CN102935151B - Medicine composition for protecting liver and lowering transaminase and preparation method and application thereof - Google Patents

Abstract

The invention discloses a medicine composition for protecting liver and lowering transaminase. The medicine composition is the preparation prepared by the following raw medicines by weight: 7 to 13 parts of oriental wormwood, 0.2 to 0.4 part of bear gall powder, 4.2 to 7.8 parts of rhizoma atractylodis macrocephalae, 6 to 12 parts of rhizoma alismatis, 7 to 13 parts of hawthorn, 4.2 to 7.8 parts of radix bupleuri, and 4.2 to 7.8 parts of liquorice. The invention also provides a preparation method and application of medicine composition. By adopting the medicine composition provided by the invention, the steatotic hepatocyte can be obviously alleviated, the lipid aggregation in hepatic tissue can be reduced, the blood serum TC (Total Cholesterol), TG (Triglyceride) and LDL (Low Density Lipoprotein) can be reduced, and the HDL (High Density Lipoprotein) can be raised; and the medicine composition is high in activity of protecting liver, lowering transaminase and reducing lipid, can be effectively used for treating non-alcoholic fatty liver disease, so that a new choice is provided for clinical medication.

Description

Pharmaceutical composition of a kind of the liver protecting and ALT lowering and its production and use

Technical field

The present invention relates to pharmaceutical composition of a kind of the liver protecting and ALT lowering and its production and use.

Background technology

In recent years, irrational diet structure, be addicted to drink, sit less dynamic life style more, have the factors such as fat family history to cause the sickness rate of China's fatty liver to raise year by year, account for 10% of average population, Adult Onset leads and may reach 20% to 30%, fat, to be addicted to drink and can up to 50%-60% in diabetic population, wherein there is hepatic fibrosis in the patient of about 25%, and the patient evolution of 1.5-8.0% is liver cirrhosis.Current, Patients with Fatty Liver rises year by year, and presents the development trend that becomes younger.Therefore, support the medicament research and development of effectively treatment fatty liver, the treatment for the current fatty liver of China seems very urgent and necessary.

The traditional Chinese medical science is thought, its disease located in liver of fatty liver, gallbladder, spleen, stomach; Because the Different types of etiopathogenises such as meridians of damp and hot, expectorant is turbid, congestion etc. becomes silted up resistance liver causes, diet, obesity, quench one's thirst relevant with the generation of primary disease, as " the strange sick opinion of Plain Questions " meaning: " this fertile institute is sent out, and this person must count and eat sweet and refreshing and many fertilizer also.Overeating greasy food bringing about internal heat, over-eating the food with sweet flavor bringing about abdominal distension, therefore its gas overflow, transfer to and quenching one's thirst." " Ling Shu Miraculous Pivot or Divine Axis disorder of defensive QI " say: " cream person, many gas and skin is vertical slow, therefore can indulge abdomen and hang down fat ", add: " gas is slided less for fat person, its serum, therefore can not greatly ".ZHU Dan-xi proposes the viewpoint of " fertile white man's excessive phlegm wets " in " the ZHU Dan-xi method for the treatment of heart is wanted ", and Liu's interchannel proposes fertile people, and " the many stasis of space between skin and muscles, QI and blood is difficult to tonneau." " Zhang Shi doctor logical " point out: " people be addicted to drink, sick abdominal distention is as bucket, and this obtains damp-heat impairing the spleen." "guide to clinical practice with medical record" points out: " and but wet from interior survivor, must its people's fat meat and fine grain wine sweet wine excessive.”

The treatment of ancients to this disease proposes many effective methods."guide to clinical practice with medical record" is said: " assorted card hypochondriac pain; all belong to cloudy Liver Channel of fainting; be distributed in side of body rib with the liver pulse, therefore Zhong Jing Flos Inulae soup, interchannel Sichuan Chinaberry Powder and Mr. (Ye Shi) pungent temperature dredging collateral, the slow reason of liver emptyly, gentle logically to be mended, are pungently let out the methods such as Xuan Mi, all agent for the treatment of the liver work hypochondriac pain." " Jing Yue's complete work gathers " point out: " and control long-pending wanting, attack the suitable of benefit knowing, when in who slow who anxious in distinguish it.”

Fatty liver is divided into stagnation of QI due to depression of the liver, spleen deficiency of kidney-QI, turbid damp internal resistance, qi depression to blood stasis four kinds card shape by TCM more.Treatment adopts respectively the methods such as soothing liver and strengthening spleen, tonifying speen and tonifying kidney, damp eliminatingization are turbid, blood circulation promoting and blood stasis dispelling, achieve certain curative effect.

Summary of the invention

The object of the present invention is to provide pharmaceutical composition of a kind of the liver protecting and ALT lowering and its production and use.

