CN102091083A - Petroleum ether extract of traditional Chinese medicine for preventing and treating glucose and lipid metabolic disturbance and preparation method thereof - Google Patents

Abstract

The invention discloses a petroleum ether extract of traditional Chinese medicine for preventing and treating glucose and lipid metabolic disturbance, comprising the following active components: isoceryl alcohol, beta-sitosterol, n-hexacosane acid, butenolide III, oleanolic acid, berberine, jateorhizine, salvianolic acid B, lignocerane, 9,12-octadecadienoic acid, 5,7-dimethoxy coumarin and ginsenoside Rb1. The preparation method comprises the following steps: after extracting crude medicines of root of red rooted salvia, glossy privet fruit, coptis, thistle, eucommia, white atractylodes rhizome, pseudo-ginseng and finger citron by C1-3 alcohols and/or water, combining all the extracts, and extracting the total extract by petroleum ether to obtain the petroleum ether extract of traditional Chinese medicine for preventing and treating glucose and lipid metabolic disturbance. A big amount of inactive components in the original compound traditional Chinese medicine are removed from the extract provided by the invention, so that much more efficient extraction parts of the traditional Chinese medicines are obtained, and the extract has a stable preparation process, controllable quality and notable curative effect, and is convenient to use and be produced into various dosage forms.

Description

A kind of Chinese medicine ligroin extraction of preventing and treating the glycolipid metabolism disorder and preparation method thereof

Technical field

The present invention relates to the field of Chinese medicines, be specifically related to a kind of Chinese medicine ligroin extraction that is used to prevent and treat the disorderly relevant disease of glycolipid metabolism relevant diseases such as (hyperlipemia, diabetes, metabolic syndrome) animal are atherosis and preparation method thereof.

Background technology

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), contains Fructus Ligustri Lucidi, Radix Cirsii Japonici, the Rhizoma Atractylodis Macrocephalae, Radix Salviae Miltiorrhizae, the Cortex Eucommiae, Fructus Citri Sarcodactylis, can also add Radix Notoginseng and Rhizoma Coptidis.The preferred weight percent of each component is Fructus Ligustri Lucidi 20～25%, Radix Cirsii Japonici 12～15%, the Rhizoma Atractylodis Macrocephalae 10～15%, Radix Salviae Miltiorrhizae 10～15%, the Cortex Eucommiae 10～15%, Fructus Citri Sarcodactylis 10～12%, Radix Notoginseng 10～12%, Rhizoma Coptidis 6～10%.This side is used for the treatment of hyperlipemia, obtains desirable curative effect, and clinical total effective rate reaches 91%.

Though the loyal art of compound recipe transfers fat side that good blood lipid regulation and arteriosclerosis function and clinical efficacy are arranged, but owing to be compound Chinese medicinal preparation, also have many weak points: (1) its composition is complicated, its effective ingredient is clear not enough, be difficult for quantizing, its product quality is difficult for obtaining effectively control in the practical application, influences the stability of product clinical efficacy the most at last; (2) preparation technology is simple relatively, and effective ingredient can not get due purification in the side, and impurity component is many, taking dose big (needing every day more than nearly 5 grams), and it is undesirable that patient takes compliance; (3) because prescription extract amount is big, and various controlled release forms are made in inconvenience, useful in preparing drug formulations can not be diversified, and range of application is subjected to various restrictions.

Summary of the invention

The objective of the invention is to overcome the above-mentioned deficiency of prior art, a kind of determined curative effect is provided, effective ingredient is easy to quantize, and quality is easy to control, the Chinese medicine ligroin extraction of the control glycolipid metabolism disorder that dose is little.

Another object of the present invention provides the preparation method of above-mentioned ligroin extraction.

The present invention is achieved through the following technical solutions above-mentioned purpose:

A kind of Chinese medicine ligroin extraction of preventing and treating the glycolipid metabolism disorder, its effective ingredient is ceryl alcohol, cupreol, n-hexacosanoic acid, atractylenolide, oleanolic acid, berberine, jateorhizine, salvianolic acid B, cyclolignocerane, 9,12-octadecadienoic acid, 5,7-dimethoxy coumarin, ginsenoside Rb1.

Above-mentioned effective ingredient ceryl alcohol, cupreol, n-hexacosanoic acid, atractylenolide, oleanolic acid, berberine, jateorhizine, salvianolic acid B, cyclolignocerane, 9,12-octadecadienoic acid, 5,7-dimethoxy coumarin, ginsenoside Rb1's part by weight are 1～5:1～8:1～8:1～5:1～8:1～8:1～5:1～5:1～5:1～5:1～5:1～6.

