CN103301414B - Gout, short kidney are arranged, repair kidney damages pharmaceutical composition and its preparation method and application - Google Patents
Gout, short kidney are arranged, repair kidney damages pharmaceutical composition and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of gout, short kidney row, repair kidney damage pharmaceutical composition and its preparation method and application, said composition is made up of the component of following weight portion: Rhizoma Polygoni Cuspidati 1-5 weight portion, Rhizoma Curcumae Longae 1-5 weight portion, Radix Salviae Miltiorrhizae 1-5 weight portion, Rhizoma Panacis Japonici 1-5 weight portion.This pharmaceutical composition prepares by alcohol extraction or water extraction method.Aforementioned pharmaceutical compositions has significance effect for gout, reparation injury of kidney, increase kidney discharge capacity.
Description
Technical field
The invention belongs to technical field of Chinese medicines, be specifically related to a kind of gout, short kidney row, the pharmaceutical composition repairing injury of kidney and its preparation method and application.
Background technology
Gout a kind ofly increases metabolic disease for feature with uric acid in blood.Along with our people's growth in the living standard, the sickness rate of hyperuricemia and gout rises year by year, persistence gout easily causes renal dysfunction, is the key factor of bringing out ischemic heart desease, renal calculus, obesity, hyperlipidemia, diabetes, hypertension etc.In the treatment of gout, there is toxic and side effects in various degree and limitation in Western medicine, as allopurinol class medicine, only can suppress the activity of xanthine oxidase, it does not have the effect promoting that uric acid urine is discharged, also do not have the effect of repairing metabolic arthritis injury of kidney, common adverse effect comprises: 1. allergic rash, urticaria, drug fever, hypereosinophilic etc.; 2. myelosuppressive leukopenia, hemolytic anemia; 3. toxic hepatitis or a glutamate pyruvate transaminase raise; 4. vasculitis and eye infringement; 5. xanthic calculus; As benzbromarone class medicine only can promote that uric acid is got rid of in urine, but can increase the weight of the damage of uric acid to kidney, major side effects has gastrointestinal dysfunction, renal colic, gout acute attack, erythra etc., accidental bone marrow depression.
Chinese medicine has the advantages such as medicine source is abundant, advantage of lower cost, determined curative effect and toxic and side effects are little, therefore, develops a kind of gout and repaiies pharmaceutical composition that kidney damages and the treatment being applied to gout has great importance.
Summary of the invention
The object of this invention is to provide a kind of can effectively gout, short kidney row, repair the pharmaceutical composition of injury of kidney.
Another object of the present invention is to provide the preparation method of aforementioned pharmaceutical compositions.
The object of the invention is to realize in the following manner:
A pharmaceutical composition for gout, short kidney row, reparation injury of kidney, this pharmaceutical composition is made up of the component of following weight portion: Rhizoma Polygoni Cuspidati 1-5 weight portion, Rhizoma Curcumae Longae 1-5 weight portion, Radix Salviae Miltiorrhizae 1-5 weight portion, Rhizoma Panacis Japonici 1-5 weight portion.
Aforementioned pharmaceutical compositions is preferably made up of the component of following weight portion:
Rhizoma Polygoni Cuspidati 1-3 weight portion, Rhizoma Curcumae Longae 1-3 weight portion, Radix Salviae Miltiorrhizae 1-3 weight portion, Rhizoma Panacis Japonici 1-3 weight portion.
The said composition further preferred component by following weight portion is made:
Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 2 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 1 weight portion;
Or Rhizoma Polygoni Cuspidati 2 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 1 weight portion;
Or Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 2 weight portion, Rhizoma Panacis Japonici 1 weight portion;
Or Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 2 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 2 weight portion;
Or Rhizoma Polygoni Cuspidati 2 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 2 weight portion;
Or Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 2 weight portion, Rhizoma Panacis Japonici 2 weight portion;
Or Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 2 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 3 weight portion;
Or Rhizoma Polygoni Cuspidati 2 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 3 weight portion;
Or Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 2 weight portion, Rhizoma Panacis Japonici 3 weight portion;
Or Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 1 weight portion;
Or Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 2 weight portion;
Or Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 3 weight portion;
Or Rhizoma Polygoni Cuspidati 2 weight portion, Rhizoma Curcumae Longae 2 weight portion, Radix Salviae Miltiorrhizae 2 weight portion, Rhizoma Panacis Japonici 1 weight portion;
Or Rhizoma Polygoni Cuspidati 2 weight portion, Rhizoma Curcumae Longae 2 weight portion, Radix Salviae Miltiorrhizae 2 weight portion, Rhizoma Panacis Japonici 3 weight portion;
Or Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 2 weight portion;
Or Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 1 weight portion.
