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
陈丽 1谭程匀 1韦维1
作者信息
- 1. 广西中医药大学,广西南宁 530200
- 折叠
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.
Abstract
Objective To explore the effect of Anwei decoction containing serum on PD-1/PD-L1 signal pathway related fac-tors 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 con-taining 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 cho-sen 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Key words
Anwei decoction/MC cells/Immune escape/PD-1/PD-L1/Apoptosis引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据