The preparation method of above-mentioned Chinese medicine ligroin extraction, adopt the prescription of the medicine " the loyal art of compound recipe is transferred fat side " (being called for short FTZ) of patent 200410051250.4 disclosed treatment hyperlipemias, each compositions in weight percentage is represented: Fructus Ligustri Lucidi 15～35%, Radix Cirsii Japonici 10～25%, the Rhizoma Atractylodis Macrocephalae 5～20%, Radix Salviae Miltiorrhizae 5～20%, the Cortex Eucommiae 5～15%, Fructus Citri Sarcodactylis 5～15%, preferred ingredient is: Fructus Ligustri Lucidi 15～35%, Radix Cirsii Japonici 10～25%, the Rhizoma Atractylodis Macrocephalae 5～15%, Radix Salviae Miltiorrhizae 5～15%, the Cortex Eucommiae 5～15%, Fructus Citri Sarcodactylis 5～15%, Radix Notoginseng 5～15%, Rhizoma Coptidis 3～10%.With 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 process C _1-3After alcohol extraction and/or water are carried, merge total extract, obtain preventing and treating the Chinese medicine ligroin extraction of glycolipid metabolism disorder again with the petroleum ether extraction total extract.C wherein _1-3Alcohol is methanol, ethanol or propanol.

Said method may further comprise the steps:

(1) 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 step (1) obtained adds 0.5～15 times, and the petroleum ether of preferred 6～10 times of volumes extracts 1～5 time, and preferred 3～4 times, combining extraction liquid, cold drying obtains preventing and treating the Chinese medicine ligroin extraction of glycolipid metabolism disorder.

C described in the step (1) wherein _1-3The ethanol extraction 1～5 time of 30～95 volume % is preferably used in alcohol extraction, and preferred 2～4 times, each C that extracts _1-3Pure volume be 1～15 times of quality of medicinal material, preferred 6～10 times, each extraction time is 5 min～5 h, preferred 2～3 h.

It is to boil 1～5 time with decocting that water described in the step (1) is carried, and preferred 2～4 times, the volume of each water is 1～15 times of quality of medicinal material, and preferred 6～12 times, each decocting time is 5 min～5 h, preferred 2～3 h.

Concentrate the concentrated solution volume be meant after concentrating described in the step (1) and be 0.2～5 times of quality of medicinal material, preferred 1～2 times.

The ligroin extraction that obtains is according to acceptable carrier on the pharmaceutics, use general formulation method, be processed into oral formulations or ejection preparation, as the form of tablet, capsule, powder, pill, powder, granule, crystal, solution, extractum, outstanding agent, soup, syrup, elixir, tea, wet goods.The pharmaceutical preparation or the health food that are used for preparation prevention or the disorderly relevant disease of treatment glycolipid metabolism relevant diseases such as (hyperlipemia, diabetes, metabolic syndrome) animal are atherosis.

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 compound of patent 200410051250.4, obtain effective extract part, 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.Animal vivo test shows that this compound recipe ligroin extraction has obvious blood fat reducing, blood sugar reducing function to experimental hyperlipidemia, hyperglycemia, and the effect of energy cholesterol reducing, improve lipid metabolism, prevent and treat the arteriosclerosis effect, and do not observe apparent side effect, increased the safety of medication.

Herbal mixture ligroin extraction provided by the invention has further been expanded the range of application that the loyal art of compound recipe is transferred fat side's effective component group, provides a kind of new way for preventing or treating glycolipid metabolism related diseases.The medicine of the present invention and the disorderly relevant disease of similar control glycolipid metabolism fat relatively, determined curative effect, the no obvious toxic-side effects of safety is used separately or with other medicines are composite good prospects for application is arranged all.

The Chinese medicine ligroin extraction of our preparation is compared with former side and is had the following advantages:

(1) dose is reduced to 100mg by original 6g extract, is reduced to original 1/60;

(2) content of effective ingredient improves about 60 times;

(3) ligroin extraction good stability provided by the invention, can prepare multiple controlled release formulations for oral administration, widen the loyal art of compound recipe greatly and transferred the application of fat side in field of pharmaceutical preparations, significantly improved the bioavailability that the loyal art of compound recipe is transferred fat side's effective ingredient, the convenient use;

(4) animal vivo test shows, extract of the present invention has obvious effect for reducing fat to experimental hyperlipidemia, while energy cholesterol reducing, improve lipid metabolism, simultaneously experimental hyperglycemia disease there is obvious blood sugar reducing function, improve the blood glucose metabolism, and do not observe apparent side effect, increased the safety of medication.For the drug effect of the hypercholesterolemia of high fat diet hyperlipemia, this ligroin extraction part index number is better than former side, has reached the purpose that reduces dose and do not reduce drug effect.

Description of drawings

Fig. 1: embodiment 3 herbal mixture ligroin extraction HPLC chromatograms.

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 embodiment 1 Chinese medicine compound total extract

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 2:1:1:1:1:2:1:1) are broken into coarse powder, distinguish reflux, extract, 3 times with 10 times, 8 times of medical material amount and 6 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 3 times all with the water of 12 times, 10 times and 8 times amounts in the side, and each 2 h merge 3 times the water extract, and vacuum decompression is concentrated into 1 g medical material/ml extracting solution, water body partly and the front alcohol extraction partly merge and obtain the Chinese medicine compound total extract.