Above-mentioned composition is most preferably made up of the component of following weight portion:
Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 1 weight portion.
Described compositions can make preparation with pharmaceutically acceptable carrier.Described preparation can be oral liquid, pill, tablet, powder, granule, capsule, drop pill or injection.
Above-mentioned composition prepares by alcohol extracting method, concrete preparation method comprises the following steps: by Rhizoma Polygoni Cuspidati, Rhizoma Curcumae Longae, Radix Salviae Miltiorrhizae, Rhizoma Panacis Japonici mass concentration is that the alcohol heating reflux of 10%-90% extracts 1-3 time, each extraction 1-3 hour, the amount of each ethanol is respectively 8-12 times of weight of medical material gross weight, filter, merging filtrate, concentrated.Preferred service property (quality) concentration is that the alcohol heating reflux of 60%-70% extracts.Can after first decompression recycling ethanol time concentrated, then heating be concentrated into concentration be 0.54g crude drug/ml or heating concentrated after to complement to concentration with distilled water be 0.54g crude drug/ml.
Above-mentioned composition also can be prepared by water extraction method, concrete preparation method comprises the following steps: take Rhizoma Polygoni Cuspidati, Rhizoma Curcumae Longae, Radix Salviae Miltiorrhizae, Rhizoma Panacis Japonici, by water heating extraction 1-3 time, each extraction 0.5-1 hour, the amount of each water is respectively medical material gross weight 8-12 times amount, filters, merging filtrate, concentrated.
Pharmaceutical composition of the present invention can prepared gout, repair injury of kidney, urge to apply in the medicine of kidney discharge capacity.The injury of kidney that described injury of kidney is mainly caused by hyperuricemia.
Rhizoma Polygoni Cuspidati, RhizomaPolygoniCuspidati is Polygonaceae Polygonaceae Polygonum Polygonum plant polygonum cuspidatum PolygonumcuspidatumSieb.etZucc., dry rhizome and root, effect: heat-clearing and toxic substances removing, promoting the function of the gallbladder to alleviate jaundice, expelling wind and removing dampness, dissipating blood stasis analgesic therapy, relieving cough and resolving phlegm.Rhizoma Curcumae Longae, RhizomaCurcumaeLongae, the dry rhizome of zingiberaceous plant Rhizoma Curcumae Longae CurcumalongaL., effect: removing blood stasis circulation of qi promoting, inducing menstruation to relieve menalgia; Radix Salviae Miltiorrhizae Salviamiltiorrhiza is the rhizome of salvia Salviamiltiorrhiza, effect: promoting blood flow to regulate menstruation, and stasis-dispelling and pain-killing, removing heat from blood eliminating carbuncle, clear away heart-fire relieving restlessness, nourishing blood to tranquillize the mind.Rhizoma Panacis Japonici, RhizomaPanacisJaponici, the dry rhizome of Araliaceae Rhizoma Panacis Japonici PanaxjaponicusC.A.Mey., effect: strengthening by means of tonics, eliminating stasis to stop pain, hemostasis is eliminated the phlegm.The advantage of the comprehensive above-mentioned raw materials medical material of inventor, in conjunction with modern new drug research means, develop can effectively gout, short kidney row, repair the pharmaceutical composition of injury of kidney, this pharmaceutical composition also has use safety, rapid-action feature.
Beneficial effect of the present invention compared with the prior art: prescription of the present invention has pole significance effect for reduction animal pattern serum uric acid, for promotion uraturia discharge capacity, there is pole significance effect, for inhibition animal livers xanthine oxidase, there is pole significance effect.Therefore, prescription of the present invention has the pole significance effect of repairing the injury of kidney that hyperuricemia causes.Prescription of the present invention reduces the main path of animal pattern serum uric acid effect by repairing injury of kidney, promoting kidney discharge capacity and realize.
Below by way of concrete test example, above-mentioned effect is further described:
Drug regimen is made up of Rhizoma Polygoni Cuspidati, Rhizoma Curcumae Longae, Radix Salviae Miltiorrhizae, Rhizoma Panacis Japonici 4 herbal medicine.Now list for uric acid resisting experiment and repair the drug regimen sequence number of metabolic arthritis injury of kidney experiment and the part by weight relation of the combination of 4 herbal medicines under this sequence number.
