养阴扶正方调控PD-1/PD-L1通路逆转T细胞耗竭的机制研究

Research on Mechanism of Regulating PD-1/PD-L1 Pathway by Yangyin Fuzheng Decoction(养阴扶正方)to Reverse T Cell Depletion

目的:探讨养阴扶正方调控PD-1/PD-L1通路逆转T细胞耗竭的机制。方法:免疫磁珠分选PD-L1^(+)Lewis肺癌细胞,建立PD-L1^(+)Lewis肺癌、Lewis肺癌C57BL/6鼠移植瘤模型,建模成功后,随机分为8组,分别予以养阴扶正方、阿特珠单抗、阿特珠单抗+养阴扶正方干预;绘制肿瘤生长曲线,流式细胞术检测荷瘤鼠外周血PD-L1^(+)CD_(8)^(+)T、PD-L1-CD_(8)^(+)T细胞以及瘤组织的CD_(4)^(+)CD_(25)^(+) Foxp^(+)T的表达,Western blot检测瘤组织PD-L1、PD-1、Tim3蛋白表达,RT-PCR检测瘤组织PD-L1、PD-1 mRNA表达。结果:PD-L1^(+)Lewis肺癌、Lewis肺癌移植瘤模型的各项指标无统计学差异;养阴扶正方可抑制PD-L1^(+)Lewis肺癌、Lewis肺癌生长,减少荷瘤鼠外周血PD-L1^(+)CD_(8)^(+)T数量,增加PD-L1-CD_(8)^(+)T细胞数量,减少瘤组织PD-L1、PD-1、Tim3蛋白表达以及瘤组织PD-L1、PD-1mRNA表达。结论:PD-L1^(+)Lewis肺癌、Lewis肺癌的生物学行为无差异,调控PD-1/PD-L1通路逆转T细胞耗竭是养阴扶正方抑制肺癌生长的机制之一。

Objective:To explore the mechanism of regulating the PD-1/PD-L1 Pathway by Yangyin Fuzheng Decoction to reverse T cell depletion.Metheds:PD-L1^(+) Lewis lung cancer cells were separated by munomagnetic Beads,lung carcinoma xenograft C57BL/6 mouse models of PD-l1^(+) Lewis lung cancer and Lewis lung were estab-lished.After successful modeling,random allocation,each group was respectively given Yangyin Fuzheng Decoction,Atezolizumab,Atezolizumab+Yangyin Fuzheng Decoction.Draw tumor growth curve,the expression of PD-1^(+) CD_(8)^(+)T,PD-1^(-)CD_(8)^(+)T cells in peripheral blood and CD_(4)^(+)CD_(25)^(+) Foxp^(+) T cell in tumor tissues was detected by Flow cytometry;the expressions of PD-1,PD-L1、Tim3 protein in tumor tissues were detected by Western Blot and the expressions of PD-1,PD-L1 mRNA in tumor tissues were deteced by RT-PCR.Results:There was no statistical difference in the indexes of PD-l1^(+) Lewis lung cancer and Lewis lung cancer xenograft mouse model.Yangyin Fuzheng Decoction could inhibit the growth of PD-1^(+) Lewis lung cancer and Lewis lung cancer,decrease the expression of PD-1^(+) CD_(8)^(+)T cells,increase PD-1^(-)CD_(8)^(+)T cells in peripheral blood and CD_(4)^(+)CD_(25)^(+) Foxp^(+)T cell in tumor tissues;reduce the expressiond of PD-1,PD-L1、Tim3 protein as well as PD-1,PD-L1 mRNA in tumor tissues.Conclusion:Regulating the PD-1/PD-L1 pathway to reverse T cell depletion is one of the mechanisms of Yangyin Fuzheng Decoction anti-lung cancer.