法医学杂志

法医学杂志 ›› 2014, Vol. 30 ›› Issue (1): 13-18.DOI: 10.3969/j.issn.1004-5619.2014.01.003

• 论著 • 上一篇    下一篇

内质网应激参与介导脂多糖诱导的大鼠肝细胞凋亡

季英磊，闫  骏，王艳莎，刘夷嫦，谷振勇   

  1. （南通大学医学院法医学系，江苏 南通 226001）
  • 发布日期:2014-02-25 出版日期:2014-02-28
  • 通讯作者: 谷振勇，男，教授，博士研究生导师，主要从事创伤性MODS、血管生物学和呼吸肌疲劳的信号转导机制等研究；E-mail：zygusz@126.com
  • 作者简介:季英磊（1988—），女，山东临沂人，硕士研究生，主要从事创伤性MODS研究；E-mail：jiyinglei2006@126.com
  • 基金资助:

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

Endoplasmic Reticulum Stress Mediates Lipopolysaccharide-induced Apoptosis in Rat Hepatocyte

JI YING-LEI, YAN JUN, WANG YAN-SHA, LIU YI-CHANG, GU ZHEN-YONG   

  1. （Department of Forensic Medicine, Medical College of Nantong University, Nantong 226001, China）
  • Online:2014-02-25 Published:2014-02-28

被引次数

10

摘要： 目的 探讨内质网应激在脂多糖（lipopolysaccharide，LPS）诱导的肝细胞凋亡中的作用。 方法 肝细胞系BRL细胞培养，用LPS、内质网应激诱导剂毒胡萝卜素（thapsigargin，TG）、内质网应激抑制剂4-苯基丁酸（4-phenylbutyric acid，4-PBA）分别或组合处理肝细胞。用MTT比色法检测细胞活力的变化，Heochst 33258荧光染色检测细胞凋亡形态的变化，Annexin V-FITC/PI双染流式细胞术检测细胞凋亡率，Western印迹法检测内质网应激标志蛋白GRP78和内质网应激相关的凋亡蛋白CHOP、caspase-12和激活型caspase-3的表达。 结果 LPS引起肝细胞活力降低、细胞凋亡率明显增加，并具有量效和时效关系，LPS诱导GRP78、CHOP、caspase-12和激活型caspase-3的表达上调；TG可引起肝细胞活力降低和细胞凋亡增加，并能加重LPS引起的肝细胞损伤；4-PBA明显抑制LPS引起的肝细胞凋亡增加。 结论 内质网应激参与介导了LPS引起的肝细胞凋亡，提示内质网应激是LPS诱导肝细胞损伤的重要发病途径。

关键词: 法医病理学, 内质网, 脂多糖类, 肝细胞, 细胞凋亡, 大鼠

Abstract: Objective To investigate the role of endoplasmic reticulum stress （ERS） in lipopolysaccharide （LPS）-induced hepatocyte apoptosis. Methods Cells of the rat hepatocyte line BRL were cultured. The hepatocytes were treated with LPS, ERS inducer thapsigargin （TG）, and ERS inhibitor 4-phenylbutyric acid （4-PBA）, respectively or in their different combination. The cell viability was measured by MTT assay. The cyto-nuclear morphological changes of apoptosis cells were detected by the fluorescent dye Hoechst 33258. The apoptosis rate was assessed by flow cytometry with Annexin V-FITC/PI double-staining. Expressions of GRP78 as ERS marker protein, CHOP, caspase-12 and cleaved-caspase-3 as ERS related protein were detected by Western blotting. Results LPS could cause a decrease in cell viability and an increase in apoptosis rate in a dose- and time-dependent manner. The expression of GRP78, CHOP, caspase-12 and cleaved-caspase-3 proteins were significantly increased with LPS treatment. TG led to a marked decrease in cell viability and an increase in apoptosis rate, which aggravated the hepatocyte injury induced by LPS; whereas 4-PBA alleviated LPS-induced apoptosis. Conclusion ERS mediates LPS-induced hepatocyte injuries, indicating that ERS may play a vital role in the pathogenesis of LPS-induced hepatocyte injuries.

Key words: forensic pathology, endoplasmic reticulum, lipopolysaccharide, hepatocytes, apoptosis, rats

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