The preparation at the Petroleum ether extraction position of embodiment 2 compound recipe total extracts

With the petroleum ether extraction of 10 times of volumes of compound recipe total extract 1 time, separate petroleum ether layer; With 8,6 times of volume petroleum ether extractiones each 1 time, separate to obtain petroleum ether layer again, merges 3 times petroleum ether extraction liquid, the recovery petroleum ether, again in 0.08MPa, 65 ℃ of vacuum dryings obtain ligroin extraction (FTZEE);

According to total inventory of Chinese medicine compound and the amount that obtains extract part, calculate the yield of extract.The yield of ligroin extraction gets ligroin extraction 3.0～5.0g) at 0.3%～0.5%(1kg crude drug.

The chemical composition analysis at the Petroleum ether extraction position of embodiment 3 compound recipe total extracts

Embodiment 2 compound recipe ligroin extractions (FTZEE) are analyzed through HPLC, get Fig. 1 sample finger printing.

FTZ petroleum ether part UPLC chromatographic condition: Agilent HC-C ₁₈Chromatographic column (4.6mm * 250mm, 1 μ m), guard column phenomenex KJO-4282-C ₁₈Post; Mobile phase adopts (B) binary gradient system of acetonitrile (A)-water (containing 0.25% glacial acetic acid+0.13% triethylamine).Gradient elution: 2 ~ 5% A (0 ~ 5 min), 5% ~ 20% A (5 ~ 35 min), 20% A (35 ~ 45 min), 20% ~ 35% A (45 ~ 70 min), 35% ~ 80% A (70 ~ 85 min), 80% ~ 100%A (85 ~ 87 min), 100%A (87 ~ 89 min).Detecting wavelength is 254 nm, 300 nm and 330 nm, and column temperature is 30 ℃, and flow velocity is 0.2 mL/min, sample size 10 μ L.

The qualitative analysis: the FTZ petroleum ether part mainly contains ceryl alcohol, cupreol, n-hexacosanoic acid, atractylenolide, oleanolic acid, berberine, jateorhizine, salvianolic acid B, cyclolignocerane, 9,12-octadecadienoic acid, 5, compositions such as 7-dimethoxy coumarin, ginsenoside Rb1, its UPLC-MS finger printing is seen accompanying drawing 1, and wherein the part by weight of each composition is 1～5:1～8:1～8:1～5:1～8:1～8:1～5:1～5:1～5:1～5:1～5:1～6.

The preparation method of embodiment 4 extract capsules

Extract makes up according to following weight portion: 5 parts of embodiment 2 ligroin extractions, 5 parts of starch, 5 parts of carboxymethyl celluloses.With each extract of aforementioned proportion, the starch mix homogeneously of ligroin extraction and equivalent mixes with remaining starch again, mixes with carboxymethyl cellulose at last, dry granulation behind three's mix homogeneously, behind the drying under reduced pressure, granulate, fill No. 3 capsule, polishing, quality inspection packing.Every contains FTZEE 30 mg.Taking dose is 3 times on the one, each 1.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 FTZEE capsule HPLC assay result

The capsule that makes is used to carry out the clinical treatment hyperlipemia, every day 3 times, each 1, serve on 60 days, patient's blood fat T-CHOL, triglyceride have obvious decline after the treatment of treatment group as a result, and the blood fat reducing total effective rate is 90.5%, and relatively there were significant differences (P＜0.01) with matched group.

The preparation method of embodiment 5 extract sheets

Add starch 5 kg with ligroin extraction 5 kg that prepare among the embodiment 2, carboxymethyl cellulose 5kg, mixing adds 75% ethanol and granulates, 80 ℃ of dryings, it is an amount of to add magnesium stearate, tabletting, the bag film-coat makes about 1,000,000 of FTZEE tablet, and every contains FTZEE 50 mg.Taking dose is 2 times on the one, each 1.The HPLC method is measured above-mentioned tablet, and each active constituent content measuring the results are shown in Table 2 in every:

Table 2 FTZEE sheet HPLC assay result

Adopt tablet to carry out the clinical treatment hyperlipemia, every day 2 times, each 1, serve on 60 days, patient's blood fat T-CHOL, triglyceride have obvious decline after the treatment of treatment group as a result, and the blood fat reducing total effective rate is 93.3%, and relatively there were significant differences (P＜0.01) with matched group.

The preparation method of embodiment 6 extract honeyed pill

Preparation FTZEE 5.0 kg, mixing makes the FTZEE honeyed pill.

Above FTZEE adds starch 3 kg, dextrin 3.0 kg, and the general ball of Mel 8. 0 kg becomes water-honeyed pill, and the dry ball of dry 15 kg is packaged into the pill of 100,000 granules, promptly gets 0.15g/ grain (every contains FTZEE50 mg).Oral dose: one time 1 granule, 2 times on the one.