Prescription 1 group: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=1:2:3:1
Prescription 2 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=2:3:1:1
Prescription 3 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=3:1:2:1
Prescription 4 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=1:2:3:2
Prescription 5 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=2:3:1:2
Prescription 6 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=3:1:2:2
Prescription 7 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=1:2:3:3
Prescription 8 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=2:3:1:3
Prescription 9 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=3:1:2:3
Prescription 10 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=1:1:1:1
Prescription 11 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=1:1:1:2
Prescription 12 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=1:1:1:3
Prescription 13 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=2:2:2:1
Prescription 14 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=2:2:2:3
Prescription 15 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=3:3:3:2
Prescription 16 groups: Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: Rhizoma Panacis Japonici=3:3:3:1
Test example 1: adopt above-mentioned 17 groups of medicines to carry out gout experiment:
1. experiment material
1.1 instruments: ISO9001 type electronic analytical balance (Beijing Sai Duolisi instrument system company limited); TDL80-2B type low speed centrifuge (Anting Scientific Instrument Factory, Shanghai); RT-6000 type enzyme-linked immunosorbent assay instrument (Shenzhen Lei Du Life Science limited company); Metabolic cage; Refiner; Liquid-transfering gun; Claim Mus scale; Balance.
1.2 reagent and reagent: testing uric acid test kit (lot number: 20120511 is purchased from Nanjing and builds up Bioengineering Research Institute); Xanthine oxidase measures test kit (lot number: 20120512 is purchased from Nanjing and builds up Bioengineering Research Institute); Coomassie brilliant blue protein determination kit (lot number: 20120427 is purchased from Nanjing and builds up Bioengineering Research Institute); TONGFENGDING capsule (Zhonghui Pharmacy Co., Ltd., Chengdu's lot number: 110507); (bio tech ltd is built up in Nanjing to Oteracil Potassium, lot number: 120204), xanthine (sigma, lot number: 120201), Rhizoma Polygoni Cuspidati, Radix Salviae Miltiorrhizae, Rhizoma Curcumae Longae, Rhizoma Panacis Japonici (all purchased from Nanjing first sign pharmacy).
1.3 animals: Kunming mouse, male, body weight 18-22g, 210, cleaning grade (animal field, green hill, Nanjing provides).
2. the preparation of various medicinal liquid
The preparation of prescription 16 alcohol extraction medicinal liquid: take Rhizoma Polygoni Cuspidati 8.1g according to No. 16 prescription ratio, Rhizoma Curcumae Longae 8.1g, Radix Salviae Miltiorrhizae 8.1g, Rhizoma Panacis Japonici 2.7g, medicine total amount is 27g, extract three times with the alcohol heating reflux that concentration is 60%, extract 2,2,1.5 hours respectively, the amount of each ethanol is respectively 12,10,8 times of weight of medical material.Filter, merge three filtrates, decompression recycling ethanol is to 50ml, 50ml is complemented to distilled water less than during 50ml, be settled to 100mL with the CMC-Na solution that concentration is 0.5% again, obtain No. 16 prescription alcohol extraction medicinal liquid that concentration is 0.27g/ml, the pharmacological evaluation of pending gout.
The configuration of prescription 1-15 group alcohol extraction medicinal liquid: take Rhizoma Polygoni Cuspidati, Rhizoma Curcumae Longae, Radix Salviae Miltiorrhizae, Rhizoma Panacis Japonici respectively according to 1-15 prescription ratio, these 15 groups of medical material weight reach 27g respectively, preparation method is with the preparation method of No. 16 prescription alcohol extraction medicinal liquid, be mixed with the prescription 1-15 alcohol extraction medicinal liquid that concentration is 0.27g/ml, the pharmacological evaluation of pending gout.
The preparation of prescription 16 water extraction medicinal liquid: take Rhizoma Polygoni Cuspidati 8.1g, Rhizoma Curcumae Longae 8.1g, Radix Salviae Miltiorrhizae 8.1g according to No. 16 prescription ratio, Rhizoma Panacis Japonici 2.7g, medicine total amount is 27g, by water heating extraction three times, extract 1,1,0.5 hour respectively, the amount of each water is respectively 12,10,8 times amount of medical material.Filter, merge three filtrates, water-bath is concentrated into 50ml, 50ml is complemented to distilled water less than during 50ml, be settled to 100mL with the CMC-Na solution that concentration is 0.5% again, obtain the water extraction medicinal liquid that concentration is No. 16 prescription of 0.27g/ml, the pharmacological evaluation of pending gout.
The configuration of modeling agent: precision takes Oteracil Potassium 0.6015g, xanthine 0.3072g pours in 100ml cillin bottle, measures 0.5%CMC-Na solution 60ml, ultrasonic 50min, dissolved substance, obtains containing 10mg/ml Oteracil Potassium and the xanthic modeling agent of 5mg/ml.Put in 2-8 DEG C of refrigerator and preserve.Administration volume is 20ml/kg, Oteracil Potassium dosage 200mg/kg, xanthine dosage 100mg/kg.
