Xiao Chai Hu Tang And Its Anti-hepatoma Effect And Mechanism Of Drug Group Compatibility

It is very significant to illuminate theory of traditional Chinese medicine for studying classical formula of material fundament,compatibility rules and action mechanism by modern science and technology.The classical formula has some fine features,which its medication perfect,compatibility compact,effect outstanding,and is held in esteem by many doctors.So to select classical formula of fine compatibility and higher clinic value as key point is not only reasonable but maneuverability and demonstration as well.Xiaochaihu decoction(XCHD) being compatibility compact and combination ingenious possessed many kinds of effect of strengthening healthy qi to eliminate pathogens,relieving shaoyang disorder,disperse the depressed liver-energy and cholagogue,clear and logical interior and exterior yin-yang, concordance entrails.It was used to treating liver disease and liver cancer in clinic.According to modern pharmaco-study indicated:XCHD could markedly inhibit S180 tumor of mice.This topic planed to select XCHD inhibiting liver cancer as elementary point.Used H22 and S180 tumor model of mice to affirm antitumor action of XCHD.On that basis,according its formula compatibility reason to comparison observe antitumor action of XCHD and its different herbal formulation of component.Then we found that XCHD and active drug group (shen-zao-cao) could markedly inhibit tumor and have immunologic enhancement.Meanwhile, we investigated the different extraction method to gain part which its inhibiting tumor action.At last,we approached antihepatocarcinoma effect mechanism of XCHD by extraorgan blood-serum pharmacology,cytology and immunohistochemical method.This thesis was made of two parts.The first part mainly summarized XCHD study progress of experiment and liver disease clinic.As a whole,empirical study of XCHD refered to its chemistry,preparation,application and pharmaco-study:effect on liver and gall system, recirculating system,gastro intestinal system,genito-urinary system,immune system, neuro-endocrine system and antitumor,relieve fever,antiinflammatory,anti-infection action. Liver disease clinical research of XCHD refered to acu-chronic hepatitis,hepatic fibrosis and cirrhosis,liver cancer,adiposis hepatica,chole-stomach disease and depressive disorder etc.we found that XCHD adopted prescription,add-deduct and drug combination to promote effect and degrade toxic and side-effect.The second part was consisted of five researchs.Research 1:Pharmaceutical research of Xiaochaihu Decoction.Method:Used the HPLC to determine the index component of bupleurum chinense, Baical Skullcap Root,Radix Ginseng,Prepared Radix Glycyrrhizae.To determine saikoside-a, baicalin,panaxsaponin,glycyrrhizic acidcontent of XCHD by the HPLC which it can considerate preparation process of XCHD if it is reasonable.Result:Saikoside-a content of bupleurum,chinense is 3.3351mg/g,Saikoside-a content of XCHD is 0.7171mg/g.Baicalin content of Baical Skullcap Root is 10.867%,Baicalin content of XCHD is 33.717mg/g.; Panaxsaponin Rg₁ and panaxsaponin Re total contents of Radix Ginseng are 0.902%, Panaxsaponin Rb₁ content of Radix Ginseng is 0.592%,Panaxsaponin Rg₁ and panaxsaponin Re total contents of XCHD are 0.278mg/g,Panaxsaponin Rb₁ content of XCHD is 0.2069mg/g; Glycyrrhizin content of Prepared Radix Glycyrrhizae is 1.73%,Glycyrrhizin content of XCHD is 0.135 mg/g;Baicalin diversion of XCHD had exceeded 50%and arrived by 58.61%.Conclusion:Saikoside-a and Baicalin diversions of XCHD had exceeded 50%.Decoction preparation process of XCHD is rather reasonable.And these results suggest that pharmacologic action of XCHD have intimately relationship with these component.Research 2:Study on inhibiting liver cancer action to affirm of Xiaochaihu Decoction.Method:Transplanted tumor H22 liver cancer to mice for modelling of solid tumor and ascites tumor,then filling on high,moderate,low dose of Xiaochaihu decoction drag(27g,13.5g, 6.75 g·kg^-1),5-FU intraperitoneal injection(20 mg·kg^-1) as positive control.After being administered for 10 days,solid tumor model mice were observed cancer weight,body weight, spleen index and thymus gland index.Aseites tumor model mice were observed live span after model mice having died mice and stopped administer for studying dose-effect relationship.For furtthering to study on inhibiting liver cancer action to affirm of Xiaochaihu decoction. Transplanted tumor S180 tumor carneus to mice,then filling on moderatedose of Xiaochaihu decoction(13.5g·kg^-1) and detect cancer weight.Result:Growth of H22 mice liver cancer was markedly inhibited by high,moderate,low dose group,the inhibiting rates were by 49.66%, 48.52%,36.91%,optimal administration dose is moderate dose by analyzing.And three dose groups of XCHD could extend live time of mice which its were transplanted by ascites tumor, live span extended rates were by 23.59%,36.33%,26.96%.Moderate dose group of XCHD inhibiting rate was by 36.88%on S180 tumor carneus mice..To compare with normal group, Spleen index of H22 rumor-bearing mice was higher than that of blank control group(P＜0.01), thymus gland index and body weight had not obvious changed(P＞0.05),spleen index of three dose groups had evidently increased(P＜0.01),thymus gland index and body weight of 5-FU group had evidently decreased(P＜0.001).Conclusion:XCHD can markedly inhibit growth of H22 mice solid liver cancer and extend live span of H22 aseites tumor mice.The moderate dose of XCHD is the best dose by aggregate analysis.XCHD can also inhibit growth of S₁₈₀ solid tumor cameus mice,but its action is not better than on H22 liver cancer.These results cue us that XCHD probably have therapeutic action to aim directly at liver cancer.Research 3:Effect of Xiaochaihu Decoction and different herbal formulation of component on