The honeyed pill that makes is used to carry out the clinical treatment hyperlipemia, every day 2 times, and each 1, serve on 60 days, patient's blood fat T-CHOL, triglyceride have obvious decline after the treatment of treatment group as a result, and relatively there were significant differences (P＜0.01) with matched group.The blood fat reducing total effective rate is 91.5%.

Embodiment 7 acute toxicities and long term toxicity test

(1) acute toxicity test in mice: 120 19～21g NIH mices (male and female half and half) are divided into 6 groups, and 20 every group, FTZEE(embodiment 2 methods with five kinds of various dose prepare respectively) oral administration gavage ( P.o.) administration, normal saline (NS) negative control.Observe the reaction of mice and write down the mortality of every group of mice, adopt the linear regression software of Microsoft Excel, draw straight line, calculate LD according to log10 dose and empirical probability ₅₀FTZEE is to the LD of white mice oral administration gavage ₅₀Be respectively 450～751 mg/kg(and be equivalent to 300～500 times of the clinical consumption of people).

(2) rat long term toxicity test: get 120 male and female half and half of Wistar rat and be divided into 4 groups, 30 every group, respectively P.o.Give 45 mg/kg, 90 mg/kg, 180 mg/kg herbal mixture ligroin extractions, the NS negative control P.o.Normal saline.Successive administration 180 days, it is no abnormal that three dosage groups (be equivalent to the clinical consumption of people 30～120 times) rat there is no obvious hemogram, cardiopulmonary hepatic and renal function and nervous system, do not see that also vitals such as the heart, lung, spleen, stomach, brain, intestinal have obvious pathological change.

Conclusion: herbal mixture ligroin extraction toxicity of the present invention is very low, safe.

Embodiment 8 FTZEE are to the metabolic influence of diet high blood lipid model animal lipid

1. experiment material

Medicine and reagent: FTZEE, preparation method is with embodiment 2; The loyal art of compound recipe is transferred fat formula extraction FTZ; T-CHOL, triglyceride, determine cholesterol with high density lipoprotein test kit.

Animal: cleaning level SD rat, body constitution amount (180～225 g), economizing Experimental Animal Center by Nanfang Medical Univ provides; Purebred new zealand female rabbit, body constitution amount (1.80～2.20 kg) are provided by Guangdong Province's Experimental Animal Center.

2. experimental technique

2.1 influence to the blood fat of diet high blood lipid model rat

The FTZEE that observes various dose is to hyperlipemia whole animal Serum TC, TG, LDL-C, the influence of HDL-C.

(the loyal art of compound recipe is transferred the effect of fat side to diet hyperlipidemia rats hepatic lipase for Guo Jiao, Bei Weijian etc., Chinese crude drug, 2009,31 (4): 582～585) duplicate and confirm the success of hyperlipemia rat animal model to press the bibliographical information method.

With the animal random packet: be divided into 8 groups at random after 96 rats numberings are weighed, 12 every group, set 1. blank normal control group; 2. hyperlipidemia model group; 3. FTZ organizes; 4. lovastatin group; 5. fenofibrate group; 6. 7. 8. be followed successively by each experimental group of the high, medium and low dosage of FTZEE (15,30,60 mg/kg).Except that the normal control group is freely absorbed the normal diet, every morning gives high lipoprotein emulsion by 10 ml/kg amount filling stomach to all the other groups on this basis respectively, give corresponding medicine behind 4 h, wherein normal group, model group give the equivalent normal saline, in 4 weeks of successive administration, fasting 1 d (can't help water) gets blood from the eye socket venous plexus earlier behind the 2nd day last administration in early morning 1 h, centrifugal, separation of serum are measured TC, TG, HDL-C, LDL-C respectively.

Statistical method: data are represented with x ± s, carry out multiple comparisons between variance analysis and mean with SSP10.0 software.

3 results

3.1 influence to the blood fat of diet high blood lipid model rat

Result of study shows, FTZEE can obviously reduce T-CHOL (TC) in the diet high blood lipid model mice serum, triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C), obviously high density lipoprotein increasing (HDL-C) has the effect of regulating the blood ester, sees Table 3:

Table 3 FTZEE is to the influence of the blood fat of diet high blood lipid model rat

Annotate: compare * * P＜0.01 with matched group; Compare with high fat, F=3.82, ^▲P＜0.05, ^{▲ ▲}P＜0.01.

FTZ and FTZEE have obvious effect for reducing blood fat to diet high blood lipid model rat, and present certain dose-effect relationship.Prevent and treat the effect of experimental metabolism disorder of blood lipid, its mechanism of action may comprise:

(1) FTZEE of the present invention shows through serial animal experiment study, this side's blood fat reducing is evident in efficacy, have prevention and treatment hyperlipidemia dual function simultaneously concurrently, decapacitation influences the lipid metabolism key enzyme, effectively outside the blood lipid regulation, also have multiple actions such as the hemorheology of improvement, antioxidation, atherosclerosis.

(2) result of study proves, and FTZEE can strengthen CYP7A1 and HL enzymatic activity in the hyperlipidemia model rat blood serum greatly.