The invention provides a kind of pharmaceutical composition of the liver protecting and ALT lowering, it is the preparation be prepared from by the crude drug of following weight proportion:

Herba Artemisiae Scopariae 7-13 part, Fel Ursi powder 0.2-0.4 part, Rhizoma Atractylodis Macrocephalae 4.2-7.8 part, Rhizoma Alismatis 6-12 part, Fructus Crataegi 7-13 part, Radix Bupleuri 4.2-7.8 part, Radix Glycyrrhizae 4.2-7.8 part.

Further, it is the preparation be prepared from by the crude drug of following weight proportion:

Herba Artemisiae Scopariae 10 parts, Fel Ursi powder 0.3 part, the Rhizoma Atractylodis Macrocephalae 6 parts, Rhizoma Alismatis 9 parts, Fructus Crataegi 10 parts, Radix Bupleuri 6 parts, 6 parts, Radix Glycyrrhizae.

Wherein, described Radix Bupleuri is Radix Bupleuri Bupleurum chinense DC..

Wherein, described preparation be with the water of each crude drug or extractive with organic solvent for active component, add the preparation that pharmaceutically conventional adjuvant or complementary composition are prepared from.

Further, described preparation is oral formulations, external preparation or ejection preparation.

Further, described oral formulations is solid preparation.

Further preferably, described oral formulations is granule, tablet, capsule, powder or pill.

Present invention also offers the preparation method of aforementioned pharmaceutical compositions, it comprises following operating procedure:

(1) according to consumption proportion weighting raw materials;

(2) Herba Artemisiae Scopariae, Rhizoma Alismatis, the Rhizoma Atractylodis Macrocephalae, Fructus Crataegi, Radix Bupleuri, Radix Glycyrrhizae is got, first extracting in water, ethanol is added again to alcohol content 60-90%v/v after extracting solution is concentrated, after precipitate with ethanol, get supernatant, reclaim ethanol, concentrate or after drying, add Fel Ursi powder and pharmaceutically conventional adjuvant or complementary composition and be prepared into preparation; Or

(3) a, get Fel Ursi powder, after beta-cyclodextrin inclusion compound, obtain Fel Ursi powder clathrate;

B, get Rhizoma Alismatis, Herba Artemisiae Scopariae, Radix Glycyrrhizae, add 70-90%v/v ethanol extraction, after extracting solution is concentrated, upper nonpolar or low pole macroporous adsorbent resin, after first washing removing impurity with water, then with 70-90%v/v ethanol for eluant, collects ethanol position, obtains eluent A;

C, get Radix Bupleuri, the Rhizoma Atractylodis Macrocephalae, Fructus Crataegi, extracting in water, after extracting solution is concentrated, upper nonpolar or low pole macroporous adsorbent resin, after first washing removing impurity with water, then with 70-90%v/v ethanol for eluant, collects ethanol position, obtains eluent B;

D, merge eluent A, B, after reclaiming ethanol, concentrated or dry, then add Fel Ursi powder clathrate and pharmaceutically conventional adjuvant or complementary composition and be prepared into preparation.

Wherein, in step (2), alcohol content is 65-80%v/v; In the b item of step (3), macroporous adsorbent resin is D101, and eluant is 70%v/v ethanol; In the c item of step (3), macroporous adsorbent resin is AB-8, and eluant is 70%v/v ethanol;

Wherein, described in step (2) or (3), drying is spraying dry, and drying condition is as follows: get thing to be dried, is first prepared into the liquid of density 1g/ml, then at inlet temperature 142 ~ 150 DEG C, leaving air temp 76 ~ 80 DEG C, carries out spraying dry; In a item of step (3), the amount ratio of Fel Ursi powder and beta-schardinger dextrin-is 1:70-90w/w, is that solvent carries out enclose with water.

Present invention also offers the purposes of above-mentioned pharmaceutical composition in the medicine of preparation treatment non-alcoholic fatty liver disease.

Further, described medicine is the medicine for the treatment of Primary nonalcoholic fatty liver disease.

The present invention also provides aforementioned pharmaceutical compositions preparing the liver protecting and ALT lowering or improving the purposes in the medicine of dyslipidemia.

