Study On The Effect And Mechanism Of The Flavonoids From Tetrastigma Hemsleyanum Diels Et Gilg In Mice With Lewis Lung Cancer In Tumor Microenvironment

Objective Through the research of clover flavonoids on model mice bearing Lewis lung carcinoma tumor volume,tumor weight and MDSCs(myeloid derived suppressor cells),STAT3(signal transducer and activator of transcription 3,p-STAT3,VEGF,Bcl-2,Bax and effect on the expression of IL-6,to explore the relationship between the immune regulation of the tumor microenvironment and the IL-6/stat3 signaling pathway,and to provide a theoretical basis for the clinical application of the clover leaf.Methods ①Lewis Lung Mice models were established by inoculated subcutaneously way,and were divided into Tetrastigma hemsleyanum Diels et Gilg flavone high dose group,middle-dose group,low-dose group,NS group,normal group,each group 8 rats.24h after inoculation,the treatment groups was began to intragastric administration one time every day,the normal group was not treated,They were treated for 14 days.②Observe the change of survival condition and weight every day.Administration of tenth day of measuring tumor diameter,tumor volume calculation,observed the growth of tumors.The fourteenth day was executed,the tumor was weighed,and the tumor inhibition rate was calculated.③Eyeball blood taking,ELISA method was used to determine IL-6.④Tumor tissue was used to detect the expression of STAT3,p-STAT3,Bcl-2,Bax and VEGF by Immunohistochemical method.⑤The bone marrow was used to detect the expression of MDSCs Flow cytometry.⑥One-way ANOVA and Repeated Measure were used for statistical analysis of the corresponding sample.Results ① Weight:The body weight of the normal saline group was higher than that of the other mice,and the statistics were significantly different.There was no significant difference between the medicine groups.②Tumor volume:the volume size of the tumor:NS group>low-dose group>middle-dose group>high-dose group.③Tumor weight:The tumor weights were less than the normal saline group,but there was no significant difference between the low dose group and the normal saline group(P>0.05);Compared to the medicine group,low-dose group>middle-dose group>high-dose group,and two comparative differences(P<0.05).④ The tumor inhibition rate:the Flavonoids from Tetrastigma hemsleyanum Diels et Gilg in high dose group 58.64%,middle-dose group35.49%,low-dose group 13.26%.⑤Serum content of IL-6:From high to low is NS group、low-dose group、middle and high-dose group,Compared with the normal group,each model groups has a very significant difference(P<0.01),Compared with the NS group,the high and middle dose of the drug administration group had significant difference(P<0.01),Compared with middle and low dose groups,the high dose group was significantly different in the high dose group(P<0.01).⑥Immunohistochemical method was used to detect the expression of STAT3,p-STAT3,Bcl-2,Bax and VEGF:Compared with the NS group:Each group of STAT3,p-STAT3,Bcl-2,and VEGF levels were lower than the NS group,the high-dose group was the most obvious(P<0.01),Within the group,The inhibitory effect of high-dose group was the best,there was a significant difference(P<0.01);bax:Compared with the NS group,The content of Bax in each administration group was higher than that in the NS group,with significant difference(P<0.01),Within the group,The content of high dose group was the highest,and there was significant difference compared with middle and low dose group(P<0.01).⑦MDSCs in the bone marrow:Compared with the normal group,the MDSCs module in the bone marrow were markedly higher than those in the normal group,with significant difference(P<0.01).Compared with the NS group,The drug group compared with the NS group had significant difference(P<0.01).Within the group,high-dose group>middle-dose group>low-dose group.Conclusion ①the Flavonoids from Tetrastigma hemsleyanum Diels et Gilg have anti-tumor effect,With the increase of the dose,the inhibition effect is enhanced.②The content of IL-6 in serum of mice with Lewis lung cancer was decreased by the Flavonoids from Tetrastigma hemsleyanum Diels et Gilg,and it was positively correlated with the dosage.③the Flavonoids from Tetrastigma hemsleyanum Diels et Gilg can inhibit the expression of p-STAT3 and STAT3,down regulate Bcl-2 protein,up regulate Bax protein,reduce the ratio of bcl-2/bax and induce apoptosis of tumor cells,and reduce the expression of VEGF to inhibit angiogenesis.④The Flavonoids from Tetrastigma hemsleyanum Diels et Gilg in the leaves can decrease the proportion of MDSCs and reverse the immune escape of tumor.