Embodiment 9 FTZEE express and active influence hepatocyte blood lipid metabolism key gene

Investigating ligroin extraction of the present invention expresses and active influence the blood lipid metabolism key gene.

1. method

1.1 to L-O2 hepatocyte cholesterol 7-α-hydroxylase (CYP7A1) and the active influence of hepatic lipase (HL)

Cultivate L-O2 people's normal liver cell routinely, treat to add respectively behind the cell fusion FTZEE(embodiment 2 methods preparation of various variable concentrations) and FTZ, after continuing to cultivate 24 h, collecting cell is made cell homogenates, use the HL detection kit, adopt colorimetry to detect the HL activity, study the influence of above-mentioned variable concentrations FTZEE, relatively HL enzymatic activity in the hepatocyte under the variable concentrations FTZEE effect the HL1 enzymatic activity.

FTZEE and liver LO2 cell are educated altogether and are cultivated 24 h after the time, collect hepatocyte, (see Hylemon for details, P. B. by reported method such as Hylemon; Studer, E. J.; Pandak, W. M. et al 1989. Simultaneous measurement of cholesterol 7 alpha-hydroxylase activity by reverse-phase high-performance liquid chromatography using both endogenous and exogenous [4-14C] cholesterol as substrate. Analysic Biochemistry 182 (2): 212-216) preparation hepatocyte microsome, use the CYP7A1 detection kit, adopt the HPLC method to detect CYP7A1 activity (Hylemon et al., 1989), research FTZEE compares CYP7A1 activity change in the hepatocyte under the variable concentrations FTZEE effect to the influence of CYP7A1 enzymatic activity.

1.2 the HMGCoA reductase activity is suppressed experiment

Carry out external HMGCoA reductase enzymatic activity with above-mentioned FTZEE and suppress experiment, measure and calculate the IC that they suppress the HMGCoA reductase ₅₀, and contrast with pravastatin.

1.3 influence to L-O2 hepatocyte CYP7A1 and HL gene expression

Cultivate L-O2 people's normal liver cell routinely, treat to add the FTZEE of 1.0 μ g/ml and 5.0 μ g/ml respectively behind the cell fusion and the FTZ of 25.0 μ g/ml is three groups, after continuing to cultivate 24 h, collecting cell, extract total RNA in the hepatocyte with Trigol reagent, the RT-PCR method is measured CYP7A1 and HL enzyme gene expression situation in the hepatocyte, observes the influence of FTZEE to CYP7A1 in the hepatocyte and HL enzyme gene expression

2 statistical method

Data are represented with means standard deviation, carry out significance test with variance analysis method.

3 experimental results

3.1 influence to the hepatocyte hepatic lipase

Various dose FTZEE and FTZ handle the active significantly increase (P＜0.01) of back hepatocyte hepatic lipase, and obvious dose dependent is arranged, and the results are shown in Table 4.Illustrate that Chinese medicine ligroin extraction of the present invention (FTZEE) has the effect of significantly improving to hepatocyte hepatic lipase activity, help strengthening falling clearly of lipoprotein and reduce blood triglyceride and cholesterol.

3.2 influence to hepatocyte CYP7A1

Various dose FTZEE and FTZ handle the active significantly increase (P＜0.01) of back CYP7A1, and the enhancing of obvious dose dependent is arranged, and the results are shown in Table 4.Illustrate that Chinese medicine ligroin extraction of the present invention (FTZEE) has the effect of significantly improving to hepatocyte CYP7A1 activity, help promoting that cholesterol changes into bile acid, and drain, thereby reduce cholesterolemia.

Table 4 different pharmaceutical is to L-O2 cell CYP7A1 and the active influence of HL

Annotate: compare * P＜0.05 with the normal control group; * P＜0.01.

3.4 inhibitory action to the HMG-CoA reductase activity

Make HMG-CoA reductase solution with the rat hepatocyte microsome, the determination of activity of HMG-CoA reductase then adopts literature method to carry out (Kleinsek, D.A., Ranganathan, S. and Porter, J.W., 1977. Purification of 3-hydroxy-3-methylglutaryl-coenzyme A reductase from rat liver. Proceedings of the National Academy of Sciences USA 74,1431-1435) measure enzymatic activity under each medicine prescribed concentration respectively, obtain the IC of each medicine such as FTZEE HMG-CoA reductase enzymatic activity ₅₀(μ g/ml).

The results are shown in Table 5, the result shows that FTZEE can obviously reduce rat HMG-CoA reductase enzymatic activity, and is the dose dependent relation.FTZEE can suppress the activity of HMG-CoA reductase, not only hinders the synthetic of the synthetic critical materials HMG-CoA of cholesterol, and suppresses the biosynthesis of cholesterol, produces the cholesterol reducing effect.

Table 5 FTZEE is right-IC of HMG-CoA reductase activity ₅₀(μ g/ml)

FTZEE suppresses liver HMG-COA R and reduces enzymatic activity, and it is synthetic to suppress liver TC, and liver TC content significantly reduces.

