P2RX7和组蛋白修饰变化在曲古抑菌素A缓解烟草烟雾暴露所致小鼠卵巢损伤中的作用

The role of P2RX7 and histone modification in trichostatin A alleviating mouse ovary injury induced by cigarette smoke exposure

目的探索P2RX7和组蛋白修饰变化在曲古抑菌素A(TSA)抑制NLRP3缓解烟草烟雾(CS)暴露所致的小鼠卵巢损伤过程中的作用。方法通过CS暴露30 d方法制备小鼠卵巢损伤模型,给予TSA进行治疗;HE染色观察各组小鼠卵巢组织形态;Western blot技术检测各组小鼠卵巢组织组蛋白去乙酰化酶(HDAC)1、HDAC2、NLR家族Pyrin域蛋白3(NLRP3)、P2RX7、Zeste同源物增强子2(EZH2)表达以及H3K4me1、H3K4me2、H3K27me3和H3K9ac总的修饰水平变化。结果与control组相比,CS组小鼠卵巢总体积减小,卵泡总数减少,卵巢皮质发生萎缩,TSA可有效抑制CS暴露引起的小鼠卵巢组织形态结构改变。CS组小鼠卵巢组织中HDAC1、HDAC2、NLRP3和P2RX7表达水平较control组明显升高,TSA有效抑制了CS暴露激活的小鼠卵巢组织HDAC1、HDAC2及焦亡相关蛋白NLRP3和P2RX7的表达(P<0.05)。CS暴露后小鼠卵巢组织总H3K4me1、H3K4me2和H3K9ac表达水平明显升高,TSA有效抑制了CS暴露诱导的小鼠卵巢组织总H3K4me1、H3K4me2和H3K9ac的表达上调(P<0.05)。结论P2RX7和组蛋白修饰变化参与CS暴露所致的卵巢NLRP3炎性小体的激活以及TSA缓解卵巢损伤的过程。

Objective To explore the role of P2RX7 and histone modification in trichostatin A(TSA)inhibiting NLRP3 and alleviating ovarian damage caused by cigarette smoke(CS)exposure in mice.Methods An mouse ovarian injury model was prepared by CS exposure twice daily for 30 days.TSA was injected intraperitoneally into CS-exposed mice on alternate days in the TSA-treated group.HE staining was used to observe the morphological changes of ovary after CS exposure.HDAC1,HDAC2,NLRP3,P2RX7 and EZH2 levels in ovary were detected by Western blot assay.Global modification levels of H3K4me1,H3K4me2,H3K9 ac and H3K27me3 were also assessed by Western blot assay.Results Compared with the control group,the total ovary volume and the total number of follicles were reduced in the CS group,and the ovarian cortex was atrophy.TSA can effectively inhibit the morphological and structural changes of the mouse ovarian tissue caused by CS exposure.The expression levels of HDAC1,HDAC2,NLRP3 and P2RX7 in ovarian were significantly higher in the CS group than those in the control group.TSA effectively restored the decreased ovarian volume,cortex atrophy and follicle reduction induced by CS exposure(P<0.05).Western blot results showed that TSA significantly inhibited the increased expression levels of NLRP3 and P2RX7 induced by CS exposure.Furthermore,TSA significantly inhibited expression levels of global H3K4me1,H3K4me2 and H3K9 ac modifications induced by CS exposure(P<0.05).Conclusion P2RX7 and histone modifications participate in the activation of NLRP3 inflammasome and the alleviation of TSA injury induced by CS exposure in model mice.