基础医学与临床 ›› 2014, Vol. 34 ›› Issue (7): 909-913.

• 研究论文 • 上一篇    下一篇

HMGB1-TLR4促进缺氧复氧介导的大鼠肾小管上皮细胞凋亡

尹金凤1,刘秀娟2,沈江山1,周阳1,尹秀英1   

  1. 1. 南昌大学第四附属医院
    2. 解放军第九四医院
  • 收稿日期:2013-10-21 修回日期:2014-05-19 出版日期:2014-07-05 发布日期:2014-06-24
  • 通讯作者: 刘秀娟 E-mail:xiujuan_liu796@163.com

HMGB1-TLR4 promote anoxia/reoxygenation injury-mediated apoptosis in rat proximal tubular epithelial cells

  • Received:2013-10-21 Revised:2014-05-19 Online:2014-07-05 Published:2014-06-24

摘要: 目的 研究缺氧复氧后大鼠肾小管上皮细胞中高迁移率族蛋白1(HMGB1) 和Toll样受体4(TLR4)的表达及其在肾小管上皮细胞凋亡中的调控机制 方法大鼠原代肾小管上皮细胞(PTECs) 置于6孔培养板培养,随机分为对照组(Control)、缺氧复氧组(A/R)、HMGB1抗体处理组(A/R+HMGB1)和TLR4抗体处理组(A/R+TLR4)(每组n = 3)。用RT-PCR检测HMGB1和TLR4 mRNA的表达;Western blot法检测HMGB1、TLR4 及凋亡相关分子Bcl-2、Bax、CHOP和caspase-8的蛋白表达;流式细胞术检测细胞凋亡。结果 A/R组HMGB1和TLR4的mRNA和蛋白的表达水平较对照组均显著上调 (P<0.01),细胞凋亡水平较对照组显著增加 (P<0.01),凋亡相关分子Bax、CHOP和caspase-8蛋白表达水平较对照组显著增加( P<0.01),Bcl-2蛋白表达较正常对照组下调(P<0.05)。 经抗体处理后,HMGB1抗体处理组和TLR4抗体处理组PTECs细胞凋亡水平较缺氧复氧组显著下降(P<0.05),凋亡相关分子Bax、CHOP和caspase-8蛋白表达水平较缺氧复氧组下降( P<0.05),Bcl-2蛋白表达变化不明显。结论 HMGB1-TLR4轴可促进缺氧复氧介导的大鼠肾小管上皮细胞凋亡。

关键词: 高迁移率族蛋白1, Toll样受体4, 细胞凋亡

Abstract: Objective To study the expression of high mobility group box-1 protein (HMGB1) and Toll-like receptor 4 (TLR4) in renal proximal epithelial cells following by anoxia/reoxygenation injury and their underlying roles in cell apoptosis. Methods Rat primary proximal tubule epithelial cells (PTECs) were cultured in six-well plates and randomly divided into Control, anoxia/reoxygenation group(A/R), HMGB1 antibody treatment group (A/R+HMGB1) and TLR4 antibody treatment group (A/R+TLR4). RT-PCR was used to detect the mRNA levels of HMGB1 and TLR4; Western blot was used to detect the protein expression of HMGB1, TLR4 and the apoptosis-related factors, such as Bcl-2, Bax,CHOP and caspase-8; Flow cytometry was used to detect apoptosis. Results The expression of mRNA and protein of TLR4 and HMGB1 increased significantly in A/R group compared with control (P<0.01). The apoptosis cells in A/R group increased significantly compared with control (P<0.01). The expression of Bax, CHOP and caspase-8 increased significantly compared with control (P<0.01). While the expression of Bcl-2 decreased compared with control (P<0.05). After HMGB1 and TLR4 antibodies treatment, the apoptosis cells in A/R+HMGB1 group and A/R+TLR4 group decreased significantly compared to A/R group (P<0.05). The expression of Bax, CHOP and caspase-8 also decreased significantly compared with A/R group (P<0.05). The change of Bcl-2 protein expression was not obviously. Conclusions HMGB1-TLR4 axis promote apoptosis of PTECs following A/R injury.

Key words: TLR4, HMGB1, Apoptosis