C1q和肿瘤坏死因子相关蛋白4(CTRP4)降低子痫前期大鼠滋养层细胞的caspase-1和IL-1β水平

C1q and tumor necrosis factor related protein 4(CTRP4) suppresses caspase-1/IL-1β inflammatory pathway in trophoblasts of rat models with preeclampsia

目的探讨C1q和肿瘤坏死因子相关蛋白4(CTRP4)对子痫前期大鼠胎盘滋养层细胞的影响。方法构建子痫前期大鼠模型,采集正常孕鼠和模型大鼠胎盘滋养层组织,运用实时定量PCR和Western blot法检测CTRP4、白细胞介素1β(IL-1β)和caspase-1 mRNA和蛋白表达水平;分离培养正常孕鼠和模型鼠滋养层细胞,在不同时间点,运用流式细胞术检测碘化丙啶和caspase-1双阳性(PI+caspase-1+)细胞(pyroptosis),运用实时定量PCR和Western blot法检测IL-1β和caspase-1表达水平;在模型大鼠滋养层细胞培养基中分别加入(0.5、5、15、25、50)ng/m L CTRP4重组蛋白或(10、20)ng/m L CTRP4蛋白中和抗体,处理72 h后检测pyroptosis细胞数目和caspase-1、IL-1β水平。结果子痫前期大鼠胎盘滋养层组织caspase-1、IL-1β表达增强,CTRP4表达水平下调;CTRP4重组蛋白处理体外培养的大鼠滋养层细胞可显著减少PI+caspase-1+细胞数量并降低caspase-1、IL-1β水平,而CTRP4蛋白中和抗体处理显著增加PI+caspase-1+细胞数量并增强炎症反应。结论 CTRP4可显著抑制caspase-1/IL-1β炎症调节通路的活性,并抑制子娴前期大鼠胎盘滋养层细胞的pyroptosis。

Objective To explore the effect of C1q/tumor necrosis factor related protein 4（CTRP4） on the placental trophoblasts of preeclampsia model rats.Methods Placental trophoblastic tissues were respectively collected from normal pregnant rats and model rats with preeclampsia,and then mRNA and protein expression levels of CTRP4,interleukin 1β（IL-1β）,and caspase-1 were detected with quantitative real-time PCR（qRT-PCR） and Western blotting.Primary placental trophoblasts were isolated from normal pregnant rats and model rats; at different time points,flow cytometry was used to detect the number of PI^＋caspase-1^＋pyroptotic cells; and qRT-PCR and Western blotting were used to detect expression levels of IL-1β and caspase-1.Final y,recombinant CTRP4 protein（at the doses of 0.5,5,15,25 or 50 ng/m L） or neutralizing CTRP4 antibody（at the doses of 10 or 20 ng/m L） were added into the medium of trophoblasts from model rats; after incubation for 72 h,the number of pyroptotic cells and the expression levels of IL-1β and caspase-1 were detected.Results Caspase-1/IL-1β inflammatory pathway was activated and CTRP4 expression was downregulated in placenta trophoblastic tissue from rats with preeclampsia.CTRP4 recombinant protein treatment significantly inhibited pyroptosis and the caspase-1 /IL-1β pathway in trophoblasts derived from rats with preeclampsia,while CTRP4 neutralizing antibody treatment had an opposite effect on pyroptosis and inflammation.Conclusion CTRP4 can significantly inhibit the activation of caspase-1/IL-1βinflammatory pathway,and suppress the pyroptosis of trophoblasts derived from rats with preeclampsia.