CN101670077A - Attenuated compatibility formula of minor Chinese thorowax root decoction - Google Patents

Abstract

The invention provides a novel attenuated compatibility formula of a minor Chinese thorowax root decoction. The formula reduces the proportion of a hepatotoxicity medicinal material scutellaria root,increases the proportion of pinellia tuber, and comprises the following components in portion by weight: 2.143 portions of Chinese thorowax root, 1.107 portions of baical skullcap root, 1.107 portionsof ginseng, 2.107 portions of the pinellia tuber, 1.107 portions of ginger, 0.893 portion of liquorice root, and 1.536 portions of Chinese date. Compared with the minor Chinese thorowax root decoction with the original formula in 'Treatise on Cold Pathogenic Diseases' written by Zhang Zhongjing, the remixed minor Chinese thorowax root decoction greatly attenuates the hepatotoxicity and simultaneously keeps the original curative effect of the minor Chinese thorowax root decoction.

Description

Attenuated compatibility formula of minor Chinese thorowax root decoction

Technical field

The present invention relates to the field of Chinese medicines, relate more specifically to a kind of new attenuated compatibility formula of minor Chinese thorowax root decoction.

Background technology

Herba Sidae Rhombifoliae soup comes from treatise on Febrile Diseases, form by Radix Bupleuri, Radix Scutellariae, Radix Ginseng, the Rhizoma Pinelliae, Radix Glycyrrhizae Preparata, Rhizoma Zingiberis Recens, Fructus Jujubae seven flavor medicine, it is the representative prescription for the treatment of SHAO YANG disease by mediation, on clinical department of internal medicine, use very extensive, Herba Sidae Rhombifoliae soup plus-minus treatment digestive system commonly used, blood circulation, respiratory system, urinary system, nervous system disease and psychosis ^[1]

In recent years the toxin for liver side effect of relevant Herba Sidae Rhombifoliae soup receives increasing concern.External Itoh etc. [Chen Damin. the hepatic injury that the medical herbs Herba Sidae Rhombifoliae soup causes. foreign medical science digestive system disease fascicle 1996; 16 (1): 61-65] report 40 examples are taken the Herba Sidae Rhombifoliae soup patient, there is 9 examples transaminase in the process of taking to raise and jaundice occurs, the hepatic tissue biopsy turns out to be acute liver injury, liver function recovery is normal after the drug withdrawal, hepatic injury appearred again after wherein 4 examples were taken medicine once more, show that Herba Sidae Rhombifoliae soup can bring out acute hepatitis, its mechanism may be relevant with the toxicity of the crude drug of Radix Bupleuri or Radix Scutellariae.It is reported [the gold boat. Japanese Chinese prescription medicine preparation side effect is analyzed. foreign medical science Chinese medicine fascicle 1997; 19 (3): 62-66] [Gan Dianhui, Deng the lady in the moon. the part Chinese herbal medicine causes the report summary [J] of medicamentous liver lesion. the Guangxi Chinese medicine, 2004,6:5-7], Herba Sidae Rhombifoliae soup is widely used in the chronic hepatopathy patient for a long time as the medicine of anti-liver injury, and the result causes acute hepatitis after average 2 months incubation period, the fat-like pathological changes and the fibrosis of the property necrosis of Microscopic examination showed lobules of liver central authorities, microcapsule blister, the performance [3,4] of cholestasis is arranged in two biopsy hepatic tissues.Also there is the scholar to report [Gu Wenjun, Liu Chunhong, Shen Xizhong. the liver toxicity of medical herbs [J]. world's clinical medicine, 2003,24:217-226.] untoward reaction of Herba Sidae Rhombifoliae soup and similar side's extractant thereof, wherein many cases are liver function injury (Herba Sidae Rhombifoliae soup 5 examples, little Radix Bupleuri are closed bavin Piao soup, little Radix Bupleuri adds each 1 example of Radix Platycodonis Shigao Tang).

