Abstract：
Clonorchis sinensis is an important food-borne zoonotic parasite that is highly associated with liver fibrosis and cholangiocarcinoma. Further understanding of the pathogenesis of C. sinensis, especially liver fibrosis, could help us develop novel strategies for controlling clonorchiasis. Poly (ADP-ribose) polymerase-1 (PARP-1) can induce cellular parthanatos which is reported to be involved in liver fibrosis. Currently, whether C. sinensis could activate PARP-1 signaling to induce parthanatos or whether parthanatos play a role in C. sinensis-induced liver fibrosis is not clear. In the present study, the expression of PARP-1 and parthanatos indicators were detected in C. sinensis-infected mouse liver and in human intrahepatic biliary epithelial cells (HiBEpiCs) incubated with excretory/secretory products (ESPs) of C. sinensis. To explore the role of PARP-1 in C. sinensis infection, PARP-1 inhibitor NMS-P118 was used to block PARP-1 expression in vivo and vitro. The mortality rate, body weight, worm load, liver and bile duct lesions as well as PARP-1 and parthanatos indicators in C57BL/6 mice infected with C. sinensis, or in HiBEpiCs incubated with C. sinensis ESPs and NMS-P118 were analyzed and compared to the group without NMS-P118. The results showed that C. sinensis infection induced the activation of PARP-1 signaling as well as the translocation of AIF and MIF into the nucleus in mouse liver. ESPs of C. sinensis could induce PARP-1 up-regulation, ATP depletion and DNA damage in HiBEpiCs, indicating that C. sinensis could induce parthanatos. Inhibiting PARP-1 with NMS-P118 significantly reduced liver fibrosis and the number of larvae, increased the survival rate and body weight gain of the mice infected with C. sinensis. In addition, NMS-P118 decreased the expression of PARP-1 and alleviated ATP depletion as well as DNA damage in HiBEpiCs incubated with ESPs of C. sinensis. Our data indicated that C. sinensis and its ESPs could activate PARP-1 signaling to induce cellular parthanatos. NMS-P118 treatment alleviated liver fibrosis and promoted survival of the mice by inhibiting PARP-1, which suggested that PARP-1 could be used as a potential therapeutic target against clonorchiasis.
摘要：
华支睾吸虫是一种重要的食源性人畜共患寄生虫，与肝纤维化和胆管癌密切相关。进一步认识中华毛虫的发病机理,尤其是肝纤维化,可以帮助我们开发控制华支睾吸虫病的新策略。聚（ADP-核糖）聚合酶-1（PARP-1）可以诱导细胞parthanatos，据报道与肝纤维化有关。目前,尚不清楚中华绒螯蟹是否可以激活PARP-1信号以诱导parthanatos或parthanatos是否在中华绒螯蟹诱导的肝纤维化中起作用。在本研究中,PARP-1和parthanatos指标的表达在C.sinensis感染的小鼠肝脏和与C.sinensis的排泄/分泌产物（ESPs）孵育的人肝内胆管上皮细胞（HibepiCs）中检测。探讨PARP-1在中华绒螯蟹感染中的作用,PARP-1抑制剂NMS-P118用于体内和体外阻断PARP-1表达。死亡率,体重,蜗杆载荷,C57BL/6小鼠感染中华绒螯蟹的肝脏和胆管病变以及PARP-1和parthanatos指标，或在与中华梭菌ESPs和NMS-P118孵育的HibepiCs中进行分析并与没有NMS-P118的组进行比较。结果表明，中华绒螯蟹感染可诱导PARP-1信号的激活以及AIF和MIF在小鼠肝脏中的转位。华夏草的ESPs可以诱导PARP-1上调，HibepiCs中的ATP消耗和DNA损伤，表明C.sinensis可以诱导parthanatos。用NMS-P118抑制PARP-1显著降低肝纤维化和幼虫数量,提高了感染C.sinensis的小鼠的存活率和体重增加。此外,NMS-P118降低了PARP-1的表达，减轻了与C.sinensis的ESPs孵育的HibepiCs中的ATP消耗以及DNA损伤。我们的数据表明，C.sinensis及其ESP可以激活PARP-1信号传导以诱导细胞parthanatos。NMS-P118治疗通过抑制PARP-1减轻肝纤维化并促进小鼠的存活，这表明PARP-1可以用作对抗华支睾吸虫病的潜在治疗靶标。