Embodiment 10 Chinese medicine ligroin extractions are to the influence to B cell

1.1 method

1.1.1 influence to B cell

Cultivate rat Langerhans islet B cell routinely, treat to add 200 μ M H respectively behind the cell fusion ₂O ₂Educate 1 h altogether, add the FTZEE(embodiment 2 methods preparation of various variable concentrations then) and FTZ, continue to cultivate 24 h after the time, collecting cell is made cell homogenates, with superoxide 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 FTZEE B cell SOD, CAT and GSH-Px activity and the active influence of GSH content.

1.1.2 influence to Bcl-2, PPAR-a and Bax in the B cell and Fas gene expression

Handle the same 1.1.1 of B cell method, extract total RNA in the B cell with Trizol reagent, the RT-PCR method is measured Bcl-2 and Bax and Fas expression conditions in the B cell, observes the influence of FTZEE to Bcl-2, PPAR-a and Bax in the B cell and Fas gene expression.

1.2 statistical method: data are represented with means standard deviation, carry out significance test with variance analysis method.

1.3 experimental result

1.3.1 influence to B cell SOD, CAT and GSH-Px activity and GSH content

Various dose FTZEE 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 6.Illustrate that FTZEE of the present invention 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 6 different pharmaceutical is to the influence of oxidative stress damage B cell SOD, CAT and GSH-Px activity and GSH content

Annotate: (1) is compared 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 the fat formula extraction, FTZEE: the loyal art of compound recipe is transferred fat ashlar oil ether extract, BBR: berberine hydrochloride.

1.3.2 influence to Bcl-2 in the B cell and Bax and Fas gene expression

Bcl-2 and Bax and Fas gene expression are expressed normal in the normal group B cell; Bcl-2 gene expression significantly increases (P＜0.01) in the B cell of various dose FTZEE processing back; and Bax and Fas gene expression significantly reduce (P＜0.01); illustrate that gene expression has obvious facilitation (P＜0.01) to FTZEE to Bcl-2; 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 7.

Table 7 different pharmaceutical is to the influence of Bcl-2, PPAR-a and Bax in the B cell and Fas gene expression

Annotate: (1) is compared 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 the fat formula extraction, FTZEE: the loyal art of compound recipe is transferred fat ashlar oil ether extract, BBR: berberine hydrochloride.

Embodiment 11 Chinese medicine ligroin extractions 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 5 groups at random by sex: blank group, glibenclamide group, compound recipe total extract group, 1,2,3 groups of ligroin extraction groups (preparation of embodiment 2 methods), 10 every group.

Experimental technique: behind animal fasting 12 h, get blood from the eye socket venous plexus, centrifugal 10 min of blood sample 3000 rpm, separation of serum by the operation of glucose kit description, is measured blood sugar content.Get and irritate stomach behind the blood immediately and give to corresponding for 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 8.

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 8 Chinese medicine ligroin extraction is to the effect of normal mouse blood sugar (n=10 x ± s)

With matched group ratio, * P＜0.05.

Experimental result shows, with the normal mouse ratio, administration after 3,6,9 hours each group of positive control drug glibenclamide group, total extract group and Petroleum ether extraction blood glucose of normal mouse is had significant reduction effect, each organize administration after 6 hours hypoglycemic effect the most remarkable.

Embodiment 12 Chinese medicine ligroin extractions (FTZEE) are to the influence of the blood glucose of diabetic mice

Medicine and reagent: FTZEE(Chinese medicine ligroin extraction, the preparation of embodiment 2 methods); Blood glucose test kit (the safe reagent company limited of Beijing northization); T-CHOL, triglyceride, determine cholesterol with high density lipoprotein test kit; Streptozotocin (STZ), U.S. Sigma company product; The peace of quenching one's thirst 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.

To the blood glucose of streptozotocin (STZ) type diabetic mice and the influence of blood fat

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 FTZEE (30,60,120 mg/kg), FTZ2 group (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.

The 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 big low dose of FTZEE (120,60mg/kg) group, FTZ 250 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 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.

The 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, every group 10, the high sugared model group of normal control group, glucose, the big low dose of FTZEE (120,60mg/kg) group, FTZ 250 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 glucose group give equal-volume normal saline (NS), successive administration 7 days, behind last administration 2 h, matched group lumbar injection (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 9, table 10, table 11.

Table 9. FTZEE 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.

The influence of the blood glucose of the hyperglycemia mice that table 10. FTZEE causes epinephrine (n=10, x ± s)

Annotate: compare with normal group, ^*P＜0.05, ^*P＜0.01; Before treating, compare, ^{▲ ▲}P＜0.01.

The influence of the blood glucose of the hyperglycemia mice that table 11. FTZEE causes glucose (n=10, x ± s)

Annotate: compare with normal group, ^*P＜0.01; With comparing before the treatment ^▲P＜0.05, ^{▲ ▲}P＜0.01.