In pharmaceutical composition of the present invention, Herba Artemisiae Scopariae is bitter, pungent, be slightly cold, and returns spleen, stomach, liver, gallbladder meridian, dampness removing, jaundice eliminating, removing toxic substances; And kind oozing is let out and diuresis, divides and disappears to walk to rush down damp-heat in middle-JIAO, make the wet orphan that reduces phlegm and internal heat, damp and hot from removing; " Records of Tradition Chinese and Western Medicine in Combination " cloud: " heat disappears strongly fragrant opening, and the road that bile enters small intestinal is also unseparated for the heat of kind liver and gall of spreading out, reason of holding concurrently liver and gall strongly fragrant "; Modern pharmacological research shows, Herba Artemisiae Scopariae has function of gallbladder promoting, protects the liver, effect for reducing blood fat.Therefore be monarch drug.Fel Ursi, bitter, cold, enter liver, gallbladder, heart channel, liver heat removing function of gallbladder promoting, heat-clearing and toxic substances removing, endogenous wind stopping relieving convulsion; " Newly Revised Canon of Materia Medica " cloud: Fel Ursi can " be treated seasonal epidemic pathogens intenseness of heat and become jaundice "; " book on Chinese herbal medicine covers and signs ": " control man, woman's seasonal epidemic pathogens heat is steamed, become jaundice "; Modern pharmacological research shows that Fel Ursi has and reduces triglyceride and cholesterol level in blood, effectively can prevent and treat hyperlipemia, Fructus Crataegi sour in the mouth, property are sweet, tepor, return spleen, stomach, Liver Channel, help digestion, eliminate indigestion, circulation of qi promoting, and can blood circulation promoting and blood stasis dispelling, can treat " lingering damp-heat with the passing of time; must qi depression to blood stasis be given birth to ", Compendium of Material Medica: " dissipating fluid-retention is eaten, and disappear meat Ji ， mass in the abdomen; phlegm retention feeling of fullness acid regurgitation, stagnant blood pain with distension "; Modern pharmacological research shows, Fructus Crataegi effectively can reduce blood fat, aid digestion, antioxidation, enhancing body immunity, is ministerial drug altogether.Rhizoma Alismatis is cold in nature, sweet light promoting diuresis to eliminate damp pathogen, and expelling the heat-evil is turbid, and " Mingyi Bielu " carries " tonifying deficiency five kinds of over strain, except the five internal organs feeling of fullness, plays cloudy gas, and antidiarrheal essence, quenches one's thirst, and drenches drop, the water by bladder, three Jiao "; Modern pharmacological research shows, Rhizoma Alismatis has the effects such as blood fat reducing, anti-fatty liver, atherosclerosis.The Rhizoma Atractylodis Macrocephalae is bitter, sweet, warm in nature, and return spleen, stomach warp, air making-up and spleen enlivening, dampness diuretic, matching with Rhizoma Alismatis forms golden deficient ZEXIE TANG, the card of Sheng in burnt deficiency of spleen-QI and stomach-QI, phlegm-damp in treatment, and prevents his medicine too bitter cold, clearing away heat-damp and promoting diuresis and imapirment of the spleen and stomach.Modern pharmacological research shows, the Rhizoma Atractylodis Macrocephalae has the effects such as hepatic cholagogic, diuresis, enhancing gastrointestinal peristalsis.Therefore be adjuvant drug.Radix Bupleuri bitter in the mouth, cold nature, dispersing the stagnated live-QI to relieve the stagnation of QI, makes irritability bar reach, and phlegm-damp is from removing.Compendium of Material Medica is recorded: " labor has five kinds of over strain, the disease being located in five ZANG organs.If labor has heat at liver, gallbladder, the heart and envelope, or the hot person of shaoyang channel, then Radix Bupleuri is that brothers faint the medicine that the moon, few sun must use ... ".And draw all medicines and enter liver; Modern pharmacological research shows Radix Bupleuri energy hepatic cholagogic, stabilizing cell membrane, has regulating action to protein, blood glucose, lipid metabolism, therefore for making medicine.Radix Glycyrrhizae is sweet flat, and QI invigorating is reduced phlegm, coordinating the actions of various ingredients in a prescription, also for making medicine.Full side's compatibility, using medicines of both cold and hot natures simultaneously, diuretic and non-impairment of YIN, heat clearing away and do not cut down stomach, make the clear wet profit of heat, the expectorant stasis of blood must be changed, and is the efficacious prescriptions for the treatment of liver gallbladder damp-heat type fatty liver.

Pharmaceutical composition provided by the invention, significantly can improve the gathering of lipid in hepatic cell fattydegeneration and liver, reduces serum TC, TG, LDL, raise HDL, there is good the liver protecting and ALT lowering Lipid-lowering activities, non-alcoholic fatty liver disease treatment can be effective to, for clinical application provides new selection.

Accompanying drawing explanation

Fig. 1: normal group HE dyeing hepatic tissue (lobus sinister) pathological section light microscopic view, amplification is 200 ×.

Fig. 2: Models of Fatty Liver group HE dyeing hepatic tissue (lobus sinister) pathological section light microscopic view, amplification is 200 ×.

Fig. 3: DONGBAO GANTAI matched group HE dyeing hepatic tissue (lobus sinister) pathological section light microscopic view, amplification is 200 ×.

Fig. 4: pharmaceutical composition low dose group HE dyeing hepatic tissue (lobus sinister) pathological section light microscopic view of the present invention, amplification is 200.

Fig. 5: dosage group HE dyeing hepatic tissue (lobus sinister) pathological section light microscopic view in pharmaceutical composition of the present invention, amplification is followed successively by 200.

Fig. 6: pharmaceutical composition high dose group HE dyeing hepatic tissue (lobus sinister) pathological section light microscopic view of the present invention, amplification is followed successively by 200.