The configuration of positive drug TONGFENGDING capsule solution: computational methods prepared by TONGFENGDING capsule liquid: TONGFENGDING capsule adult dosage be one time 4, three times on the one, every 0.4g, i.e. 4.8g.Carry out the conversion of dosage according to people and Mouse Weight scaling method, mice dosage is 0.72g/kg.
The body weight (60 ㎏) (W is coefficient of converting: people and mice administration conversion coefficient are 9.01) of dosage (g/kg)=W × adult's dosage every day (the g)/adult of mice.
The configuration of TONGFENGDING capsule solution: precision takes 1.8056g TONGFENGDING powder in conical flask, adds the CMC-Na solution of 50mL0.5%, ultrasonic dissolution, obtaining liquor strength is 36.0mg/mL, and mice administration volume is 0.2mL/10g, and dosage is 0.72g/kg.
3. mice uric acid resisting experimental technique and result
Be the Male Kunming strain mice of 18 ~ 22g by 210 body weight, be divided into 21 groups at random, often organize 10.Be respectively blank group, model group, allopurinol group, TONGFENGDING group, 17 groups of medicament composing prescription groups of the present invention.Gavage, mice administration volume is 20mL/kg, and metabolic arthritis model group gives the CMC-Na solution of 0.5%, other each compound recipe gives corresponding medicinal liquid, and continue gavage 6d, dosage is respectively allopurinol group 0.72g/kg, TONGFENGDING capsule 0.72g/kg, medicament composing prescription 5.4g/kg of the present invention.Modeling in 7th day, administration volume is 20mL/kg, and the isopyknic normal saline of blank group lumbar injection, all the other respectively organize equal lumbar injection modeling agent.After modeling, 1h gives each group of mice relative medicine again, and the mode of administration, administration volume, dosage are the same.Carry out eyeground vein clump to each group of mice after last administration 1h and get blood, amount for taking blood is about 0.5ml, and de-neck puts to death mice, dissects mice simultaneously and gets appropriate liver organization.The urine of 1h, merges urine after collecting mice modeling and after administration.
The process of sample:
Blood sample treatments: modeling, after 2 hours, is got blood from mice eyeground vein clump and is about 0.5mL, by the blood sample of each group of mice, room temperature places 1h, and after blood sample coagulation, centrifugal (4000rpm/min, 10min), is placed in-20 DEG C of Refrigerator stores by being separated the serum obtained, for subsequent use.
Liver process: precision takes each group of murine liver tissue and is about 0.05g, add the normal saline of 9 times amount, be placed in, in 2mL centrifuge tube, homogenate (10000rpm, 1min), by centrifugal for each homogenate (4000rpm/min, 10min), Aspirate supernatant, the activity (XOD) of hepatic tissue Tot Prot to be measured and xanthine oxidase thereof.
Urine process: keep the urine collected in mind urine volume, 50 DEG C of heating, after 5min, dilute with water 10 times, obtains urine sample to be measured immediately.
Sample determination method measures the vigor of xanthine oxidase (XOD) of serum uric acid value, urine uric acid level, liver organization liquid respectively by kit method.
Experimental result the data obtained adopts EXCEL2003 software, with
represent.Adopt significance test between one factor analysis of variance group, between two groups, sample compares employing t inspection, has statistical significance with P ﹤ 0.05.The results are shown in Table 1, table 2 and table 3.
Table 1 respectively group medicinal liquid affects experimental result to mice serum uric acid content
Note:
Δcompare with blank group,
Δp ﹤ 0.05,
Δ Δp ﹤ 0.01;
*compare with model group,
*p ﹤ 0.05,
*p ﹤ 0.01.
Experimental result shows: model group compare with blank group have extremely significant difference (
Δ Δp ﹤ 0.01) illustrative experiment modeling success; TONGFENGDING group and compare with model group (
*p ﹤ 0.05), illustrate that TONGFENGDING has the effect of the reduction serum uric acid of significance; Prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group, allopurinol group compare with model group (
*p ﹤ 0.01), illustrate that prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group, allopurinol group have the effect reducing serum uric acid extremely significantly.
Table 2 respectively organizes medicinal liquid to mouse liver XOD activity influence result
Note:
Δcompare with blank group,
Δp ﹤ 0.05,
Δ Δp ﹤ 0.01;
*compare with model group,
*p ﹤ 0.05,
*p ﹤ 0.01.
