Hao Wei, Zhao Zhi-Hui, Meng Qing-Tao, Tie Mu-Er, Lei Shao-Qing, Xia Zhong-Yuan
Department of Anesthesiology, Renmin Hospital of Wuhan University, Wuhan, 430060, China.
Department of Anesthesiology, Inner Mongolia Autonomous Region People's Hospital, Huhhot, 010017, China.
Cell Biol Int. 2017 May;41(5):495-504. doi: 10.1002/cbin.10745. Epub 2017 Mar 7.
Propofol has been found to play an important role in hepatic ischemia/reperfusion (I/R) injury with the antioxidant effects. However, the molecular mechanism of propofol in hepatic I/R injury has not been fully understood. Male Sprague-Dawley rats were randomly assigned into Sham group, hepatic I/R group, and propofol treatment group. I/R injury was attained by ischemia for 1 h and reperfusion for 2 h. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity were detected. QSG-7701 cells were cultured in hypoxia condition for 15 h and then in reoxygenation condition for 6 h to imitate hypoxia/reoxygenation (H/R) injury in vitro. Real-time RT-PCR and Western blot were performed to determine the expression of miR-133a-5p and MAPK6. Luciferase reporter assay was used to determine the regulation of miR-133a-5p on MAPK6. Propofol significantly reduced the activities of serum AST and ALT induced by hepatic I/R injury in rats. Propofol increased the level of miR-133a-5p and decreased the expression of MAPK6 in vivo and in vitro. Luciferase reporter assay showed that MAPK6 was a target of miR-133a-5p. Knockdown of miR-133a-5p abrogated the effect of propofol on the upregulation of MAPK6 induced by H/R. MAPK6 overexpression promoted the cell apoptosis induced by H/R which could be attenuated by propofol. Finally, we found that miR-133a-5p reversed the protective effect of propofol in rats with hepatic I/R injury. Propofol showed protective roles for hepatic I/R injury in vivo and H/R injury in vitro, which involved with miR-133a-5p regulating the expression of MAPK6.
已发现丙泊酚通过抗氧化作用在肝缺血/再灌注（I/R）损伤中发挥重要作用。然而，丙泊酚在肝I/R损伤中的分子机制尚未完全明确。将雄性Sprague-Dawley大鼠随机分为假手术组、肝I/R组和丙泊酚治疗组。通过缺血1小时和再灌注2小时诱导I/R损伤。检测血清天冬氨酸转氨酶（AST）和丙氨酸转氨酶（ALT）活性。将QSG-7701细胞在缺氧条件下培养15小时，然后在复氧条件下培养6小时以在体外模拟缺氧/复氧（H/R）损伤。进行实时RT-PCR和蛋白质印迹法以测定miR-133a-5p和丝裂原活化蛋白激酶6（MAPK6）的表达。使用荧光素酶报告基因检测法测定miR-133a-5p对MAPK6的调控作用。丙泊酚显著降低大鼠肝I/R损伤诱导的血清AST和ALT活性。丙泊酚在体内和体外均增加miR-133a-5p水平并降低MAPK6的表达。荧光素酶报告基因检测显示MAPK6是miR-133a-5p的靶标。敲低miR-133a-5p消除了丙泊酚对H/R诱导的MAPK6上调的作用。MAPK6过表达促进H/R诱导的细胞凋亡，而丙泊酚可减弱这种作用。最后，我们发现miR-133a-5p逆转了丙泊酚对肝I/R损伤大鼠的保护作用。丙泊酚对体内肝I/R损伤和体外H/R损伤均具有保护作用，这与miR-133a-5p调节MAPK6的表达有关。