Detailed description of the invention

The preparation of embodiment 1 pharmaceutical composition of the present invention

[prescription] Herba Artemisiae Scopariae 300g Rhizoma Alismatis 270g Rhizoma Atractylodis Macrocephalae 180g Fructus Crataegi 300g

Radix Bupleuri 180g Radix Glycyrrhizae 180g Fel Ursi powder 9g

[method for making] Herba Artemisiae Scopariae, Rhizoma Alismatis, the Rhizoma Atractylodis Macrocephalae, Fructus Crataegi, Radix Bupleuri, Radix Glycyrrhizae decoct with water secondary, each 1.5 hours, filter, merging filtrate, it is 1.20(50 DEG C that filtrate is concentrated into relative density) extractum, let cool, adding ethanol makes alcohol content be 70%, stirs, and leaves standstill 24h, make precipitation complete, centrifugal, Aspirate supernatant, 60 DEG C of decompression recycling ethanols, to without after alcohol taste, are concentrated into thick extractum and let cool, add dextrin, Fel Ursi powder 9g(is even prior to dextrin facing-up) make granule 1000g, drying, to obtain final product.

The preparation of embodiment 2 pharmaceutical composition of the present invention

Get Herba Artemisiae Scopariae 7g, Rhizoma Atractylodis Macrocephalae 7.8g, Rhizoma Alismatis 12g, Fructus Crataegi 13g, Radix Bupleuri 7.8g, Radix Glycyrrhizae 7.8g, decoct with water, merge decocting liquid, be concentrated into dry, add Fel Ursi powder 0.4g, and appropriate microcrystalline Cellulose, mixing, dry, encapsulated, obtain capsule.

The preparation of embodiment 3 pharmaceutical composition of the present invention

Get Herba Artemisiae Scopariae 13 parts, the Rhizoma Atractylodis Macrocephalae 4.2 parts, Rhizoma Alismatis 6 parts, Fructus Crataegi 7 parts, Radix Bupleuri 4.2 parts, 4.2 parts, Radix Glycyrrhizae, decoct with water, merge decocting liquid, after concentrated, add Fel Ursi powder 0.2 part, and appropriate amount of starch, granulate, dry, obtain granule.

The preparation of embodiment 4 pharmaceutical composition of the present invention

[prescription] Herba Artemisiae Scopariae 300g Rhizoma Alismatis 270g Rhizoma Atractylodis Macrocephalae 180g Fructus Crataegi 300g

Radix Bupleuri 180g Radix Glycyrrhizae 180g Fel Ursi powder 9g

[method for making] Herba Artemisiae Scopariae, Rhizoma Alismatis, the Rhizoma Atractylodis Macrocephalae, Fructus Crataegi, Radix Bupleuri, Radix Glycyrrhizae decoct with water secondary, each 1.5 hours, filter, merging filtrate, it is 1.20(50 DEG C that filtrate is concentrated into relative density) extractum, let cool, add ethanol and make alcohol content be 70%, stir, leave standstill 24h, make precipitation complete, centrifugal, Aspirate supernatant, 60 DEG C of decompression recycling ethanols are extremely without after alcohol taste, and it is 1.0g/ml that extracting solution is concentrated into density, inlet temperature 142 ~ 150 DEG C, carry out spraying dry under leaving air temp 76 ~ 80 DEG C of conditions, namely get dry extract;

Adopt the water of 12 times amount temperature more than 80 DEG C, 720g beta-schardinger dextrin-to be dissolved, (mixing speed is 400rmin in limit stirring ^-1), limit lowers the temperature, and slowly drips and add the Fel Ursi powder aqueous solution 180ml that concentration is 0.05g/ml 40 DEG C time, after stirring 80min, take out from thermostat water bath, put into the refrigerator cold preservation 24h of 4 DEG C, filter, residue washed with diethylether, and in 50 DEG C of dryings, obtain Fel Ursi powder clathrate;

Get dry cream, add Fel Ursi powder clathrate and appropriate dextrin, after mixing, make granule 1000g, dry, to obtain final product.

The preparation of embodiment 5 pharmaceutical composition of the present invention

[prescription] Herba Artemisiae Scopariae 300g Rhizoma Alismatis 270g Rhizoma Atractylodis Macrocephalae 180g Fructus Crataegi 300g

Radix Bupleuri 180g Radix Glycyrrhizae 180g Fel Ursi powder 9g

(1) extraction of Herba Artemisiae Scopariae, Rhizoma Alismatis, Radix Glycyrrhizae, purification

Get recipe quantity Herba Artemisiae Scopariae, Rhizoma Alismatis, Radix Glycyrrhizae, put in round-bottomed flask, with 8 times amount 80% alcohol reflux 2 times, be respectively 2h, decompression recycling ethanol at every turn, concentrated to put relative density be 1g/ml, for subsequent use.

The pretreatment of resin and dress post: resin is first used 95% soak with ethanol 24h, fully swelling, topple over and fall upper strata ethanol, wet method dress post, adding water not in white opacity with 95% ethanol elution to effluent, then be washed till without alcohol taste with distilled water, on resin, spread one deck absorbent cotton, with distilled water immersion, for subsequent use.