inhibiting H22 liver cancer in mice and enhancing the immune function.Method:Transplanted tumor H22 liver cancer to mice,filling on Xiaochaihu decoction the different herbal formulations of its components(Chinese Thorowax Root- Baical Skullcap Root(A),Fresh Ginger-Pinellia Tuber(B),Ginseng- Chinese Date - Licorice Root(C),A+B, A+C,B+C and A+B+C.),5-FU intraperitoneal injection(20 mg·kg^-1) as positive control.After being administered for 10 days,we determined cancer weight,body weight,spleen index and thymus gland index for studying dose-effect relationship.Then Xiaochaihu decoction group (13.5 g·kg^-1) and Ginseng-Chinese Date-Licorice Root apozem group(5 g·kg^-1) were given the medicines for 10 days.To observing the weight of tumor,body weight and the NK cell activity, proliferation of the T lymphocytes by MTT and IL-2 level by ELISA.Result:In the study of compatibility role,only inhibiting rates of Ginseng-Chinese Date-Licorice Root apozem group and Xiaochaihu decoction group exceed 30%,the inhibiting rates were by 41.55%and 43.65%. To compare with normal group,spleen index of the different herbal formulation groups had evidently increased(P＜0.01),thymus gland index of the different herbal formulation groups had significant decreased(P＜0.05),thymus gland index and body weight of 5-FU group had evidently decreased(P＜0.001).Comparing the model group,the NK cell activity,proliferation of the T lymphocytes and IL-2 level of Xiaochaihu decoction were remarkable increased (P＜0.001),the NK cell activity and IL-2 level of Ginseng-Chinese Date-Licorice Root group were remarkable increased(P＜0.05),the NK cell activity and proliferation of the T lymphocytes and IL-2 level of 5-FU group were remarkable decreased(P＜0.01);Comparing the normal control group,body weight of 5-FU group had evidently decreased(P＜0.05).Conclusion:Xiaochaihu decoction and Ginseng-Chinese Date-Licorice Root group can markedly inhibit growth of H22 mice solid liver cancer,and improve the immune function of tumor-bearing mice,its inhibiting effect probably closely related with improving rumor-bearing mice machine body immune function,there had a hint that compatibility Ginseng- Chinese Date -Licorice Root of strengthening body resistance was cores of inhibiting effect.Research 4:Contradistinction of different extraction process of Xiaoehaihu Decoction on inhibiting H22 liver cancer in mice.Method:Transplanted tumor H22 liver cancer to mice,filling on different extraction process of Xiaochaihu decoction drug(13.5 g·kg^-1),5-FU intraperitoneal injection(20 mg·kg^-1) as positive control.Atter being administered for 10 days,we determined cancer weight,body weight,spleen index and thymus gland index for studying best extraction process.Result: Growth of H22 mice liver cancer was markedly inhibited by decoction process,gradient alcohol process,water-alcohol process group,the inhibiting rates were by 32.35%,43.97%and 31.37% optimal process is gradient alcohol process by analyzing.To compare with normal group,spleen index of the different extraction process groups had evidently increased(P＜0.01),thymus gland index and body weight of 5-FU group had evidently decreased(P＜0.001).Comparing the normal control group,body weight of 5-FU group had evidently decreased(P＜0.001).Conclusion:Xiaochaihu decoction of decoction process,gradient alcohol process and water-alcohol process can markedly inhibit growth of H22 mice solid liver cancer.Gradient alcohol extraction process is the best process.But this process is too complex and its inhibiting rate is not far more than classical decoction process.All in all,classical decoction process has rather rationality. Research 5:Study of mechanism of action of Xiaochaihu Decoction on inhibiting liver cancerMethod:Transplanted tumor H22 liver cancer to mice,filling on Xiaochaihu decoction drug(13.5g·kg^-1),5-FU intraperitoneal injection(20 mg·kg^-1) as positive control.After being administered for 10 days,we determined cancer weight,body weight,detected TNF-αand VEGF of blood-serum by ELISA and detected Vascular Endothelial Growth Factor(MVD) and apoptosis rates of tumor tissue by immunohistochemical method.We prepared medicated blood-serum of XCHD which it was observed inhibitory action on H22 cells in culture fluid by MTT method.Result:Growth of H22 mice liver cancer was markedly inhibited by moderate dose group,the inhibiting rates were by 43.28%.To compare with analogue group,XCHD could increase markly TNF-αconcentration of mice blood-serum(P＜0.05) and degrade VEGF concentration of mice blood-serum(P＜0.05),but TNF-αconcentration of 5-FU group had evidently decreased(P＜0.05).XCHD could induced tumor apoptosis and markedly cut down MVD(P＜0.05).But medicated blood-serum of XCHD could not have any inhibitory action on H22 cells in culture fluid.Conclusion:XCHD can inhibit tumor growed of H22 mice and new vascularization which it concern of elevating immune function,increasing TNF-αconcentration,degrading VEGF concentration and inducing tumor apoptosis.This thesis had made system document research to find that XCHD had definite therapeutic on hepatopathy and liver cancer which is a key underlying pharmacologic action of XCHD.Our research results had also confirmed that XCHD had conclusive antitumor action.Compatibility study results indicated that XCHD inhibiting tumor action is the same as shen-zao-cao herb group.Mechanism of action research results hinted that XCHD inhibiting action were generated by immunological regulation,elevating activity of cell factor,inhibiting new tumor angiogenesis and inducing tumor cell necrosis.This research will provide,which modern study evidence for one formula more usage of traditional Chinese medicine,some study mode and experience for system studying of classical formula.Meanwhile,it will offer certain platform for developing and utilizating resources of classical formula.