In view of the research to each single medical material in the Herba Sidae Rhombifoliae soup at present focuses mostly on aspect chemical composition analysis and the pharmacological research,, do not see the research of its intoxicating composition and mechanism of toxication to the rarely seen report of the toxicological study of each single medicine of distinguishing the flavor of.In view of the toxin for liver side effect of Herba Sidae Rhombifoliae soup appearance clinically at present, therefore, the Herba Sidae Rhombifoliae soup composition of prescription of new attenuation need be developed in this area simultaneously, is guaranteeing to reduce its hepatotoxicity greatly on the former powerful basis of Herba Sidae Rhombifoliae soup.

Summary of the invention

The present invention confirms the liver toxicity of Herba Sidae Rhombifoliae soup prescription, and clear and definite causes hepatotoxic medical material in this prescription.

Purpose of the present invention just provides a kind of new attenuated compatibility formula of minor Chinese thorowax root decoction.

In the present invention, provide a kind of Herba Sidae Rhombifoliae soup composition of prescription, medical material and part by weight thereof that this prescription comprises are as follows,

2.143 parts of Radix Bupleuri, 1.107 parts of Radix Scutellariaes, 1.107 parts of Radix Ginsengs, 2.107 parts of the Rhizoma Pinelliaes, 1.107 parts in Rhizoma Zingiberis Recens, 0.893 part in Radix Glycyrrhizae, 1.536 parts in Fructus Jujubae

In another preference, described prescription is to the perform toxic attenuation of liver.

Description of drawings

Fig. 1 mtt assay detects different method for making Herba Sidae Rhombifoliae soup to the active influence of human hepatoma cell strain

Fig. 2 mtt assay detects the different compatibilities side of subtracting decocting liquid to the active influence of human hepatoma cell strain

The specific embodiment

The human liver cell toxicity vitro detection of embodiment 1. different composition of prescription Herba Sidae Rhombifoliae soup

1. material and instrument

1.1 medicine and reagent Radix Bupleuri, Rhizoma Pinelliae Preparata, Radix Glycyrrhizae Preparata, Fructus Jujubae are available from Shanghai Kangqiao Medicinal Materials Electuary Co., Ltd.; Radix Scutellariae is available from Shanghai Dehua National Medicine Products Co., Ltd.; Radix Ginseng is available from decoction pieces factory of German-Chinese medicine company of last Hisense; Rhizoma Zingiberis Recens is purchased in market; Acetaminophen, tamoxifen and tetramethyl azo azoles salt (MTT) are available from Sigma company; RPMI 1640 culture medium and calf serum are available from Gibco company; Penicillin and streptomycin are available from North China pharmaceutical Co. Ltd; Dimethyl sulfoxine (DMSO) is available from Chemical Reagent Co., Ltd., Sinopharm Group.

1.2 cell strain human hepatoma cell strain SMMC-7721 is available from Shanghai cell resource center of the Chinese Academy of Sciences, the normal liver cell is provided by auspicious moral hepatic disease institute (Shanghai).

1.3 instrument superclean bench (Su Jing group), CO2 gas incubator (Thermo Forma), inverted microscope (Chongqing Photoelectric Instruments Corp.), microplate reader (Shanghai Lei Bo Analytical Instrument Co., Ltd), ultrasonic washing instrument (Shanghai High Kudos Science Instrument Co., Ltd.).

1.4 the preparation of medical material decocting liquid and ultrasonic extraction liquid

1.4.1 the former side's decocting of Herba Sidae Rhombifoliae soup liquid (treatise on Febrile Diseases Zhang Zhongjing side) precision takes by weighing Radix Bupleuri 4.286g, Radix Scutellariae 3.214g, and Radix Ginseng 2.143g, Rhizoma Pinelliae Preparata 3.214g, Radix Glycyrrhizae Preparata 1.786g, Rhizoma Zingiberis Recens 3.214g, Fructus Jujubae 2.143g adds 10 times of water gaging 200ml, and infiltration is spent the night; Ebuillition of heated, little the boiling of little fire fried in shallow oil 1h, collects decocting liquid, and 4 layers of filtered through gauze get filtrate; The residue medicinal residues add 8 times of water gaging 160ml, get filtrate with aforementioned extracting method; Merging filtrate, the monolayer filter paper filtering, heating is concentrated into 20ml, and the microporous filter membrane through 0.45 μ m and 0.22 μ m filters respectively.