Embodiment 13 FTZEE bring 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 (NS) 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 big low dose of stomach FTZEE (60,120 mg/kg, the preparation of embodiment 2 methods) 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 0.5,2 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 12, table 13.

Table 12 FTZEE brings out influence (n=10, the x ± s) of the plain resistance of mouse islets to hydrocortisone

Table 13 FTZEE brings out influence (n=10, the x ± s) of the plain resistance of mouse islets to hydrocortisone

Annotate: compare with normal group, ^*P＜0.01; Same hydrocortisone+NS group is compared ^▲P＜0.05, ^{▲ ▲}P＜0.01.

Embodiment 14 FTZEE are 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 12 h, 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, pH 4.5, 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 72 h.Diabetes rat is divided into diabetic model group, the big small dose group of FTZEE (preparation of embodiment 2 methods) at random.Big agent group by people and rat dose,equivalent than mg/5 ml/ kg administration every days 40, little dose of group mg/5 ml/kg every days 20, 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 14, 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 FTZEE has remarkable hypoglycemic activity.

Table 14 FTZEE 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＜0.05, ^{▲ ▲}P＜0.01.

Influence to serum insulin the results are shown in Table 15, model group and normal group are relatively, 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 FTZEE to Ins after the big or small dosage treatment.

Table 15 FTZEE 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.With comparing before the modeling, ^△P＜0.05, ^{△ △}P＜0.01; Compare with model group ^▲P＜0.05, ^{▲ ▲}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; Small dose group islets of langerhans number and island inner cell number increase, and form is rule comparatively, the small part karyopycnosis.This explanation FTZEE has significant protective effect to the islet cells tissue of STZ MODEL DAMAGE, and can promote secretion of insulin.

Embodiment 15 prevents and/or treats the clinical verification of hyperglycemia/diabetes

FTZEE sheet (Chinese medicine ligroin extraction sheet, the preparation of embodiment 5 methods) treatment diabetes 60 examples are analyzed

Test is with reference to Zheng Xiao cornel chief editor, new Chinese medicine clinical research guideline (try), Chinese Medicine science and technology publishing house, 2002 the 1st edition, 233-237: new Chinese medicine is treated the clinical research guideline of diabetes and is carried out.

1. clinical data

60 routine patients all are the non-insulin-depending type inpatient of making a definite diagnosis according to diagnostic criteria in 1993, wherein, and outpatient service 40 examples, 20 examples of being in hospital; Be divided into two groups at random, treatment is organized in 30 examples, male 16 examples, women 14 examples; 39 ~ 65 years old age, year mean age (51.3 ± 8.6).Diabetic duration 3 ~ 15 years, average course of disease (6.8 ± 2.9) year; Merge hyperlipidemia person's 19 examples, coronary disease patient 10 examples, hyperpietic's 18 examples; In matched group 30 examples, male 15 examples, women 15 examples; 38 ~ 65 years old age; Year mean age (51.5 ± 6.8); Diabetic duration 2.5 ~ 16 years, average course of disease (6.9 ± 3.1) year; Merge hyperlipidemia person's 20 examples, coronary disease patient 9 examples, hyperpietic's 17 examples.Learning processing aspect sex, age, the course of disease, the complication by statistics for two groups, and no significant difference (P〉0.05), have comparability.

2. diagnostic criteria

Western medicine diagnose standard (according to WHO expert consulting report in 1999): fasting glucose (FPG) 〉=7.0 mmol/L and 2 hours after the meal blood glucose (2 HPG) 〉=7.0 mmol/L, random blood sugar 〉=11.1 mmol/L.

The high pressure Herba Wedeliae Wallichii according to hypertension alliance of World Health Organization (WHO) in 1999/international about hypertensive classification by stages method formulate that (hypertension alliance of World Health Organization (WHO) in 1999/international is about the hypertension therapeutic guide, hypertension magazine, 1999,7 (2): 9).

The clinical research guideline (1993) of tcm diagnosis standard reference Ministry of Health of the People's Republic of China traditional Chinese medical science new drug treatment diabetes (diabetes): all have or dry mouth and throat, fatigue and weakness, polyorexia, thirst and liking drink, the lazy speech of breathing hard, dysphoria with feverish sensation in the chest palms and soles, palpitation and insomnia, pain in chest and hypochondrium, dark coloured urine constipation, red tongue or dim red, pale purple or the petechia ecchymosis arranged, deep-thready pulse or string are puckery, and Chinese medical discrimination is deficiency of both QI and YIN, blood stasis venation card person.

3. Therapeutic Method

The treatment group is taken Chinese medicine ligroin extraction tablet (being called for short the FTZEE sheet), and each 1, every day 2 times, 2 months was 1 course of treatment, other hypoglycemic medicine of stopping using therebetween.

Matched group is taked to quench one's thirst and is pacified capsule (0.4g/ grain, Tonghua Baishan Pharmaceutical Ltd produces) treatment, and each 3, every day 3 times, 2 months was 1 course of treatment, the treatment type 2 diabetes mellitus.