Purification by macroporous resin condition is: resin model: D101; Sample solution concentration: 0.5g/ml (crude drug amount); Leave standstill 2h, make absorption more complete; Eluant: 70% ethanol; Washing amount: 2BV; Eluting agent: 8BV; Flow velocity: 3BV/h.

(2) extraction of Radix Bupleuri, the Rhizoma Atractylodis Macrocephalae, Fructus Crataegi, purification

Get recipe quantity Radix Bupleuri, the Rhizoma Atractylodis Macrocephalae, Fructus Crataegi, put in round-bottomed flask, by 8 times of water gaging reflux, extract, 2 times, be respectively 2h, reclaim under reduced pressure at every turn, concentrated to put relative density be 1g/ml, for subsequent use.

The pretreatment of resin and dress post: resin is first used 95% soak with ethanol 24h, fully swelling, topple over and fall upper strata ethanol, wet method dress post, adding water not in white opacity with 95% ethanol elution to effluent, then be washed till without alcohol taste with distilled water, on resin, spread one deck absorbent cotton, with distilled water immersion, for subsequent use.

Purification by macroporous resin condition is: resin model: AB-8; Sample solution concentration: 0.5g/ml (crude drug amount); Leave standstill 2h, make absorption more complete; Eluant: 70% ethanol; Washing amount: 1BV; Eluting agent: 8BV; Flow velocity: 3BV/h.

(3) preparation of preparation

Get the eluent of step (1), (2) gained, merge, being evaporated to density is 2.0g/ml, carries out oven drying, obtain compound extract dried powder under 60 DEG C of conditions; Fel Ursi powder clathrate is prepared again by embodiment 1 method; Above-mentioned dried powder is mixed with Fel Ursi powder clathrate, adds appropriate dextrin, prepare granule.

The preparation of embodiment 6 pharmaceutical composition of the present invention

[prescription] Herba Artemisiae Scopariae 300g Rhizoma Alismatis 270g Rhizoma Atractylodis Macrocephalae 180g Fructus Crataegi 300g

Radix Bupleuri 180g Radix Glycyrrhizae 180g Fel Ursi powder 9g

[method for making] prepares Fel Ursi powder clathrate by embodiment 1 method; Get Herba Artemisiae Scopariae, Rhizoma Alismatis, the Rhizoma Atractylodis Macrocephalae, Fructus Crataegi, Radix Bupleuri, Radix Glycyrrhizae decoct with water secondary, each 1.5 hours, filter, merging filtrate, it is 1.20(50 DEG C that filtrate is concentrated into relative density) extractum, let cool, adding ethanol makes alcohol content be 70%, stirs, and leaves standstill 24h, make precipitation complete, centrifugal, Aspirate supernatant, 60 DEG C of decompression recycling ethanols are extremely without after alcohol taste, extracting solution is concentrated into the extractum that density is 2.0g/ml, lets cool; Add dextrin, Fel Ursi powder clathrate, make granule, then add appropriate magnesium stearate, after mixing, tabletting, obtains tablet.

The preparation of embodiment 7 pharmaceutical composition of the present invention

[prescription] Herba Artemisiae Scopariae 300g Rhizoma Alismatis 270g Rhizoma Atractylodis Macrocephalae 180g Fructus Crataegi 300g

Radix Bupleuri 180g Radix Glycyrrhizae 180g Fel Ursi powder 9g

[method for making] prepares Fel Ursi powder clathrate by embodiment 1 method, get Herba Artemisiae Scopariae, Rhizoma Alismatis, the Rhizoma Atractylodis Macrocephalae, Fructus Crataegi, Radix Bupleuri, Radix Glycyrrhizae decocts with water secondary, each 1.5 hours, filter, merging filtrate, it is 1.20(50 DEG C that filtrate is concentrated into relative density) extractum, let cool, adding ethanol makes alcohol content be 70%, stir, leave standstill 24h, make precipitation complete, centrifugal, Aspirate supernatant, 60 DEG C of decompression recycling ethanols are extremely without after alcohol taste, it is 2.0g/ml that extracting solution is concentrated into density, at power 5000W, pressure 0.08MPa, microwave drying 3h is carried out under temperature of charge 45-49 DEG C condition, obtain extractum solid, pulverize, cross 80 mesh sieves, for subsequent use, namely get dry extract powder.Add dextrin, Fel Ursi powder clathrate, make granule, dry, encapsulated, obtain capsule.

The beneficial effect of pharmaceutical composition of the present invention is described below by way of test example.

Test example 1 Drug therapy non-alcoholic fatty liver disease of the present invention animal effect experiment

1) animal grouping, modeling and administration

60 SD rats are divided into 6 groups at random, often organize 10, often organize equal male and female half and half, be namely divided into the high, medium and low dosage of normal group, Models of Fatty Liver group, DONGBAO GANTAI matched group and pharmaceutical composition of the present invention 3 treatment groups.Another 4 rats are incorporated to model group, use as sampling check modeling effect.

