内异停对子宫内膜异位症大鼠PD-1/PD-L1途径及Th17/Treg表达的影响

Effects of Neiyiting on regulating Th17/Treg expression imbalance in endometriosis through the PD-1/PD-L1 pathway

目的:探讨内异停通过PD-1/PD-L1途径干预子宫内膜异位症大鼠Th17/Treg表达失衡的免疫机制。方法:45只雌性SD大鼠随机分为空白对照组、模型组、PD-1抗体组、内异停组、内异停低剂量组,每组9只。空白对照组不给予任何处理,其余各组大鼠均进行子宫内膜自体移植,术后28d,每组随机取1只大鼠开腹观察异位内膜生长情况,其余大鼠给药2周。ELISA检测外周血细胞因子IL-17A、IL-22、IL-10、TGF-β的表达。Western blot检测异位内膜PD-1、PD-L1、Foxp3、RoR-γt的表达。结果:术后28d可见移植部位形成典型的半球形囊状小泡。与空白对照组比较,模型组大鼠外周血中IL-17A、IL-22和异位内膜中PD-1、PD-L1、Foxp3表达显著升高(P<0.01),外周血中IL-10、TGF-β和异位内膜中ROR-γt表达显著降低(P<0.01);与模型组比较,内异停组、内异停低剂量组及PD-1抗体组外周血中IL-17A、IL-22表达显著降低(P<0.01),IL-10、TGF-β表达显著升高(P<0.01),内异停组及PD-1抗体组异位内膜中PD-1、PD-L1、Foxp3表达显著降低(P<0.05,P<0.01)。结论:内异停治疗子宫内膜异位症的作用机制可能与其阻断PD-1/PD-L1途径从而调节Th17/Treg失衡有关。

Objective:To explore the immune mechanism of the intervention effect of Neiyiting on the imbalance of Th17/Treg expression in endometriosis rats via PD-1/PD-L1 pathway.Methods:A total of 45 female SD rats were randomized to control group,endometriosis model group,PD-1 antibody group,Neiyiting group,and low-dose Neiyiting group,9 rats each group.No treatment was given to the control group,and all the other groups were performed autologous transplantation of endometrium.For each group,one rat was randomly selected and performed laparotomy 28 days after the auto transplantation for observing the growth of ectopic endometrium.Other rats were administrated with drugs for 2 weeks.The expressions of cytokines IL-17 A,IL-22,IL-10 and TGF-βin peripheral blood were detected by ELISA.The expressions of PD-1,PD-L1,Foxp3 and ROR-γt in ectopic endometrium were detected by Western blot.Results:Typical hemispherical cystic vesicles were observed at the graft site upon laparotomy 28 days after autologous transplantation.Compared with the control group,the expressions of IL-17 A,IL-22 in peripheral blood and PD-1,PD-L1 and Foxp3 in ectopic endometrium in model group increased significantly(P<0.01),while the expressions of IL-10,TGF-βin peripheral blood and ROR-γt in ectopic endometrium significantly decreased(P<0.01).Compared with the model group,the expressions of IL-17 A and IL-22 significantly decreased in Neiyiting group,lowdose Neiyiting group and PD-1 antibody group(P<0.01),while the expressions of IL-10 and TGF-βincreased significantly(P<0.01).The expressions PD-1,PD-L1 and Foxp3 significantly decreased in Neiyiting group and PD-1 antibody group(P<0.05,P<0.01).Conclusion:The mechanism of Neiyiting for treating endometriosis might be related to the regulation of Th17/Treg imbalance through blocking the PD-1/PD-L1 signal pathway.