Effect Of Qinggan Huayu Granule On Anti - Tumor Effect And Immune Function, RAF / MEK / ERK Pathway In S180 And H22 Tumor Bearing Mice

Background:With the rapid development of social and economic life, the human disease spectrum changes.The cancer becomes one of the main reasons leading to human death, its morbidity and mortality are increasing year by year, and it seriously endangers human health.Hepatocellular carcinoma (HCC) is a common solid cancer and the third most frequent cause of cancer-related mortality worldwide. The 5-year relative survival rate for patients with HCC is only 7%, and very few patients with symptomatic disease survive for> 1 year. One of the primary reasons for the poor prognosis of patients with HCC is the lack of effective treatment options, especially for those with advanced disease. Although surgery and percutaneous ablation can achieve long-term control in some patients with early HCC, fewer than 40% of patients are diagnosed at early stages; hence, only a minority of HCC patients are eligible for potentially curative therapies,such as resection, transplantation, or percutaneous ablation. By the therapy of traditional Chinese medicine,it is not only can effective control progress,reduce the recurrence and metastasis, but also can improve the life quality of patients, prolong survival time, therefore the research of treatment primary liver cancer by traditional Chinese medicine has more and more attention.This study is supported by the national science and technology major projects, major new drug candidate drug research topic:QingGan HuaYu Particles in the treatment of primary liver cancer(2012ZX09103201-021). QingGan HuaYu Particles is an effective recipe in treatment of primary hepatic carcinoma,it is formed by more than years of clinical experience of Professor Yao Shukun,it is composed of Radix Scutellariae, flavescent sophora root, largehead atractylodes rhizome, zedoary, trigonous, Scutellaria barbata, Hedyotis diffusa, licorice and other drugs,it has the effect of heat-clearing and detoxifying, invigorating spleen for eliminating dampness,promoting blood circulation to remove blood stasis, soften hard.Preliminary study shows that Qinggan can be used in most cases of primary liver cancer patients, curative effect is exact, mechanism studies showed that it has action characteristics with multiple links, multiple target molecule network,it can improve immune function of patients, prolong survival time, improve the quality of life,and has the extremely value to further research.Objective:To study the anti-tumor effect and mechanism of QingGan HuaYu Particles on S180 and H22 hepatoma-bearing mice.Methods:The S180 and H22 hepatoma tumor-bearing model was established in mice,and mice were randomly divided into model group,fluorouracil group,PingXiao capsule, QingGan HuaYu Particles groups. Since modeling with appropriate drug intervention for 10 days. The proportion of CD3+、CD3+CD4+、CD3+CD8+、CD4+CD25+Tregs in serum were evaluated through Flow cytometric method on the 11 th day. The level of IL-10, TGF-β1 in serum were evaluated through ELASA.Then,the mice were killed to strip the tumors,liver and spleen,then calculate the inhibitory rates of tumor growth, the liver index and the liver index.The level of B-RAF、PTEN were evaluated with RT-PCT.The expression of VEGF. MEK、p-MEK protein were evaluated with Western blot.Results:Part Ⅰ:To study the anti-tumor effect and immune function of QingGan HuaYu Particles on S180 hepatoma-bearing mice.The inhibitory rate of Fluorouracil group,PingXiao capsule group, QingGan HuaYu Particles groups of dose 0.45g/kg、0.90g/kg、1.35g/kg、1.80g/kg、 2.25g/kg、2.70g/kg、3.6g/kg、4.5g/kgwere57%、29.0%、14.1%、17.3%、40.5%、36%、 17.2%、17.8%、12%、3% The body weight, the liver and spleen index of Fluorouracil group was significantly lower than the model group.The body weight, the liver and spleen index of PingXiao capsule group, QingGan HuaYu Particles groups were no significant compared with model group.Part Ⅱ:The inhibitory rate of Fluorouracil group,PingXiao capsule group, QingGan HuaYu Particles groups of dose 0.45 g/kg,0.90 g/kg,1.35 g/kg,1.80 g/kg,2.25 g/kg,2.70 g/kg were 57.1%、19.9%、11.5%、24.4%.29.9.6%、38.6%.28.3%、23.4%.The best does of QingGan HuaYu Particles were 1.80 g/kg.The body weight, the liver and spleen index of Fluorouracil group was significantly lower than the model group.The body weight, the liver and spleen index of PingXiao capsule group, QingGan HuaYu Particles groups were no significant compared with model group.Part Ⅲ:The proportion of CD3+CD3+CD4+and CD3+CD8+of QingGan HuaYu Particles groups were higher than the model group, but there was no statistically significant difference.The proportion of CD4+CD25+Treg of QingGan HuaYu Particles 1.80 g/kg, 2.70 g/kg groups were significantly lower than that of model group.There was no significant difference Between PingXiao capsule group,QingGan HuaYu Particles 0.90 g/kg dose group with model group.Our study also revealed a significant inverse correlation between the levels of Tregs and CD8+T cells in model group and all modeled groups.The level of IL-10 of QingGan HuaYu Particles groups were significantly lower than that of model group. QingGan HuaYu Particles can potently inhibit the tumor growth on H22 hepatoma-bearing mice,it probably play its role effectively through the inhibitory of the CD4+CD25+Tregs and IL-10.Part Ⅳ:QingGan HuaYu Particles can reduce the level of B-RAF m-RNA,lower the expression of VEGF, decrease the phosphorylation of MEK.Conclusion:QingGan HuaYu Particles can potently inhibit the tumor growth on S180 and H22 hepatoma-bearing mice,it probably play its role effectively through immune function and RAF/MEK/ERK signal transduction pathway.