法医学杂志

法医学杂志 ›› 2015, Vol. 31 ›› Issue (6): 417-421.DOI: 10.3969/j.issn.1004-5619.2015.06.001

• 论著 •    下一篇

HO-1对脂多糖诱导大鼠肝细胞内质网应激的作用

王艳莎，季英磊，王  涛，吴林琳，费成平，刘夷嫦，谷振勇   

  1. WANG Yan-sha, JI Ying-lei, WANG Tao, WU Lin-lin, FEI Cheng-ping, LIU Yi-chang, GU Zhen-yong
  • 发布日期:2015-12-25 出版日期:2015-12-28
  • 通讯作者: 谷振勇，男，博士，教授，主要从事创伤性MODS、血管生物学和呼吸肌疲劳信号转导机制研究；E-mail：zygusz@126.com
  • 作者简介:王艳莎（1987—），女，硕士，主要从事法医病理学研究
  • 基金资助:

    国家自然科学基金资助项目（81273341）；江苏省高校优势学科建设工程资助项目（PAPD）

Effects of HO-1 on Lipopolysaccharide-induced Endoplasmic Reticulum Stress of Rat Hepatocytes

南通大学医学院法医学系，江苏 南通 226001   

  1. Department of Forensic Medicine, Medical College of Nantong University, Nantong 226001, China
  • Online:2015-12-25 Published:2015-12-28

被引次数

1

摘要： 目的 探讨抗氧化蛋白血红素氧合酶-1（heme oxygenase-1，HO-1）对脂多糖（lipopolysaccharide，LPS）诱导大鼠肝细胞内质网应激的影响。 方法 大鼠正常肝细胞系BRL细胞培养，筛选有效HO-1 siRNA。用LPS、LPS+HO-1 siRNA、HO-1 siRNA和PBS溶剂分别处理大鼠肝细胞。用台盼蓝拒染实验检测细胞活力，Hoechst 33258荧光染色检测细胞凋亡，Western印迹法检测GRP78、CHOP、caspase-12以及HO-1蛋白表达。 结果 LPS能剂量依赖和时间依赖性诱导大鼠肝细胞HO-1蛋白表达上调，引起GRP78、CHOP和caspase-12蛋白表达增高，细胞活力降低和细胞凋亡率增高；HO-1 siRNA预处理明显抑制LPS对HO-1蛋白表达上调的诱导作用，并加剧LPS引起的内质网应激和细胞损伤。 结论 HO-1抑制LPS诱导大鼠肝细胞内质网应激介导的细胞损伤。

关键词: 法医病理学, RNA, 小分子干扰, 内质网, 应激, 脂多糖类, 血红素氧合酶-1, 肝细胞, 大鼠

Abstract:  Objective To investigate effects of antioxidant stress protein heme oxygenase-1 （HO-1） on lipopolysaccharide （LPS）-induced endoplasmic reticulum stress （ERS） of rat hepatocytes. Methods The BRL cells （rat hepatocyte cell line） were cultured. The hepatocytes were treated with LPS, LPS+HO-1 siRNA, HO-1 siRNA and PBS solution, respectively. The cell viability was measured by trypan blue exclusion test. The apoptosis cells were detected by the fluorescent dye Hoechst 33258. Expressions of GRP78, CHOP, caspase-12 and HO-1 were detected by Western blotting. Results LPS caused an increase of HO-1 protein expression of rat hepatocytes in a dose-dependent and time-dependent manner, a up-regulation of GRP78, CHOP and caspase-12, a decrease in cell viability, and an increase in apoptosis rate of hepatocytes. Pretreatment of HO-1 siRNA inhibited the up-regulation of LPS-induced HO-1, however, aggravated ERS and cellular injury. Conclusion HO-1 inhibites ERS-mediated cellular injury of rat hepatocytes induced by LPS.

Key words: forensic pathology, RNA, small interfering, endoplasmic reticulum, stress, lipopolysaccharides, heme oxygenase-1, hepatocytes, rats

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