Experimental result shows: model group compares P ﹤ 0.05 with blank group, shows modeling success; TONGFENGDING group and comparing with model group have significant difference (
*p ﹤ 0.05), illustrate that positive drug TONGFENGDING has the significance effect of certain suppression hepatic tissue XOD activity; Prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group, allopurinol group compare with model group (
*p ﹤ 0.01), illustrate that prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group, allopurinol group have and suppress XOD active extremely significantly.
Table 3 respectively group medicinal liquid affects result to mice urine urate excretion amount
Note:
*compare with model group,
*p ﹤ 0.05,
*p ﹤ 0.01.
Experimental result shows: TONGFENGDING group, allopurinol group and do not have significant difference with model group, illustrates that positive drug TONGFENGDING, allopurinol do not have the active function promoting uraturia excretion; Prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group compare with model group (
*p ﹤ 0.01), prescription 1-16 alcohol extract is described, prescription 16 Aqueous extracts have extremely significantly promote uraturia excretion active function.
The anti-mouse metabolic arthritis injury of kidney experiment of the above-mentioned 17 groups of medicines of test example 2.
1. experiment material
1.1 medicines and reagent: TONGFENGDING capsule (Chengdu Zhong Jiang pharmaceutical Co. Ltd, lot number: 090104), ebutol (Hangzhou Minsheng Pharmaceutical Group Co, lot number: T09L142), adenine (Zhejiang Rui Xin Pharmaceutical limited company, lot number: 091105), (bio tech ltd is built up in Nanjing to testing uric acid test kit, lot number: 20090120), (bio tech ltd is built up in Nanjing to creatinine assay test kit, lot number: 20090106), (bio tech ltd is built up in Nanjing to blood urea nitrogen testing cassete, lot number: 20090113), ethanol, chloroform, petroleum ether etc. (are CR, Nanjing chemical reagent factory).Rhizoma Polygoni Cuspidati, Radix Salviae Miltiorrhizae, Rhizoma Curcumae Longae, Rhizoma Panacis Japonici (all purchased from Nanjing first sign pharmacy).
1.2 laboratory animals: Kunming mouse, male, body weight 18-22g, 210, cleaning grade (animal field, green hill, Nanjing provides).
1.3 experiment equipments: B-260 type thermostat water bath (Shanghai Yarong Biochemical Instrument Plant); T80-2B type low speed centrifuge (Anting Scientific Instrument Factory, Shanghai); DG5033A type microplate reader (Shanghai Kun Ken biochemical industry company limited); Electronic balance, refiner, tweezers, dissecting scissors, centrifuge tube, liquid-transfering gun.
2. the various preparation for reagent liquid
The preparation of prescription 16 alcohol extraction medicinal liquid: take Rhizoma Polygoni Cuspidati 8.1g according to No. 16 prescription ratio, Rhizoma Curcumae Longae 8.1g, Radix Salviae Miltiorrhizae 8.1g, Rhizoma Panacis Japonici 2.7g, medicine total amount is 27g, extract three times with the alcohol heating reflux that concentration is 60%, extract 2,2,1.5 hours respectively, the amount of each ethanol is respectively 12,10,8 times of weight of medical material.Filter, merge three filtrates, decompression recycling ethanol, to 50ml, complements to 50ml less than during 50ml with distilled water, then is settled to 100mL with the CMC-Na solution that concentration is 0.5%, obtains No. 16 prescription alcohol extraction medicinal liquid that concentration is 0.27g/ml.
The configuration of prescription 1-15 group alcohol extraction medicinal liquid: take Rhizoma Polygoni Cuspidati, Rhizoma Curcumae Longae, Radix Salviae Miltiorrhizae, Rhizoma Panacis Japonici respectively according to 1-15 prescription ratio, these 15 groups of medical material weight reach 27g respectively, preparation method, with the preparation method of No. 16 prescription alcohol extraction medicinal liquid, is mixed with the prescription 1-15 alcohol extraction medicinal liquid that concentration is 0.27g/ml.
The preparation of prescription 16 water extraction medicinal liquid: take Rhizoma Polygoni Cuspidati 8.1g, Rhizoma Curcumae Longae 8.1g, Radix Salviae Miltiorrhizae 8.1g according to No. 16 prescription ratio, Rhizoma Panacis Japonici 2.7g, medicine total amount is 27g, by water heating extraction three times, extract 1,1,0.5 hour respectively, the amount of each water is respectively 12,10,8 times amount of medical material.Filter, merge three filtrates, water-bath is concentrated into 50ml, complements to 50ml less than during 50ml with distilled water, then is settled to 100mL with the CMC-Na solution that concentration is 0.5%, obtains the water extraction medicinal liquid that concentration is No. 16 prescription of 0.27g/ml.