1.4.2 the Six-element side of subtracting decocting liquid Six-element (no Radix Bupleuri), Six-element (no Radix Scutellariae), Six-element (the no Rhizoma Pinelliae), Six-element (no Radix Ginseng), Six-element (no Radix Glycyrrhizae), Six-element (no Rhizoma Zingiberis Recens), 7 kinds of decocting liquid of Six-element (no Fructus Jujubae), the same 1.4.1 of medical material amount and method for making.

, fry in shallow oil the two flavor side of subtracting decocting liquid Radix Scutellariaes totally 6 kinds of decocting liquid, the same 1.4.1 of medical material amount and method for making 1.4.3 closing with Radix Bupleuri, the Rhizoma Pinelliae, Radix Ginseng, Radix Glycyrrhizae, Rhizoma Zingiberis Recens, Fructus Jujubae respectively.

1.4.4 single medical material decocting liquid Radix Bupleuri, Radix Scutellariae, the Rhizoma Pinelliae, Radix Ginseng, Radix Glycyrrhizae, Rhizoma Zingiberis Recens, 7 kinds of decocting liquid of Fructus Jujubae, the same 1.4.1 of medical material amount and method for making.

1.4.5 former side's ultrasonic extraction liquid precision takes by weighing 1.4.1 medical material amount, adds 5 times of water gaging 100ml, infiltration is spent the night; Ultrasonic 30min, 4 layers of filtered through gauze, the monolayer filter paper filtering, heating is concentrated into 20ml, and the microporous filter membrane through 0.45 μ m and 0.22 μ m filters respectively.

1.4.6 Radix Bupleuri ultrasonic extraction liquid medical material amount and the same 1.4.5 of method for making.

1.4.7 Radix Scutellariae ultrasonic extraction liquid medical material amount and the same 1.4.5 of method for making.

1.4.8 Radix Scutellariae+Radix Bupleuri ultrasonic extraction liquid medical material amount and the same 1.4.5 of method for making.

2. method and result

2.1 all experiment medicines final concentrations in 96 porocyte culture plates of drug solution are respectively 5,10,20,30,40mgml ^-1(being equivalent to former side's medical material concentration); Positive control is acetaminophen (AAP) (5mmolL ^-1); Negative control is PBS (phosphate buffer).

2.2 the cytoactive detection method adopts RPMI 1640 culture medium (to contain 10% calf serum, 100Uml ^-1Penicillin, 100 μ gml ^-1Streptomycin), with concentration 5 * 10 ⁴Ml ^-1Human liver cancer cell be inoculated in 96 orifice plates, every hole 200 μ l, in 37 ℃, 5%CO ₂Overnight incubation in the incubator.Discard on the plate original liquid in each hole, add the cell culture fluid that contains the respective concentration medicine respectively, 4 multiple holes of the every concentration of every medicine, in 37 ℃, 5%CO ₂Cultivate in the incubator.Behind the 24h, add MTT (5mgml ^-1) 20 μ l/ holes.37 ℃, 5%CO ₂After temperature is incubated 4h in the incubator, abandon on the plate original liquid in each hole, add DMSO 200 μ l/ holes and precipitate with Rong Xie Jia Za (formazan).After room temperature is put 30min, under the 570nm wavelength, measure each hole optical density value (OD) with microplate reader.Calculate cytoactive: cytoactive (cell viability, %)=experimental group optical density value/matched group optical density value * 100%.

2.3 homogeneity test of variance and t check are adopted in statistical disposition.Inspection level α=0.1 of homogeneity test of variance, inspection level α=0.05 of t check.

