the present invention has significant blood sugar lowering, improves the blood glucose metabolism, can be used for treating blood glucose metabolism disorder relevant diseases such as hyperglycemia and diabetes.
Claims (10)
1. herbal mixture extract of preventing and treating carbohydrate metabolism disturbance, effective ingredient is ceryl alcohol, cupreol, n-hexacosanoic acid, atractylenolide, oleanolic acid, berberine, jateorhizine, coptisine, danshensu, salvianolic acid B, cyclolignocerane, 9,12-octadecadienoic acid, 5,7-dimethoxy coumarin, specnuezhenside, ginsenoside Rb1 and Rg1, arasaponin R1, eucommoil.
2. according to the described herbal mixture extract of claim 1, the part by weight that it is characterized in that effective ingredient is 1～5:1～6:1～4:1～4:1～8:1～8:1～5:1～5:1～6:1～5:1～4:1～3:1～5:1～4:1～8:1～8:1～3:1～4.
3. the described herbal mixture preparation method of extract of claim 1 comprises the steps:
(1) crude drug Radix Salviae Miltiorrhizae, Fructus Ligustri Lucidi, Rhizoma Coptidis, Radix Cirsii Japonici, the Cortex Eucommiae, the Rhizoma Atractylodis Macrocephalae, Radix Notoginseng and Fructus Citri Sarcodactylis are carried out C
1-3Alcohol extraction and/or water are carried, and merge total extract;
(2) total extract that obtains with the organic solvent extraction step (1) of opposed polarity obtains the effective site of opposed polarity, and each effective site is mixed, and obtains preventing and treating the herbal mixture extract of carbohydrate metabolism disturbance.
4. preparation method according to claim 3 may further comprise the steps:
(1) crude drug Radix Notoginseng and Fructus Ligustri Lucidi are carried out C
1-3Alcohol extraction obtains C
1-3Ethanol extract carries out water with Radix Cirsii Japonici, the Rhizoma Atractylodis Macrocephalae, Radix Salviae Miltiorrhizae, the Cortex Eucommiae, Fructus Citri Sarcodactylis and Rhizoma Coptidis and carries, concentrates, and obtains water extract, merges C
1-3Ethanol extract and water extract obtain total extract;
(2) total extract that obtains with petroleum ether extraction step (1) obtains effective extract part A;
(3) with extract remaining behind ethyl acetate extraction step (2) petroleum ether extraction, obtain effective extract part B;
(4) again with extract remaining behind n-butanol extraction step (3) ethyl acetate extraction, obtain effective extract part C, merge each effective site, obtain preventing and treating the herbal mixture extract of carbohydrate metabolism disturbance.
5. preparation method according to claim 4 is characterized in that described C
1-3Alcohol extraction is meant the C with 30～95 volume %
1-3Alcohol extraction 1～5 time, each C that extracts
1-3The alcohol volume is more than 1.5 times of quality of medicinal material, and each extraction time is 5 min～5 h.
6. preparation method according to claim 4 is characterized in that described water is carried to be meant with decocting and to boil 1～5 time that the volume of each water is more than 1.5 times of quality of medicinal material that each decocting time is 5 min～5 h; Concentrated solution volume after described the concentrating is 0.2～5 times of quality of medicinal material.
7. preparation method according to claim 4, the preparation method that it is characterized in that described effective extract part A is to add 1 ~ 15 times to the petroleum ether of extract volume, extracts combining extraction liquid 1～5 time, reclaim petroleum ether, obtain effective extract part A through cold drying.
8. preparation method according to claim 4, the preparation method that it is characterized in that effective extract part B is to add 1～15 times to the ethyl acetate of extract volume, extracts combining extraction liquid 1～5 time, reclaim ethyl acetate, obtain effective extract part B through cold drying.
9. preparation method according to claim 4, the preparation method that it is characterized in that effective extract part C is to add 1～15 times to the n-butyl alcohol of extract volume, extracts combining extraction liquid 1～5 time, reclaim n-butyl alcohol, obtain effective extract part C through cold drying.
10. preparation method according to claim 4 is characterized in that effective extract part A, B and C and weight ratio are 0～1:0～2:0～3.
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| WO2012094834A1 (en) * | 2011-01-14 | 2012-07-19 | 广东药学院 | Compound traditional chinese medicine extract for prevention and treatment of glycometabolism disorders and preparation method thereof |
| CN106466325A (en) * | 2016-09-30 | 2017-03-01 | 郑州大学第附属医院 | The medicine of a kind of prevention or treatment diabetes, compositionss and its preparation |
| CN116898912A (en) * | 2023-07-12 | 2023-10-20 | 广东药科大学 | Application of compound traditional Chinese medicine in preparation of medicine for preventing and treating diabetic encephalopathy |
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