The configuration of positive drug TONGFENGDING capsule solution: computational methods prepared by TONGFENGDING capsule liquid: TONGFENGDING capsule adult dosage be one time 4, three times on the one, every 0.4g, i.e. 4.8g.Carry out the conversion of dosage according to people and Mouse Weight scaling method, mice dosage is 0.72g/kg.
The body weight (60 ㎏) (W is coefficient of converting: people and mice administration conversion coefficient are 9.01) of dosage (g/kg)=W × adult's dosage every day (the g)/adult of mice.
The configuration of TONGFENGDING capsule solution: precision takes 1.8056g TONGFENGDING powder in conical flask, adds the CMC-Na solution of 50mL0.5%, ultrasonic dissolution, obtaining liquor strength is 36.0mg/mL, and mice administration volume is 0.2mL/10g, and dosage is 0.72g/kg.
The CMC-Na solution preparation of 0.5%: take CMC-Na2.5 gram and add in 500mL pure water to heat and ultrasonicly make it dissolve, hold over night.
The preparation of normal saline: take 4.5041 grams of sodium chloride, is dissolved in 500mL pure water and get final product.
The preparation of modeling agent adenine and ebutol mixed solution: take adenine 1.040 grams and ebutol 1.500 grams, by its ultrasonic dissolution in the CMC-Na solution of 0.5% of 60mL and get final product, the concentration of adenine, ebutol is respectively 17.33mg/mL, 25mg/mL.The gavage volume 0.2mL/10g of modeling.Modeling agent adenine dosage is 346mg/kg, and ebutol dosage is 500mg/kg.
3 experimentations
Get 210 Male Kunming strain mice, body weight is 20-22g, is divided into 21 groups at random, often organizes each 10.Be respectively blank group, model group, positive control drug TONGFENGDING group, positive control drug allopurinol group, medicament composing prescription of the present invention totally 17 groups.Be known as by each group of mouse heavily, every morning 8:30 beginning outer the filling respectively each group of mice of blank group gives modeling agent.Modeling method adopts mouse stomach adenine and ebutol modeling agent solution to set up metabolic arthritis type renal injury model, the volume of modeling gavage is 0.2mL/kg, modeling dosage is: adenine 346mg/kg(concentration is 17.33mg/mL), ebutol 500mg/kg(concentration is 25mg/mL).Continuous modeling 21 days, modeling and administration are carried out simultaneously.Evening, 20:30 started to fill with each group of mice to give corresponding medicine, gavage volume is 0.2mL/10g, blank group and model group gavage the CMC-Na solution of 0.5%, the dosage of each group is respectively: dosage is respectively allopurinol group 0.72g/kg, TONGFENGDING capsule 0.72g/kg, medicament composing prescription of the present invention is 5.4g/kg respectively.Claim its weight every 7 days to each group of mice, redefine the modeling of each group and the gavage volume of administration.Eye socket was carried out to each group of mice in the 7th day, the 21st day and gets blood in experiment, (4000rp/ is per minute for separation of serum, centrifugal 15 minutes), be placed in-20 DEG C of Refrigerator stores, test serum blood urea nitrogen (BUN), serum creatinine (SCr) 2 indexs by being separated the serum obtained.The experimental result of comparative measurements index, thus each administration group of analysis and inspection is to the repair of injury of kidney.
The modeling mechanism of gouty injury of kidney: high concentration adenine generates pole by the effect of xanthine oxidase and is insoluble in 2 of water; 8-dihydroxy adenine; 2; 8-dihydroxy adenine is deposited on renal tubules; have impact on the excretion of nitrogen matter compound, cause azotemia, toxin to be accumulated and electrolyte and disorder of amino acid metabolism, ebutol suppresses renal tubules to uric acid excretion; make underexcretion and cause hyperuricemia, thus causing chronic nephritis and the renal failure of metabolic arthritis.
4 each experimental index measure
The mensuration of creatinine content and urea nitrogen content is carried out according to the assay method of creatinine reagent box, blood urea nitrogen test kit.
Computing formula:
Serum creatinine (μm ol/L)=concentration of standard solution × 11
*× (A measures pipe-A blank tube)/(A standard pipe-A blank tube).Wherein concentration of standard solution 10 μm of ol/L.
Urea nitrogen content (mmol/L)=concentration of standard solution × (A measures pipe-A blank tube)/(A standard pipe-A blank tube).Concentration of standard solution 10mmol/L.
5 experimental results
The data obtained SPSS11.5 statistical analysis software carries out statistical analysis, experimental result with
represent, refer to table 4,5.