Except normal group, all the other are respectively organized rat and all replace normal feedstuff to feed 12 weeks with high lipid food.High lipid food proportioning: normal feedstuff 87.3%, Adeps Sus domestica 10%, cholesterol 2%, Fel Sus domestica salt 0.7%.Within 10th week, get each 2 of female, the great and mighty or powerful Mus of model group at random in modeling, observe modeling effect.

After modeling success the 10th week starts administration.While continuation modeling (preventing animal spontaneous recovery), normal group, Models of Fatty Liver group give 0.9% sodium chloride injection, DONGBAO GANTAI matched group is with administration 0.27g/kgd, and the basic, normal, high administration of pharmaceutical composition of the present invention is respectively: 0.5875g/kgd, 1.175g/kgd and 2.35g/kgd(dosage calculates by crude drug dosage) (being prepared by embodiment 1 method).All with the administration of solvent gavage mode, dosage is 2ml/250g.Above all in 9-11 point execution in the morning, every day 1 time, continuous 30 days.

2) observation index and test item

Gross examination of skeletal muscle

Observe each group of fresh liver color and luster, form, claim liver wet weights, measuring and calculating liver index.

Hepatic tissue HE dyes check pathological section

(1) cut 1cm × 1cm × 1cm piece of tissue in leftlobe of liver, preserve in 10% formalin and fix 24 hours.

(2) take out hepatic tissue blocking, carry out gradient alcohol dehydration, transparent, waxdip and paraffin embedding.

(3) section cycle type microtome transverse section serial section, thickness is 4 μm, drags for sheet, baking box is dried.

(4) tissue slice is with 100% dimethylbenzene dewaxing 10min × 2 time, and dehydrated alcohol, 95%, 80% graded ethanol soaks 5min × 1 time successively.

(5) tissue slice is washed return blue 15min, 5% hydrochloric acid color separation 5sec and 1% eosin stains 30sec through 1% haematoxylin immersion 15min, water logging respectively successively, rinses post-drying, resin mounting through water.

(6) fatty liver pathological grading builds high pathological diagnosis standard with reference to model.Under light microscopic, using "-", "+", " ++ " and " +++ " as four grades of fatty liver degeneration, with corresponding " normally ", " mild fatty liver ", " moderate fatty liver " and " severe fatty liver " clinically respectively.Normal or the accidental fat vacuole of hepatocyte structure is "-" level; 1/3 hepatic cell fattydegeneration in lobules of liver, involves lobules of liver quantity <50% for "+" level; 2/3 hepatic cell fattydegeneration in lobules of liver, involves lobules of liver quantity <50%, or 1/3 hepatic cell fattydegeneration in lobules of liver, involves lobules of liver quantity >50% for " ++ " level; >2/3 hepatic cell fattydegeneration in lobules of liver, involves lobules of liver quantity >50% for " +++ " level.

The fat stains of hepatic tissue frozen section and image quantitative analysis

(1) hepatic tissue cuts size and is about 1cm × lcm × lcm, fixes with 10% formaldehyde, is stored in-20 DEG C of refrigerators to make frozen section.

(2)-20 DEG C of descending frozen sections, slice thickness is 8 μm, to speckle with the slide bonding die of collodion.

(3) section is put into 70% ethanol and wash 1min, the rear S Ⅳ that drips carries out dyeing 1 ~ 2min, more slightly washes with 70% ethanol, and flowing water is washed again, after redye with haematoxylin, running water to returning indigo plant, neutral glycerine mounting.

(4) test sample book of Morphologic stereology, often organize from 60 routine specimen and select 5 examples at random, under mirror, optional 2 kiernan's space visuals field clap 5, do not repeat, and often organizing photo has the lobules of liver visual field 5 × 5 × 2=50.Often open and all amplify 40 times.

(5) with Mike Audi Motic Med.6.0 computer software measurement positive area, gray scale, average optical and integral optical density.

(6) SPSS16.0 is adopted to carry out the statistical computation of data.

Serum ALT, AST, GGT and ALP assay

Automatic clinical chemistry analyzer detects ALT, AST, GGT and ALP content

The mensuration of serum TG, TC, HDL-C and LDL-C

Rat is anaesthetized successfully through urethane, and ventral aorta is taken a blood sample, and gets whole blood and is about 7ml, get supernatant in-20 DEG C of preservations, measure serum TG, TC, HDL-C and LDL-C respectively after the centrifugal 10min of low speed 1500rpm.SPSS16.0 is adopted to carry out the statistical computation of data.

3) result

See Fig. 1-6.

Normal group liver surface color is generally dark red, bright, medium hardness, and tangent plane is granular sensation slightly, without greasy feeling.The most liver surface yellowish of model group, liver volume obviously increases, and peplos is nervous, and partial liver is milk yellow, and tangent plane is greasy oil droplet, without granular sensation.All the other respectively fall between by administration group color, quality.

