摘要
目的 探究雷公藤甲素(TPL)对MGC-803胃癌荷瘤小鼠肿瘤生长、免疫微环境及TLR4/NF-κB信号通路的影响.方法 将MGC-803细胞皮下注射至BALB/c雄性小鼠中,并随机分为模型组、阳性药物组、TPL-L组、TPL-H组、TPL-H+LPS组.检测小鼠肿瘤生长情况及脾脏和胸腺指数;流式细胞术检测小鼠外周血中CD4+T、CD8+T的水平;ELISA检测小鼠外周血中肿瘤坏死因子-α(TNF-α)、白介素-2(IL-2)、干扰素-Y(INF-γ)、白介素-4(IL-4)的水平;Western Blot检测小鼠肿瘤组织中TLR4/NF-κB通路相关蛋白的表达.结果 与模型组比较,TPL组胃癌小鼠肿瘤重量和体积减小,抑瘤率、脾脏和胸腺指数、外周血中CD4+T、CD4+/CD8+比例、TNF-α、IL-2、INF-γ水平升高(P＜0.05),CD8+T、IL-4水平及Toll样受体4(TLR4)、髓样分化因子(MyD88)、肿瘤坏死因子受体相关蛋白6(TRAF6)、核因子-KB(NF-κB)p65、细胞程序性死亡-配体1(PD-L1)蛋白表达降低(P＜0.05);TLR4的激活剂LPS进行回补实验显示,LPS逆转了TPL对胃癌小鼠肿瘤生长及免疫逃逸的抑制作用(P＜0.05).结论 TPL能够抑制MGC-803胃癌荷瘤小鼠的肿瘤生长,增强细胞免疫功能,抑制TLR4/NF-κB信号通路的激活.
Abstract
Objective To investigate the impacts of triptolide(TPL)on tumor growth,immune microenvironment and TLR4/NF-κB signaling pathway in MGC-803 gastric cancer bearing mice.Methods MGC-803 cells were injected subcutaneously into BALB/c male mice,and the mice were randomly divided into model group,positive drug group,TPL-L group,TPL-H group,and TPL-H+LPS group.The tumor growth and spleen and thymus indices of mice were detected;flow cytometry was applied to detect the levels of CD4+T and CD8+T in peripheral blood of mice;ELISA was applied to detect the levels of tumor necrosis factor-α(TNF-α),interleukin-2(IL-2),interferon-γ(INF-γ),and IL-4 in the peripheral blood of mice;Western Blot was applied to detect the expression of TLR4/NF-κB pathway related proteins in tumor tissues of mice.Results Compared with model group,the tumor weight and volume of gastric cancer mice in TPL group were lower,the tumor inhibition rate,spleen and thymus indexes,the levels of CD4+T,the proportion of CD4+/CD8+,the levels of TNF-α,IL-2,and INF-γ in peripheral blood were higher(P＜0.05),the levels of CD8 T and IL-4,the expression of TLR4,MyD88,TRAF6,NF-κB p65,and PD-L1 proteins in tumors was lower(P＜0.05);TLR4 activator LPS for compensatory experiments showed that LPS reversed the inhibitory effects of TPL on tumor growth and immune escape in gastric cancer mice(P＜0.05).Conclusion TPL can inhibit tumor growth,enhance cellular immune function,and inhibit the activation of TLR4/NF-κB signaling pathway in MGC-803 gastric cancer bearing mice.
基金项目
河北省医学科学研究课题计划项目(20221814)
NETL
NSTL
万方数据