Table 4: the experimental data of each group medicine mice serum blood urea nitrogen and analysis (mmol/L) (n=10)
Note: compare with blank group,
*p ﹤ 0.05,
*p ﹤ 0.01; Compare with model group,
▲p ﹤ 0.05,
▲ ▲p ﹤ 0.01.
Table 5: the experimental data of each group medicine mice serum creatinine and analysis (μm ol/L) (n=10)
Note: compare with blank group,
*p ﹤ 0.05,
*p ﹤ 0.01; Compare with model group,
▲p ﹤ 0.05,
▲ ▲p ﹤ 0.01.
6 discuss and sum up
(1) model group compares with blank group, in modeling after 7 days, the serum urea nitrogen of modeling after 21 days, serum creatinine value two indices all have pole significant difference (P ﹤ 0.01), shows the renal injury model modeling success that the mice persistence hyperuricemia that gavage adenine and ebutol cause causes.
(2) by serum analysis blood urea nitrogen index, in modeling after the 7th day, after the 21st day, prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group more all have pole significant difference (P ﹤ 0.01) with model group; And positive control drug allopurinol group, TONGFENGDING group compare with model group, all not there is the difference of pole significance.Experiment shows that the reparation injury of kidney effect in the renal injury model that prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group cause in hyperuricemia is obviously better than positive control drug allopurinol, TONGFENGDING.
(3) by serum analysis creatinine index, in modeling after the 7th day, after the 21st day, prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group compare with model group, all have pole significant difference (P ﹤ 0.01); And positive control drug allopurinol group, TONGFENGDING group compare with model group, all not there is the difference of pole significance.Experiment shows that the reparation injury of kidney effect in the renal injury model that prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group cause in hyperuricemia is obviously better than positive control drug allopurinol, TONGFENGDING.
Therefore, the injury of kidney effect that the reparation hyperuricemia that prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group have pole significance causes.Its effect is better than positive control drug allopurinol, TONGFENGDING significantly.
7 conclusions
Prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group have the pole significance effect reducing animal pattern serum uric acid, have the pole significance effect promoting uraturia discharge capacity, have the pole significance effect of inhibition animal livers xanthine oxidase.Prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group have the pole significance effect of repairing the injury of kidney that hyperuricemia causes.Prescription 1-16 alcohol extract group, prescription 16 Aqueous extracts group reduce the main path of animal pattern serum uric acid effect by repairing injury of kidney, increasing kidney discharge capacity and realize.Prescription 16 is namely according to weight ratio Rhizoma Polygoni Cuspidati: Rhizoma Curcumae Longae: Radix Salviae Miltiorrhizae: the drug effect of Rhizoma Panacis Japonici=3:3:3:1 is best.
Detailed description of the invention
The present invention is further illustrated below by way of specific embodiment.But the detail of embodiment only for explaining the present invention, should not be construed as limited overall technical solution.
Embodiment 1
Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 2 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 1 weight portion.
Preparation method: take Rhizoma Polygoni Cuspidati, Rhizoma Curcumae Longae, Radix Salviae Miltiorrhizae, the alcohol heating reflux of Rhizoma Panacis Japonici concentration 60% extracts three times, each extraction 2,2,1.5 hours, the amount of each ethanol is respectively 12,10,8 times of weight of medical material, filters, merge 3 filtrates, decompression recycling ethanol, concentrated.
Embodiment 2
Rhizoma Polygoni Cuspidati 2 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 1 weight portion.Preparation method is with embodiment 1.
Embodiment 3
Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 2 weight portion, Rhizoma Panacis Japonici 1 weight portion.Preparation method is with embodiment 1.
Embodiment 4
Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 2 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 2 weight portion.Preparation method is with embodiment 1.
Embodiment 5
Rhizoma Polygoni Cuspidati 2 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 2 weight portion.Preparation method is with embodiment 1.
Embodiment 6
Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 2 weight portion, Rhizoma Panacis Japonici 2 weight portion.Preparation method is with embodiment 1.
Embodiment 7
Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 2 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 3 weight portion.Preparation method is with embodiment 1.
Embodiment 8
Rhizoma Polygoni Cuspidati 2 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 3 weight portion.Preparation method is with embodiment 1.
Embodiment 9
Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 2 weight portion, Rhizoma Panacis Japonici 3 weight portion.Preparation method is with embodiment 1.
Embodiment 10
Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 1 weight portion.Preparation method is with embodiment 1.
Embodiment 11
Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 2 weight portion.Preparation method is with embodiment 1.
Embodiment 12
Rhizoma Polygoni Cuspidati 1 weight portion, Rhizoma Curcumae Longae 1 weight portion, Radix Salviae Miltiorrhizae 1 weight portion, Rhizoma Panacis Japonici 3 weight portion.Preparation method is with embodiment 1.
