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