miR-150 Deficiency Protects against FAS-Induced Acute Liver Injury in Mice through Regulation of AKT.
miR-150 Deficiency Protects against FAS-Induced Acute Liver Injury in Mice through Regulation of AKT.
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DOI：
10.1371/journal.pone.0132734
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Wu T
中科院分区：
文献类型：
--
作者：
Chen W;Han C;Zhang J;Song K;Wang Y;Wu T
尽管 miR-150 参与免疫细胞分化和激活的调节，但 miR-150 是否参与肝脏生物学和疾病仍不清楚。本研究旨在通过使用野生型和 miR-150 敲除 (KO) 小鼠来探索 miR-150 在 LPS/D-GalN 和 Fas 诱导的肝损伤中的潜在作用。虽然敲除 miR-150 并没有显着改变 LPS/D-GalN 诱导的动物死亡和肝损伤，但它可以防止 Fas 诱导的肝损伤和死亡。在 miR-150 KO 小鼠中，Jo2 诱导的血清转氨酶、凋亡肝细胞、PARP 裂解以及 caspase-3/7、caspase-8 和 caspase-9 活性的增加均显着减弱。与野生型肝脏相比，Jo2 处理的 miR-150 KO 小鼠的肝组织表达更高水平的 Akt1、Akt2、总 Akt 以及 p-Akt(Ser473)。 Akt 抑制剂 V 预处理可逆转 Jo2 诱导的 miR-150 KO 小鼠肝损伤。与野生型小鼠的肝细胞相比，从 miR-150 KO 小鼠中分离的原代肝细胞在体外也显示出针对 Fas 诱导的细胞凋亡的保护作用（其特征是 PARP 裂解不太明显、核碎片较少和 caspase 激活较少）。在用 Akt1 或 Akt2 3'-UTR 报告构建体（有或没有 miR-150 结合位点突变）转染的肝细胞中进行荧光素酶报告测定，确定 Akt1 和 Akt2 作为 miR-150 的直接靶标。尾静脉注射含有 pre-miR-150 的慢病毒颗粒增强了 Jo2 诱导的 miR-150 KO 小鼠肝损伤。这些发现表明，miR-150 缺陷可通过调节 Akt 通路来防止 Fas 诱导的肝细胞凋亡和肝损伤。
Although miR-150 is implicated in the regulation of immune cell differentiation and activation, it remains unknown whether miR-150 is involved in liver biology and disease. This study was performed to explore the potential role of miR-150 in LPS/D-GalN and Fas-induced liver injuries by using wild type and miR-150 knockout (KO) mice. Whereas knockout of miR-150 did not significantly alter LPS/D-GalN-induced animal death and liver injury, it protected against Fas-induced liver injury and mortality. The Jo2-induced increase in serum transaminases, apoptotic hepatocytes, PARP cleavage, as well as caspase-3/7, caspase-8, and caspase-9 activities were significantly attenuated in miR-150 KO mice. The liver tissues from Jo2-treated miR-150 KO mice expressed higher levels of Akt1, Akt2, total Akt, as well as p-Akt(Ser473) compared to the wild type livers. Pretreatment with the Akt inhibitor V reversed Jo2-induced liver injury in miR-150 KO mice. The primary hepatocytes isolated from miR-150 KO mice also showed protection against Fas-induced apoptosis in vitro (characterized by less prominent PARP cleavage, less nuclear fragmentation and less caspase activation) in comparison to hepatocytes from wild type mice. Luciferase reporter assays in hepatocytes transfected with the Akt1 or Akt2 3’-UTR reporter constructs (with or without mutation of miR-150 binding site) established Akt1 and Akt2 as direct targets of miR-150. Tail vein injection of lentiviral particles containing pre-miR-150 enhanced Jo2-induced liver injury in miR-150 KO mice. These findings demonstrate that miR-150 deficiency prevents Fas-induced hepatocyte apoptosis and liver injury through regulation of the Akt pathway.