氯化钴诱导低氧对裸鼹鼠肝星形细胞增殖及凋亡的影响

doi:10.3969/j.issn.1674-5817.2016.01.014

实验动物与比较医学 ›› 2016, Vol. 36 ›› Issue (1): 66-71.DOI: 10.3969/j.issn.1674-5817.2016.01.014

氯化钴诱导低氧对裸鼹鼠肝星形细胞增殖及凋亡的影响

肖邦¹, 李莉², 余琛琳¹, 赵善民¹, 林丽芳¹, 杨文静¹, 丛薇¹, 崔淑芳¹

1.第二军医大学实验动物中心, 上海200433;
2.第二军医大学教学保障处, 上海200433

收稿日期:2015-12-14 出版日期:2016-02-25 发布日期:2016-02-25
作者简介:肖邦, 男(1989-), 硕士生。E-mail: xiaobangjinli@163.com
基金资助:
国家科技支撑计划课题(2015BAI09B02)、国家自然科学基金(31402028)、上海市科委基金 (14140900200, 13140900400)、全军实验动物专项课题[SYDW(2014)007]联合资助

Influence of Cobalt Chloride Induced Hypoxia on Proliferation and Apoptosis of Hepatic Stellate Cells in Naked Mole Rat

XIAO Bang¹, LI Li², YU Chen-lin, ZHAO Shan-min¹, LIN Li-fang¹, YANG Wen-jing¹, CONG Wei¹, CUI Shu-fang¹

1. Laboratory Animal Center;
2. Teaching Guarantee Department, Second Military University, Shanghai 200433, China

Received:2015-12-14 Online:2016-02-25 Published:2016-02-25

摘要/Abstract

摘要： 目的比较研究氯化钴(CoCl₂)对裸鼹鼠及C57BL/6J小鼠肝星形细胞增殖和凋亡的影响。方法采用活细胞计数试剂盒(CCK8)法和流式细胞术分别检测不同浓度CoCl₂对裸鼹鼠和小鼠肝星形细胞增殖活力以及和凋亡水平的影响。采用蛋白印迹测定CoCl₂处理后裸鼹鼠肝星形细胞低氧诱导因子1〈(HIF-1〈)和凋亡相关蛋白(BCL₂、BAX)的表达水平。结果低浓度(50 μmol/L)CoCl₂对C57BL/6j小鼠肝星形细胞增殖活性具有明显的抑制作用,并能显著提高其凋亡率,然而同样剂量的CoCl₂能够显著提高裸鼹鼠肝星形细胞的增值活性,对细胞凋亡率亦无明显影响。高剂量CoCl₂ (>200 μmol/L)虽能导致裸鼹鼠肝星形细胞增殖活力的降低,及凋亡率的升高,但裸鼹鼠肝星形细胞的变化幅度明显小于小鼠。同时CoCl₂处理后,裸鼹鼠肝星形细胞HIF-1〈的表达或累积显著升高(P<0.05),BCL₂/BAX比率显著上调(P<0.05)。结论与C57BL/6J小鼠相比,裸鼹鼠肝星形细胞具有更强的抵抗CoCl₂引起的化学性低氧损伤的能力。同时HIF-1〈可能参与调控裸鼹鼠抵抗化学性低氧环境造成的损伤。

关键词: 氯化钴(CoCl₂), 裸鼹鼠, 肝星形细胞, 凋亡

Abstract: Objective To compare and study the influence of cobalt chloride (CoCl₂) on proliferation and apoptosis of hepatic stellate cells in the naked mole rat and C57BL/6J mouse, and preliminarily investigate the possible mechanism of hypoxic tolerance in naked mole rat. Methods Evaluating the effects of different concentrations of CoCl₂ on proliferation activity and apoptosis level of hepatic stellate cells in the naked mole rat and mouse by Cell Counting Kit-8 assay (CCK8) and flow cytometry. Detect the expression of hypoxia inducible factor 1〈 (HIF-1〈) and apoptosis-related proteins (BCL₂, BAX)of hepatic stellate cells in the naked mole rat after CoCl₂ treatment by Western blot assay. Results CoCl₂significantly inhibited the proliferation and increased the apoptotic rate of hepatic stellate cells in C57BL/6j mice at a low concentration of 50 μmol/L. However, a same dose of CoCl₂ can significantly improve the proliferation of hepatic stellate cells in the naked mole rat but had no significant effect on its apoptotic rate. While high concentrations of CoCl₂ (> 200 μmol/L) showed a inhibition of proliferation and a rise of apoptotic rate of hepatic stellate cells in naked mole rat and mice, but the CoCl₂-induced changes in hepatic stellate cells of the naked mole rat were much smaller than those of mice. Simultaneously, the expression of HIF-1〈 and the ratio of apoptosis-related proteins (BCL₂/BAX) in hepatic stellate cells of the naked mole rat upregulated or accumulated significantly after treating with CoCl₂ (P<0.05). Conclusion Compared with C57BL/6J mice, the naked mole rat hepatic stellate cells had a greater ability of resistance to hypoxic damage caused by CoCl₂. It was speculated that HIF-1〈 may be involved in the regulation of resistance to damage caused by hypoxic environment in naked mole rat.

Key words: Cobalt chloride(CoCl₂), Naked mole rat, Hepatic stellate cells, Apoptosis

中图分类号:

Q95-33

肖邦, 李莉, 余琛琳, 赵善民, 林丽芳, 杨文静, 丛薇, 崔淑芳. 氯化钴诱导低氧对裸鼹鼠肝星形细胞增殖及凋亡的影响[J]. 实验动物与比较医学, 2016, 36(1): 66-71.

XIAO Bang, LI Li, YU Chen-lin, ZHAO Shan-min, LIN Li-fang, YANG Wen-jing, CONG Wei, CUI Shu-fang. Influence of Cobalt Chloride Induced Hypoxia on Proliferation and Apoptosis of Hepatic Stellate Cells in Naked Mole Rat[J]. Laboratory Animal and Comparative Medicine, 2016, 36(1): 66-71.

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氯化钴诱导低氧对裸鼹鼠肝星形细胞增殖及凋亡的影响

Influence of Cobalt Chloride Induced Hypoxia on Proliferation and Apoptosis of Hepatic Stellate Cells in Naked Mole Rat

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