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MiR-494通过激活PI3K/AKT通路减轻大鼠肝缺血—再灌注损伤

陈鑫培 苏松 周鹏程 罗德 刘向东 刘安定 李波

陈鑫培, 苏松, 周鹏程, 等. MiR-494通过激活PI3K/AKT通路减轻大鼠肝缺血—再灌注损伤[J]. 器官移植, 2019, 10(3): 295-301. doi: 10.3969/j.issn.1674-7445.2019.03.012
引用本文: 陈鑫培, 苏松, 周鹏程, 等. MiR-494通过激活PI3K/AKT通路减轻大鼠肝缺血—再灌注损伤[J]. 器官移植, 2019, 10(3): 295-301. doi: 10.3969/j.issn.1674-7445.2019.03.012
Chen Xinpei, Su Song, Zhou Pengcheng, et al. MiR-494 alleviates hepatic ischemia-reperfusion injury in rats by activating PI3K/AKT signaling pathway[J]. ORGAN TRANSPLANTATION, 2019, 10(3): 295-301. doi: 10.3969/j.issn.1674-7445.2019.03.012
Citation: Chen Xinpei, Su Song, Zhou Pengcheng, et al. MiR-494 alleviates hepatic ischemia-reperfusion injury in rats by activating PI3K/AKT signaling pathway[J]. ORGAN TRANSPLANTATION, 2019, 10(3): 295-301. doi: 10.3969/j.issn.1674-7445.2019.03.012

MiR-494通过激活PI3K/AKT通路减轻大鼠肝缺血—再灌注损伤

doi: 10.3969/j.issn.1674-7445.2019.03.012
基金项目: 

四川省科技厅应用基础项目 2014JY0068

详细信息
    作者简介:

    陈鑫培,男,1993年生,硕士研究生,住院医师,研究方向为肝移植,Email:807903650@qq.com

    通讯作者:

    苏松,男,1978年生,博士,副教授,硕士研究生导师,研究方向为肝移植,Email: 13882778554@163.com

  • 中图分类号: R617, R-332

MiR-494 alleviates hepatic ischemia-reperfusion injury in rats by activating PI3K/AKT signaling pathway

More Information
  • 摘要:   目的  探讨微小核糖核酸(miRNA)-494对肝缺血-再灌注损伤(HIRI)的影响及相关作用机制。  方法  24只雄性SD大鼠随机均分为4组(每组6只):假手术组行腹部手术而未行肝脏缺血-再灌注;HIRI组大鼠行部分肝缺血60 min后,再灌注6 h;HIRI+agomir-miR-494组在术前7 d内每日腹腔注射agomir-miR-494(20 μL);HIRI+agomir-NC组在术前7 d内每日腹腔注射等量的agomir-NC。采用逆转录聚合酶链反应(RTPCR)法检测各组肝组织中miR-494的信使核糖核酸(mRNA)表达水平。采用相关试剂盒检测各组的肝损伤指标和氧化应激指标表达水平。观察各组肝组织病理学改变。采用试剂盒检测各组大鼠肝组织中凋亡细胞数和胞质组蛋白相关DNA片段。采用免疫印迹法检测各组凋亡相关蛋白以及磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶(AKT)通路相关蛋白的表达量。  结果  HIRI+agomir-miR-494组大鼠肝组织miR-494的mRNA水平显著高于HIRI+agomir-NC组(P < 0.01)。HIRI+agomir-miR-494组的血清肝损伤指标以及血清氧化应激指标均显著低于HIRI+agomir-NC组(均为P < 0.01)。与HIRI+agomir-NC组相比,HIRI+agomir-miR-494组肝细胞坏死明显减少和细胞完整性提高(P < 0.05),TUNEL阳性细胞数量明显减少(P < 0.05)。HIRI+agomir-miR-494组的cleaved-多聚二磷腺苷核糖聚合酶(PARP)、cleaved-含半胱氨酸的天冬氨酸蛋白水解酶-3(Caspase-3)、Bax水平明显低于HIRI+agomir-NC组(均为P < 0.05)。HIRI+agomir-miR-494组大鼠DNA片段显著少于HIRI+agomir-NC组(P < 0.01)。HIRI+agomir-miR-494组大鼠肝脏p-AKT、p-哺乳动物雷帕霉素靶蛋白(mTOR)和p-p70S6K表达量明显高于HIRI+agomir-NC组(均为P < 0.05)。  结论  miR-494可以减轻大鼠HIRI,其作用机制与激活PI3K/ AKT信号通路有关。

     

  • 图  1  各组大鼠miR-494水平、氧化应激指标、凋亡相关指标和DNA片段化分析的比较

    A图为各组大鼠肝组织中的miR-494水平;B图为各组大鼠血清中氧化应激指标MDA、TOA和OSI的水平;C图为各组大鼠肝细胞凋亡相关蛋白cleaved-PARP、cleaved-Caspase-3、Bax的表达水平;D图为各组大鼠DNA片段化分析;与假手术组比较,aP < 0.05和bP < 0.01;与HIRI+agomir-NC组比较,cP < 0.01

    Figure  1.  Comparison of miR-494 levels, oxidative stress indicators, apoptosis related indicators and DNA fragmentation analysis of rats among each group

    图  2  各组大鼠肝组织病理学改变(HE,×200)

    A图为假手术组;B图为HIRI组;C图为HIRI+agomir-NC组;D图为HIRI+agomir-miR-494组

    Figure  2.  Comparison of liver histopathological changes of rats among each group

    图  3  各组大鼠肝组织凋亡改变(TUNEL,×100)

    A图为假手术组;B图为HIRI组;C图为HIRI+agomir-NC组;D图为HIRI+agomir-miR-494组

    Figure  3.  Liver tissue apoptosis changes of rats among each group

    图  4  各组大鼠肝组织PI3K/AKT信号通路蛋白表达水平的比较

    与假手术组比较,aP < 0.05和bP < 0.01;与HIRI+agomir-NC组比较,cP < 0.01

    Figure  4.  Comparison of the protein expression levels of PI3K/AKT signaling pathway indicators in the liver tissue of rats among each group

    表  1  各组大鼠肝损伤指标的比较

    Table  1.   Comparison of liver injury indicators of rats among each group(x±s)

    组别 n ALT(U/L) AST(U/L) LDH(IU/L) GDH(U/L)
    假手术组 6 49±12 61±11 113±15 661±76
    HIRI组 6 1 163±32a 1 325±25a 657±76a 1 985±89a
    HIRI+agomir-NC组 6 1 250±26 1 523±33 665±73 1 992±83
    HIRI+agomir-miR-494组 6 421±21b 532±23b 258±51b 1 013±82b
      与假手术组比较,aP < 0.01;与HIRI+agomir-NC组比较,bP < 0.01
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出版历程
  • 收稿日期:  2019-02-08
  • 网络出版日期:  2021-01-19
  • 刊出日期:  2019-05-15

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