Table 1 is group liver wet weights and liver index respectively

* significant difference (P < 0.01) is had for comparing with normal group group

Compared with normal group, model group body weight, liver wet weights and liver index all significantly raise (P < 0.05); Compared with model group, each medication group all reduces rat body weight, liver wet weights regulating liver-QI index, wherein has significant difference, close to normal group to liver index.

HE dyeing hepatic tissue (lobus sinister) pathological section result

Table 2 pathological section com-parison and analysis

Image quantitative analysis result

Table 3 positive area, gray scale, average optical, integral optical density fat stains image quantitative analysis between each group compares

* for comparing with model group, there is significant difference (P<0.05)

Table 3 result shows, in positive area, gray scale, average optical and integral optical density 4 indexs, pharmaceutical composition high dose group of the present invention is compared model group and is all significantly improved, improvement level is all better than DONGBAO GANTAI matched group, and along with the rising of pharmaceutical composition dosage of the present invention, the scope that hepatic tissue cell is got involved and degree all make moderate progress.

Serum ALT, AST, GGT and ALP assay

Table 4 is group rat blood serum ALT, AST, GGT, ALP comparision contents respectively

* significant difference (P < 0.05) is had for comparing with normal group group; * has significant difference (P < 0.05) compared with model group

Table 4 result shows, model group ALT, AST, GGT and ALP all significantly raise (P<0.05), illustrates that model group rats hepatocyte injury is more serious; Compared with model group, each medication group reduces ALT, AST, GGT and ALP level all to some extent, significant difference (P<0.05).

Serum TG, TC, HDL-C and LDL-C measurement result

Table 5 serum TC, TG, HDL-C, LDL-C com-parison and analysis

Table 5 result shows, model group TC, TG and LDL-C content all raise, and TC and LDL-C Be very effective (P<0.05); HDL-C content reduces, and does not have statistical significance (P>0.05).Compared with model group, each medication group reduces TC, TG and LDL-C content all in various degree, increases HDL-C content, and pharmaceutical composition effect of the present invention is better than DONGBAO GANTAI.

Above-mentioned experimental result shows, pharmaceutical composition of the present invention, hepatic cell fattydegeneration in high fat diet animal model and liver lactone class are assembled and improves significantly or reversing effect, and serum total cholesterol and low-density lipoprotein cholesterol content can be reduced, high density lipoprotein increasing cholesterol level, shows that pharmaceutical composition of the present invention has good therapeutical effect to non-alcoholic fatty liver disease.

The clinical trial of test example 2 Drug therapy non-alcoholic fatty liver disease of the present invention

1) case selection and grouping:

Select in November ,-2012 in January, 2009 Second People's Hospital of Fujian Province's outpatient service and be in hospital nonalcoholic fatty liver disease 80 example, all meet calendar year 2001 Chinese Medical Association's hepatology branch's fatty liver and alcoholic liver disease group formulate non-alcoholic fatty liver disease diagnostic criteria.Wherein, man 48 example, female 32 example; 20 ~ 60 years old age, average (50.0 ± 1.6) year.Patient is divided into treatment group and matched group at random by medical priority, often organizes 40 examples.Two groups of patients equal not statistically significant of comparing difference (the equal >0.05 of P) in sex, age, Clinical typing and complication etc., has comparability.

Exclusion standard: excessive drinking; History of virus hepatitis, cirrhosis patients in decompensation; Severe cardiac, brain, kidney diaseases, malignant tumor; Psychosis; Gestation and age of sucking; Obesity.

Therapeutic Method: treatment group gives pharmaceutical composition of the present invention (being prepared by embodiment 1), every day 3 times, each 1 bag, often bag is equivalent to crude drug consumption is 1.32g; It is oral that matched group gives DONGBAO GANTAI, each 4, and every day 3 times, the course for the treatment of is 3 months.

2) observation index and detection method

Serologic detection: take patient's fasting blood early morning, detects liver function, blood fat indices.

Imaging examination: row liver B ultrasonic before and after treatment, observes the Imageology of liver.

Curative effect determinate standard: 1. fully recover: blood biochemistry index and B ultrasonic Radiologic imaging all transfer to normally.2. take a turn for the better: blood biochemistry index is still higher than normal reference value, but the comparatively front reduction for the treatment of; B ultrasonic iconography still has abnormal change, but the comparatively front improvement for the treatment of.3. invalid: blood biochemistry index and B ultrasonic Imageology nothing compared with before treatment is improved or increases the weight of.

Wherein judge lesion degree according to B ultrasonic feature: 1. mild fatty liver: luminous point is fine and closely woven, near field Echoenhance, far field echo is slightly decayed, and blood vessel structure is clear.2. moderate fatty liver: luminous point is fine and closely woven, front court Echoenhance, obviously, blood vessel structure is unintelligible in far field decay.3. severe fatty liver: luminous point is fine and closely woven, front court echo significantly strengthens, and far field echo is significantly decayed, and blood vessel structure can not be recognized.

