PARP-1抑制剂5-AIQ对实验性结肠炎小鼠Treg/Th17的调节作用

Effect of PARP-1 inhibitor 5-AIQ regulating Treg/Th17 in experimental colitis mice

[目的]探讨聚腺苷二磷酸核糖聚合酶-1(PARP-1)抑制剂5-氨基异喹啉酮(5-AIQ)对葡聚糖硫酸钠(dextran sodium sulfate,DSS)诱导的实验性结肠炎小鼠Th17/Treg平衡的调节作用及其机制.[方法]将30只C57 BL/6 J小鼠随机分为3组:空白组、DSS组、5-AIQ组,每组10只;小鼠自由饮用质量浓度为30 g/L的DSS共7d,组建UC模型.空白组:正常饮水的同时给予0.9%氯化钠溶液,腹腔注射7 d;DSS组:造模同时给予0.9%氯化钠溶液,腹腔注射7 d;5-AIQ组:造模同时给予5-AIQ(1.5 mg/kg),腹腔注射7d.观察各组小鼠体质量、大便黏稠度及出血情况、计算结肠炎相关的疾病活动指数(disease activity index,DAI)和结肠组织病理评分,RT-PCR检测小鼠结肠组织中的炎症因子IL-10、IL-17、IL-21、IL-35,同时检测小鼠结肠组织中Th17和Tregs细胞分化相关转录因子RORγt、STAT3、Foxp3、Smad3表达.[结果]与DSS组相比,空白组和5-AIQ组中DAI降低(P<0.01);5-AIQ组结肠病理损伤明显改善;与空白组相比,DSS组IL-10、IL-35、Foxp3、Smad3蛋白下调,IL-17、IL-21、RORγt、STAT3蛋白上调;5-AIQ组与DSS组比较,IL-10、IL-35、Foxp3、Smad3蛋白上调,IL-17、IL-21、RORyt、STAT3蛋白下调,差异有统计学意义(P<0.01).[结论]PARP-1抑制剂5-AIQ可能通过调控实验性结肠炎小鼠Treg/Th17免疫平衡起到抑制肠道炎症,保护结肠黏膜的作用.

[Objective]To investigate the regulation of the poly(ADP-ribose)polymerase-1(PARP-1)inhibitor 5-aminoisoquinolinone(5-AIQ)on the balance of T helper cell 17(Th17)/regulatory T cells(Treg)in experimental colitis mice induced by dextran sodium sulfate(DSS)and its mechanism.[Methods]Thirty C57 BL/6J mice were randomly divided into 3 groups:control group(water+saline intraperitoneally),model group(DSS+saline intraperitoneally),5-AIQ group(5-AIQ 1.5 mg/kg,intraperitoneally),10 in every group.The mice were taken DSS(the mass concentration was 30 g/L)in drinking water for 7 days to es-tablish acute colitis model.The body mass,stool consistency and hemorrhage of every group were observed and the colitis related disease activity index(DAI)and colon histopathological score were caleulated.The inflammatory factors IL-10,IL-17,IL-21 and IL-35 in the colon were detected by RT-PCR.The expression of Th17 and Tregs differentiation related transcription factors RORyt,STAT3,Foxp3 and Smad3 were detected by Western blotting.[Results.]Compared with the model group,the DAIs of the control group and the 5-AIQ group were decreased(P<0.01).The pathological damage of colon was significantly improved after the intervention of 5-AIQ.Compared with the control group,the expressions of IL-10,IL 35,Foxp3,Smad3 were down-regulated;and the expressions of IL-17,IL-21,RORγt,STAT3 were up-regulated in the model group(P<0.01).After 5-AIQ treatment,compared with the model group,the expressions of IL:10,IL-35,Foxp3,Smad3 were up regulated,and the expressions of IL-17,IL-21,RORYt,STAT3 were significantly decreased(P<0.01).[Conclusion]The PARP-1 inhibitor 5-AIQ may inhibit intestinal inflammation and protect the colonic mucosa by regulating the Treg/Th17 immune balance in experimental colitis mice.