The in vitro effects of phospholipase D1-mTOR axis in liver fibrogenesis
The in vitro effects of phospholipase D1-mTOR axis in liver fibrogenesis
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磷脂酶D1-mTOR轴在肝纤维化中的体外作用
DOI：
10.1016/j.lfs.2020.117595
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发表时间：
2020
期刊：
影响因子：
6.1
通讯作者：
Changqing Yang
中科院分区：
文献类型：
--
作者：
Yizhong Chang;Lu Xia;Meiyi Song;Min Tang;Bhuvanesh Kinish Patpur;Jing Li;Wenzhuo Yang;Changqing Yang
目的肝星状细胞(HSCs)的激活在肝纤维化进程中起核心作用。磷脂酶D(PLD)参与多种细胞活动。主要方法采用免疫印迹和免疫组织化学方法检测PLD诱导的小鼠肝纤维化模型肝内PLD1和PLD2的表达。用重组转化生长因子β-1(转化生长因子-β-1)诱导大鼠肝干细胞系(HSC-T6)建立肝纤维化细胞模型。从增殖、促纤维化标志物的表达和迁移等方面评价纤维化的发生。用小分子PLD1siRNA、雷帕霉素(mTOR抑制剂)和MHY1485(mTOR激活剂)处理HSC-T6细胞，观察PLD1mTOR轴在转化生长因子β1诱导的肝纤维化中的作用。用PLD抑制剂抑制PLD1活性或下调PLD1mRNA的表达可显著抑制转化生长因子β1诱导的肝纤维化形成，表现为细胞增殖减少和促纤维化标志物表达减少。此外，PLD 1抑制剂或PLD 1-siRNA均能显著抑制HSC-T6的mTOR活性。此外，在转化生长因子β-1诱导的肝纤维化中，PLD1mTOR抑制剂不仅具有与雷帕霉素相似的作用，而且还能钝化MHY1485促进HSC-T6细胞增殖的作用。
AimsThe activation of hepatic stellate cells (HSCs) plays a central role in liver fibrosis progression. Phospholipase D (PLD) enzymes participate in multiple cellular activities. However, whether and how PLD regulates HSCs activation remain elusive.Main methodsThe expression of intrahepatic PLD1 and PLD2 was determined in CCl4-induced mouse liver fibrosis models by western blot and immunohistochemistry. Cell model of liver fibrogenesis was constructed using rat HSCs line (HSC-T6) treated with recombinant transforming growth factor β1 (TGFβ1). Fibrogenesis was evaluated on the aspects of proliferation, expression of pro-fibrogenic markers and migration. The effects mediated by PLD1-mTOR axis on TGFβ1-induced fibrogenesis were evaluated using HSC-T6 treated with small-molecular PLD1 inhibitors, PLD1-SiRNA, rapamycin (mTOR inhibitor) and MHY1485 (mTOR activator).Key findingsSignificant increase of PLD1, not PLD2 was documented in CCl4-induced cirrhotic compared to normal liver tissues. Suppression of PLD1 activities by PLD inhibitors or down-regulation of PLD1 expression in HSC-T6 could significantly restrain TGFβ1-induced fibrogenesis, as reflected by decreased cell proliferation and reduced expression of pro-fibrogenic markers. Besides, either PLD1 inhibitor or PLD1-SiRNA significantly inhibited mTOR activity of HSC-T6. Moreover, PLD1 inhibitors not only exhibited similar effects with rapamycin in TGFβ1-induced fibrogenesis, but also blunted MHY1485 enhanced cell proliferation of HSC-T6.SignificanceThe PLD1-mTOR axis of HSCs could be therapeutically targeted in advanced liver fibrosis.