安胃汤含药血清抑制MC细胞PD-1/PD-L1信号轴细胞免疫逃逸机制研究  

Mechanism of Anwei Decoction containing serum inhibiting cellular immune escape of PD-1/PD-L1 signal axis of MC cells

作　　者：陈丽[1] 谭程匀 韦维[1] CHEN Li;TAN Cheng-yun;WEI Wei(Guangxi University of Chinese Medicine,Nanning,Guangxi 530200,China)

机构地区：[1]广西中医药大学,广西南宁530200

出　　处：《时珍国医国药》2024年第4期827-831,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81960846)。

摘　　要：目的探讨安胃汤含药血清对MC细胞PD-1/PD-L1信号通路相关因子及免疫逃逸影响。方法以MC细胞为研究对象,设立空白组、安胃汤组、安胃汤^(+)阻断剂组、阻断剂组;CCK8检测12h、24h、48h细胞增殖情况,确定最佳检测时间点及含药血清比例;检测各组细胞上清液LDH活性;AO-EB染色法检测细胞膜的完整性;ELISA检测CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+)水平;Real-time PCR和Western-blot检测PD-1、PD-L1蛋白和基因的表达。结果选用48h时间点、10%含药血清比例加入后续实验;与其余三组比较,安胃汤组LDH活性增高(P<0.01);给药组AO-EB染色均可见细胞核形态改变;与其余各组比较,安胃汤组CD8^(+)表达下降(P<0.01)、CD4^(+)及CD4^(+)/CD8^(+)水平升高(P<0.01);安胃汤组与其余各组比较,PD-1、PD-L1蛋白和基因表达下降(P<0.01,P<0.05)。结论安胃汤能通过降低PD-1、PD-L1、CD8^(+)水平,升高CD4^(+)及CD4^(+)/CD8^(+)表达,促进细胞凋亡抑制细胞免疫逃逸,从而实现抗CAG作用。Objective To explore the effect of Anwei decoction containing serum on PD-1/PD-L1 signal pathway related factors and immune escape of MC cells[MNNG induced malignant transformation of human gastric mucosal epithelial cell line(GES-1)].Methods MC cells were divided into blank group,Anwei decoction group,Anwei decoction+blocker group and blocker group.CCK8 was used to detect cell proliferation at 12h,24h,and 48h,and determine the optimal detection time and drug containing serum ratio;to detect the LDH activity in cell supernatant of each group;AO-EB staining was used to detect the integrity of cell membrane;ELISA was used to detect the level of CD4~+,CD8~+,CD4~+/CD8~+;Real-time PCR and Western-blot was used to detect the expression of PD-1,PD-L1 protein and gene.Results 48 hours and 10%drug containing serum were chosen to join the follow-up experiment;compared with the other three groups,the activity of LDH in Anwei decoction group was higher(P<0.01);the morphological changes of nucleus were observed by AO-EB staining in the treatment group;compared with other groups,the expression of CD8~+in Anwei decoction group decreased and the levels of CD4~+and CD4~+/CD8~+increased(P<0.01);compared with other groups,the protein and gene expression of PD-1 and PD-L1 in Anwei decoction group decreased(P<0.01,P<0.05).Conclusion Anwei decoction can achieve anti-CAG effect by reducing the levels of PD-1,PD-L1 and CD8~+,increasing the expression of CD4~+and CD4~+/CD8~+,promoting apoptosis and inhibiting cellular immune escape.

关 键 词：安胃汤 MC细胞 免疫逃逸 PD-1/PD-L1 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]