Mechanical Research On Jianpiliqi Herb In Interfering In Hepatoma Cells In Mouse Hca-F

Purpose:Primary hepatoma is one of the common malignant tumors in China. It influences human health seriously due to its rapid progress,high malignancy, bad prognosis and short life span.With the development of modern medical technology,the predominance of local therapy for tumor has already appeared. Up till now,the comprehensive evaluation of live Quality,live span and long-term effect is ambiguous yet.Hence the effect of whole mechanisms on the formation and development of tumor have been taken as the focus of researches. More and more studies have shown that Traditional Chinese Medicine could improve immune status and increase survival rate,and the method of Jianpiliqi has been widely used.Jianpiliqi herb is a kind of effective anti-tumor agents in our hospital,which consists of huangqi,renshen,baizhu,yunling,chenpi,shengma, chaihu,danggui and so on.Discussion of role of Jianpiliqi herb on hepatoma can provide experimental proof for treatment of hepatoma。Material and method:Pixuganyu animal model was successfully built,and ascites inducible hepatocarcinoma cell line Hca-F was chosen for tumor animal model.After solid tumor was built,animals were randomly divided into cancer bearing group,Jianpiliqi herb group,cyclophosphamide group and cyclophosphamide - Jianpiliqi herb combination group,and animals were intragastric administrated with normal sodium,Jianpiliqi herb, cyclophosphamide and Jianpiliqi herb plus cyclophosphamide for nine days.Then the effect of Jianpiliqi herb on morphology of hepatoma cells in mouse Hca-F was observed by light microscope(LM) and electron microscope(EM). Determination of lymphocyte subtypes CD4⁺,CD8⁺ and CD4⁺/CD8⁺was performed by FACS. Determination of activity of Mg²⁺-ATPase and G-6-Pase was performed by light microscope - enzyme cytochemistry staining.Determination of apoptotic cells was performed by Tunel;Determination of activity of telomerase was performed by in situ hybridization;and the expression of p53,BCL-2 and C-myc from tumor tissues was determined by immunohistochemistry。Results:1.Compared with cancer bearing group,experimental groups have following characteristics:the tumor ball and the cell volume were smaller,and the pathological karyokinesis were fewer.Chromatin condensed with high density, Lymphocytes infiltrated in interstitial and seldom tumor cells can be seen in capillaries.Surrounding necrosis area increased.2.The tumor inhibition ratio in Jianpiliqi herb group is 26.8%and 38.8%in cyclophosphamide - Jianpiliqi herb combination group.It shows that Jianpiliqi her can inhibit growth of tumor and the inhibitory effect appears obviously when Jianpiliqi herb combines with cyclophosphamide.3.Through FACS,CD4⁺ lymphocytes,CD4⁺/CD8⁺ raised and CD8⁺ lowered in treatment groups(P＜0.05),and there are no significant differences between Jianpiliqi herb group,cyclophosphamide - Jianpiliqi herb combination group and cyclophosphamide group.4.Through the observation of light microscope - enzyme cytochemistry staining, compared with cancer bearing group,activity of Mg²⁺-ATPase and G-6-Pase increased obviously in treatment groups,enzyme reaction particles enlarged and increased with higher density.In treatment groups,Mg²⁺-ATPase and G-6-Pase positive cells increased sharply compared with the cancer bearing group(P＜0.01) and there is no statistical difference among individual group.5.Through the observation of EM,we found the formation of apoptotic body.6.Through Tunel,increased apoptotic cells and high apoptotic index can be seen in treatment groups(P＜0.01) especially in cyclophosphamide - Jianpiliqi herb combination group,there is no significant difference on apoptotic index in Jianpiliqi herb group and cyclophosphamide group,and there is significant difference on apoptotic index in cyclophosphamide - Jianpiliqi herb combination group and Jianpiliqi herb group(P＜0.05).7.Through in situ hybridization,compared with the cancer bearing group, activity of telomerase reduced obviously in treatment groups(P＜0.05 or P＜0.01),and there is no statistical difference among individual group.8.Through immunohistochemistry,compared with the cancer bearing group,the expression of p53,BCL-2 and C-myc decreased obviously in treatment groups (P＜0.05 or P＜0.01),and there is no statistical difference among individual group.Conclusion:1.Jianpiliqi herb can inhibit the proliferation of hepatoma cells in mouse Hca-F。2.Jianpiliqi herb can decrease the activity of Mg²⁺-ATPase and G-6-Pase of hepatoma cells,and regulates lymphocyte subtypes CD4⁺,CD8⁺ and CD4⁺/CD8⁺in peripheral blood,hence improves immune status in mouse Hca-F。3.Jianpiliqi herb can reduce the activity of telomerase,and hence promotes apoptosis in mouse Hca-F.It appears synergistic effect when Jianpiliqi herb combines with cyclophosphamide.4.Jianpiliqi herb can degrade the expression of p53,BCL-2 and C-myc in mouse Hca-F。...