2.4 result

2.4.1 the former side's decocting of Herba Sidae Rhombifoliae soup liquid has remarkable inhibition to human hepatoma cell strain growth, the former side's decocting of Herba Sidae Rhombifoliae soup liquid 20,30,40mgml ^-1The cell relative activity of concentration group is respectively 66.3%, 50.1% and 35.1% (Fig. 1, table 1);

2.4.2 former side and Radix Bupleuri ultrasonic extraction liquid do not have remarkable inhibition, Radix Scutellariae, Radix Scutellariae+Radix Bupleuri ultrasonic extraction liquid have remarkable inhibition (Fig. 1, table 1);

2.4.3 in the Six-element side of the subtracting decocting liquid, the decocting liquid that does not contain Radix Scutellariae does not have remarkable inhibition, other the 6 kinds decocting liquid that contain Radix Scutellariae all have remarkable inhibition (Fig. 2, table 2);

2.4.4 in the single medicinal material decocting liquid, only Radix Scutellariae has remarkable inhibition, other 6 kinds of single medicinal materials all do not have remarkable inhibitory action (Fig. 2, table 2);

2.4.5 in the two flavor side of the subtracting decocting liquid, only Radix Scutellariae+Rhizoma Pinelliae closes decocting liquid does not have remarkable inhibition, other 5 kinds all have remarkable inhibitory action, wherein the inhibitory action of Radix Scutellariae+Radix Bupleuri is the strongest, the cell relative activity of its each concentration group all is lower than the cell relative activity that Radix Scutellariae, Radix Bupleuri are done the time spent respectively separately, prompting Radix Bupleuri may and strengthen the toxicity (Fig. 2, table 2) of Radix Scutellariae with the Radix Scutellariae synergism.

Table 1 mtt assay detect the 570nm optical density value (OD value) that the former side of Herba Sidae Rhombifoliae soup, Radix Bupleuri, Radix Scutellariae, Radix Scutellariae+Radix Bupleuri ultrasonic extraction liquid act on human hepatoma cell strain (x ± s, n=4)

Annotate: ^*P＜0.05, ^*P＜0.01

Table 2 mtt assay detect the 570nm optical density value (OD value) that the former side of Herba Sidae Rhombifoliae soup, the Six-element side of subtracting, the two flavor sides of subtracting and each single medical material decocting liquid acts on human hepatoma cell strain (x ± s, n=4)

Annotate: ^*P＜0.05, ^*P＜0.01

Embodiment 2. is the human liver cell toxicity vitro detection of the Herba Sidae Rhombifoliae soup of composition of prescription again

Again reduced the ratio of liver toxicity medical material Radix Scutellariae in the prescription of compatibility, increased the ratio of the Rhizoma Pinelliae, the proportion of composing of each medical material was as follows during new group was put:

2.143 parts of Radix Bupleuri, 1.107 parts of Radix Scutellariaes, 1.107 parts of Radix Ginsengs, 2.107 parts of the Rhizoma Pinelliaes, 1.107 parts in Rhizoma Zingiberis Recens, 0.893 part in Radix Glycyrrhizae, 1.536 parts in Fructus Jujubae.

Method among the employing embodiment 1 is carried out mtt assay and is detected again the effect of the Herba Sidae Rhombifoliae soup decocting liquid of compatibility to the human liver cell strain, the Herba Sidae Rhombifoliae soup that the result shows again compatibility weakens greatly than the toxicity of the former side's Herba Sidae Rhombifoliae soup of treatise on Febrile Diseases Zhang Zhongjing, has kept the original curative effect of Herba Sidae Rhombifoliae soup simultaneously.

Claims (2)

1. a Herba Sidae Rhombifoliae soup composition of prescription is characterized in that, the medical material and the part by weight thereof that comprise are as follows,

2.143 parts of Radix Bupleuri, 1.107 parts of Radix Scutellariaes, 1.107 parts of Radix Ginsengs, 2.107 parts of the Rhizoma Pinelliaes, 1.107 parts in Rhizoma Zingiberis Recens, 0.893 part in Radix Glycyrrhizae, 1.536 parts in Fructus Jujubae

2. Herba Sidae Rhombifoliae soup composition of prescription as claimed in claim 1 is characterized in that this prescription is to the perform toxic attenuation of liver.

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