Embodiment 13
Rhizoma Polygoni Cuspidati 2 weight portion, Rhizoma Curcumae Longae 2 weight portion, Radix Salviae Miltiorrhizae 2 weight portion, Rhizoma Panacis Japonici 1 weight portion.Preparation method is with embodiment 1.The medicinal liquid obtained adds the acceptable adjuvant of conventional pharmaceutical and conventionally makes capsule.
Embodiment 14
Rhizoma Polygoni Cuspidati 2 weight portion, Rhizoma Curcumae Longae 2 weight portion, Radix Salviae Miltiorrhizae 2 weight portion, Rhizoma Panacis Japonici 3 weight portion.Preparation method is with embodiment 1.The medicinal liquid obtained adds the acceptable adjuvant of conventional pharmaceutical and conventionally makes granule.
Embodiment 15
Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 2 weight portion.Preparation method is with embodiment 1.The medicinal liquid obtained adds the acceptable adjuvant of conventional pharmaceutical and is pressed into tablet.
Embodiment 16
Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 1 weight portion.Preparation method is with embodiment 1.The medicinal liquid obtained adds customary adjuvant and conventionally makes pill.
Embodiment 17
Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 1 weight portion.
Preparation method: take Rhizoma Polygoni Cuspidati according to aforementioned proportion, Rhizoma Curcumae Longae, Radix Salviae Miltiorrhizae, Rhizoma Panacis Japonici, by water heating extraction three times, extract 1,1,0.5 hour, the amount of each water is respectively 12,10,8 times of weight of medical material at every turn.Filter, merge 3 filtrates, concentrated.
Claims (7)
1. a pharmaceutical composition for gout, short kidney row, reparation injury of kidney, is characterized in that said composition is made up of the component of following weight portion:
Get Rhizoma Polygoni Cuspidati 3 weight portion, Rhizoma Curcumae Longae 3 weight portion, Radix Salviae Miltiorrhizae 3 weight portion, Rhizoma Panacis Japonici 1 weight portion;
By Rhizoma Polygoni Cuspidati, Rhizoma Curcumae Longae, Radix Salviae Miltiorrhizae, Rhizoma Panacis Japonici concentration is that the alcohol heating reflux of 10%-90% extracts 1-3 time, each extraction 1-3 hour, and the amount of each ethanol is respectively 8-12 times of weight of medical material, filters, merging filtrate, concentrated;
Or take Rhizoma Polygoni Cuspidati, Rhizoma Curcumae Longae, Radix Salviae Miltiorrhizae, Rhizoma Panacis Japonici, by water heating extraction 1-3 time, extract 0.5-1 hour, the amount of each water is respectively 8-12 times of weight of medical material gross weight, filtration, and merging filtrate concentrates at every turn.
2. pharmaceutical composition according to claim 1, is characterized in that described compositions and pharmaceutically acceptable carrier make preparation.
3. pharmaceutical composition according to claim 2, is characterized in that described preparation is oral liquid, pill, tablet, powder, granule, capsule, drop pill or injection.
4. the preparation method of a pharmaceutical composition according to claim 1, it is characterized in that the method comprises the following steps: by Rhizoma Polygoni Cuspidati, Rhizoma Curcumae Longae, Radix Salviae Miltiorrhizae, Rhizoma Panacis Japonici concentration is that the alcohol heating reflux of 10%-90% extracts 1-3 time, each extraction 1-3 hour, the amount of each ethanol is respectively 8-12 times of weight of medical material, filter, merging filtrate, concentrated.
5. the preparation method of a pharmaceutical composition according to claim 1, it is characterized in that the method comprises the following steps: take Rhizoma Polygoni Cuspidati, Rhizoma Curcumae Longae, Radix Salviae Miltiorrhizae, Rhizoma Panacis Japonici, by water heating extraction 1-3 time, each extraction 0.5-1 hour, the amount of each water is respectively 8-12 times of weight of medical material gross weight, filters, merging filtrate, concentrated.
6. the application of pharmaceutical composition according to claim 1 in the medicine preparing gout, reparation injury of kidney, short kidney discharge capacity.
7. application according to claim 6, is characterized in that described injury of kidney is the injury of kidney caused by hyperuricemia.
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| 丹参降尿酸作用初步实验研究;刘艳等;《海峡药学》;20130115;第25卷(第1期);27-29 * |
| 姜黄降尿酸作用的实验研究;殷华峰等;《药学与临床研究》;20110430;第19卷(第2期);134-135 * |
| 虎杖提取物对实验性痛风的作用及机理分析;林金朝;《亚太传统医药》;20121130;第8卷(第11期);21-22 * |
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