3) result

Changes of liver function (table 6):

Before and after table 6 liang group treatment, changes of liver function compares

Compare with before treatment, after matched group and treatment group patient treatment, alanine aminotransferase (ALT), aspartate transaminase (AST), alkali phosphatase (ALP) and C-glutamyl transferase (C-GGT) all have and decline in various degree, and difference has statistical significance (P<0.01).In the rear liver function for the treatment of group treatment, all comparatively matched group decline is more obvious for every serum enzyme index, and difference has statistical significance (P<0.05).

Blood Lipid (table 7):

Before and after table 7 liang group treatment, Blood Lipid compares

Compare with before treatment, after two groups of treatments, cholesterol (TC), triacylglycerol (TG) level obviously decline, HDL-C (HDL-C) level obviously raises, and difference has statistical significance (P<0.01).After treatment group treatment, comparatively matched group is remarkable for each target improvement situation, and difference has statistical significance (P<0.05).

Imaging examination (table 8):

Before and after table 8 liang group treatment, Imaging examination compares

After treatment group treatment, fatty liver elimination factor is significantly higher than matched group, and difference has statistical significance (P<0.05).

Comprehensive therapeutic effect (table 9):

Table 9 liang group comparitive study

Group	Number of cases	Recovery from illness	Take a turn for the better	Invalid	Effectively total
						Matched group	40	4（10.0）	20（50.0）	16（40.0）	24（60.0）
Treatment group	40	10（25.0）	24（60.0）	6（15.0）	34（85.0）

Treatment group cure rate and total effective rate are significantly higher than matched group, and difference has statistical significance (P<0.05).

Clinical experiment shows, pharmaceutical composition of the present invention can obviously reduce serum transaminase, cholesterol and triacylglycerol, high density lipoprotein increasing cholesterol levels, has good liver-protecting and blood fat-reducing effect, effectively can treat non-alcoholic fatty liver disease.

Claims (9)

1. treat a pharmaceutical composition for non-alcoholic fatty liver disease, it is characterized in that: it is the preparation be prepared from by the crude drug of following weight proportion:

Herba Artemisiae Scopariae 7-13 part, Fel Ursi powder 0.2-0.4 part, Rhizoma Atractylodis Macrocephalae 4.2-7.8 part, Rhizoma Alismatis 6-12 part, Fructus Crataegi 7-13 part, Radix Bupleuri 4.2-7.8 part, Radix Glycyrrhizae 4.2-7.8 part;

Its formulation preparation method comprises following operating procedure:

(1) according to consumption proportion weighting raw materials;

(2) Herba Artemisiae Scopariae, Rhizoma Alismatis, the Rhizoma Atractylodis Macrocephalae, Fructus Crataegi, Radix Bupleuri, Radix Glycyrrhizae is got, first extracting in water, ethanol is added again to alcohol content 60-90%v/v after extracting solution is concentrated, after precipitate with ethanol, get supernatant, reclaim ethanol, concentrate or after drying, add Fel Ursi powder and pharmaceutically conventional adjuvant or complementary composition and be prepared into preparation.

2. pharmaceutical composition according to claim 1, is characterized in that: it is the preparation be prepared from by the crude drug of following weight proportion:

Herba Artemisiae Scopariae 10 parts, Fel Ursi powder 0.3 part, the Rhizoma Atractylodis Macrocephalae 6 parts, Rhizoma Alismatis 9 parts, Fructus Crataegi 10 parts, Radix Bupleuri 6 parts, 6 parts, Radix Glycyrrhizae.

3. pharmaceutical composition according to claim 1 and 2, is characterized in that: described Radix Bupleuri is Radix Bupleuri bupleurum chinensedC..

4. the pharmaceutical composition according to claim 1-3 any one, is characterized in that: described preparation is oral formulations.

5. the preparation method of pharmaceutical composition described in claim 1-4 any one, is characterized in that: it comprises following operating procedure:

(1) according to consumption proportion weighting raw materials;

(2) Herba Artemisiae Scopariae, Rhizoma Alismatis, the Rhizoma Atractylodis Macrocephalae, Fructus Crataegi, Radix Bupleuri, Radix Glycyrrhizae is got, first extracting in water, ethanol is added again to alcohol content 60-90%v/v after extracting solution is concentrated, after precipitate with ethanol, get supernatant, reclaim ethanol, concentrate or after drying, add Fel Ursi powder and pharmaceutically conventional adjuvant or complementary composition and be prepared into preparation.

6. preparation method according to claim 5, is characterized in that: in step (2), alcohol content is 65-80%v/v.

7. the purposes of the pharmaceutical composition described in claim 1-4 any one in the medicine of preparation treatment non-alcoholic fatty liver disease.

8. purposes according to claim 7, is characterized in that: described medicine is the medicine for the treatment of Primary nonalcoholic fatty liver disease.

9. the pharmaceutical composition described in claim 1-4 any one is being prepared the liver protecting and ALT lowering or is being improved the purposes in the medicine of dyslipidemia.