Study On The Mechanism Of Yiqi Jianpi Anti-cancer Prescription By Regulating Treg And Th17 Cells Enhancing Chemotherapy Effect Of Spleen-deficiency Colorectal Cancer Mice

Objective:To study the effect and mechanism of yiqi Jianpi Anti-cancer prescription on enhancing the chemotherapy effect of spleen-deficiency colon cancer mice。Methods:40 SPF male BALB/c mice were selected,randomly selected 6as healthy groups(natural growth,no treatment was done),the rest of the mice were established by overwork+hunger+bitter cold diarrhea method to establish a temper deficiency model。Then subcutaneously inoculated with CT-26 mice colon cancer cells for molding after the tumor body grew,the tumor-bearing mice were randomly divided into control group(this was tumor-bearing control group,which used normal saline gassing and intraperitoneal injection every day),the Chinese herbology group(only use Chinese herbology intragastriced administration),chemotherapy group(intraperitoneal injection using FOLFOX regimen),combined medication group(Yiqi Jian Spleen Anti-Cancer prescription+FOLFOX).After 14 days of drug intervention in each group,the general situation of each group of mice was observed,the tumor body mass and tumor volume of tumor-bearing mice were detected,and the tumor suppression rate was calculated;the apoptosis in each group of tumors was detected by Tunel staining method;the organ index of the main organs of each group of mice was calculated,and the histopathological changes of the main organs were observed by HE staining;the number of blood cells and the level of hemoglobin(Hb)in the blood of mice were detected by the automatic blood analyzer;The CD4~+T cells and Th17 cells in tumors and spleens were detected by flow cytometry.The expression of Treg cells was measured by ELISA to detect the levels of IL-17,IL-10,IFN-γ,TGF-βin tumors,and the expression of Ki-67,PCNA,RORγt and FOXP3 proteins in tumors were detected by immunohistochemistry.Results:Spleen deficiency mice was successfully established.Compared with the control group,Chinese herbology group,chemotherapy group and combination drug group had different degrees of inhibition effect on the tumor quality and tumor volume of tumor-bearing mice(P<0.05),and the tumor suppression rates of the three groups were:23.56%,48.98%,and61.91%,respectively,in the three groups,the tumor suppression effect of combination drug group is the best.Immunohistochemistry showed that compared with the control group,the relative expression of Ki-67 and PCNA proteins in the tumor tissues of the Chinese herbology Group,Chemotherapy Group and Combination Group reduced,statistically significant(P<0.01)compared with the chemotherapy group,the relative expression of Ki-67 and PCNA proteins in the combination group lower,and the difference between the two groups was statistically significant(P<0.05).TUNEL apoptosis staining showed that compared with the control group,tumor cell apoptosis significantly increased in the chemotherapy group and combination therapy group(P<0.01).In terms of organ index,compared with the healthy group,the spleen index of mice in each treatment group was increased to varying degrees(P<0.01),and the spleen index of the combination group was reduced(P<0.05).The blood routine showed that compared with the chemotherapy group,the red blood cells in the combination group increased and the difference was statistically significant(P<0.05);hemoglobin and white blood cells were elevated,but the difference between the two groups not obvious and there no statistical significance(P>0.05).HE staining showed that the chemotherapy group can be seen cardiological damage,and no obvious pathological damage was seen in the heart,liver,kidney and lung tissues of the remaining groups.Flow cytometry showed that the proportion of CD4~+T cells in the spleen of the Chinese medicine group and the combination group increased significantly compared with the control group(P<0.01),while the difference between Treg and Th17 cells not statistically significant(P>0.05).In the tumor tissues,compared with the chemotherapy group,the proportion of CD4~+T cells and Th17 cells increased,and Treg/Th17 decreased in the combination drug group,and the difference was statistically significant(P<0.05),and the proportion of Treg cells were decreased.ELISA showed that compared with the chemotherapy group,the content of IL-17 and IFN-γin the tumor tissues of the Chinese herbology group and the combination group was significantly increased,and the difference statistically significant(P<0.01).Compared with the chemotherapy group,the contents of IL-10 and TGF-βin the combination group were significantly reduced,and the difference was statistically significant(P<0.05).Immunohistochemistry showed that compared with the control group and the chemotherapy group,the relative content of RORγt protein in the tumor tissues of the Chinese herbology group and the combination group increased significantly,and the relative content of FOXP3protein decreased significantly,and the difference significantly significant(P<0.01).Conclusion:Yiqi Jianpi Anti-cancer Prescription can enhance the chemotherapy efficacy of spleen-deficiency colon cancer mice,inhibit the proliferation of tumor cells,promote the apoptosis of tumor cells,and its mechanism may be to improve the tumor microenvironment by increasing the proportion of Th17 cells in tumor tissue and reducing the proportion of Treg cells.