Two groups of patients all carry out diabetes education, diet control, moderate exercise.All case observation is 2 months.

4. observation index

2,4,6 weeks detected blood pressure simultaneously during measuring blood pressure, fasting glucose (FBG) and 2 hours after the meal blood glucose (2 HFBG) (glucose oxidase method), glycosylated hemoglobin (HbALc), hepatic and renal function, routine urinalysis, the treatment before and after the treatment.

5. clinical efficacy

Clinical research guideline (1993) criterion of therapeutical effect with reference to Ministry of Health of the People's Republic of China's traditional Chinese medical science new drug treatment diabetes (diabetes) is evaluated.Produce effects: transference cure, lab testing are repeatedly normal; Effectively: cardinal symptom and relevant lab testing have improvement; Invalid: symptom and relevant lab testing no change.

Blood glucose, hemorheology situation of change are shown in table 16 and 17 before and after the patient.

Blood glucose, HbALc are relatively before and after the table 16 liang group treatment

Annotate: relatively preceding with the treatment of this group, ^★P＜0.05, ^{★ ★}P＜0.01.

By table 16 as seen, behind two groups of patient treatments on an empty stomach and 2 hours after the meal blood glucose, glycosylated hemoglobin obvious decline is all arranged; Relatively there were significant differences (P＜0.01) with matched group, and similar effect is also arranged after the treatment of control group, illustrate that FTZEE treats type 2 diabetes mellitus and significant curative effect arranged aspect blood glucose, the urine protein improving.

Table 17 illustrates the curative effect statistics of FTZEE sheet to diabetes, metabolism syndrome, shows that the FTZEE sheet has hypoglycemic curative effect to diabetic.

Table 17FTZEE sheet is to the curative effect statistics of diabetes, metabolism syndrome

Annotate: compare with the diabetes pill group, ^*P＜0.05.

Therefore, Chinese medicine ligroin extraction of the present invention (FTZEE) sheet is 90.0% to the total effective rate of diabetes, and produce effects 60.0% is slightly higher than 87% total effective rate of present medicine commonly used, and does not observe apparent side effect.

Conclusion: FTZEE confirms that through the modern Chinese medicine pharmacological research tool promotes insulin secretion, 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-a 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, the Chinese medicine ligroin extraction FTZEE that the present invention prevents and treats the glycolipid metabolism disorder has significant blood sugar lowering, improves the blood glucose metabolism, and treatment hyperglycemia and diabetes blood glucose metabolism disorder relevant disease clinical efficacy are definite.

Claims (9)

1. Chinese medicine ligroin extraction of preventing and treating the glycolipid metabolism disorder, its effective ingredient is ceryl alcohol, cupreol, n-hexacosanoic acid, atractylenolide, oleanolic acid, berberine, jateorhizine, salvianolic acid B, cyclolignocerane, 9,12-octadecadienoic acid, 5,7-dimethoxy coumarin, ginsenoside Rb1.

2. Chinese medicine ligroin extraction according to claim 1, it is characterized in that ceryl alcohol, cupreol, n-hexacosanoic acid, atractylenolide, oleanolic acid, berberine, jateorhizine, salvianolic acid B, cyclolignocerane, 9,12-octadecadienoic acid, 5,7-dimethoxy coumarin, ginsenoside Rb1's part by weight are 1～5:1～8:1～8:1～5:1～8:1～8:1～5:1～5:1～5:1～5:1～5:1～6.

3. the preparation method of the described Chinese medicine ligroin extraction of claim 1 is through C with 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 _1-3After alcohol extraction and/or water are carried, merge total extract, obtain preventing and treating the Chinese medicine ligroin extraction of glycolipid metabolism disorder again with the petroleum ether extraction total extract.

4. method according to claim 3 is characterized in that may further comprise the steps:

(1) 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 step (1) obtained adds the petroleum ether of 0.5～15 times of volume, extracts 1～5 time, and combining extraction liquid, cold drying obtains preventing and treating the Chinese medicine ligroin extraction of glycolipid metabolism disorder.

5. preparation method according to claim 4 is characterized in that C described in the step (1) _1-3Alcohol extraction is the C that selects 30～95 volume % for use _1-3Alcohol extraction 1～5 time, each C that extracts _1-3The alcohol volume is 1～15 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 it is to boil 1～5 time with decocting that water described in the step (1) is carried, and the volume of each water is 1～15 times of quality of medicinal material, and each decocting time is 5 min～5 h.

7. preparation method according to claim 4, the concentrated solution volume after it is characterized in that concentrating in the step (1) are 0.2～5 times of quality of medicinal material.

8. preparation method according to claim 4, it is characterized in that be the ligroin extraction that will obtain according to acceptable carrier on the pharmaceutics, be processed into oral formulations or ejection preparation.

9. preparation method according to claim 8 is characterized in that described oral formulations is tablet, capsule, powder, pill, powder, granule, crystal, solution, extractum, outstanding agent, soup, syrup, elixir, tea or oil.

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