癌变·畸变·突变 ›› 2016, Vol. 28 ›› Issue (3): 169-173,178.doi: 10.3969/j.issn.1004-616x.2016.03.002

• 论著 • 上一篇    下一篇

亚硫酸钠对HL-7702细胞极低密度脂蛋白组装分泌和内质网应激的影响

张艳, 白剑英, 雷佩玉, 王幼萍, 赵春燕, 梁瑞峰   

  1. 山西医科大学公共卫生学院环境卫生学教研室, 山西 太原 030001
  • 收稿日期:2016-01-29 修回日期:2016-03-31 出版日期:2016-05-31 发布日期:2016-05-31
  • 通讯作者: 白剑英,E-mail:jybai66@aliyun.com E-mail:jybai66@aliyun.com
  • 作者简介:张艳,E-mail:yanxiayuxue@126.com。
  • 基金资助:
    国家自然科学基金(30972445);山西省回国留学人员科研资助项目(2014-034)

Effects of sodium sulfite on very low density lipoprotein assembly and secretion, and endoplasmic reticulum stress in HL-7702 liver cells

ZHANG Yan, BAI Jianying, LEI Peiyu, WANG Youping, ZHAO Chunyan, LIANG Ruifeng   

  1. Department of Environmental Health, School of Public Health, Shanxi Medical University, Taiyuan 030001, Shanxi, China
  • Received:2016-01-29 Revised:2016-03-31 Online:2016-05-31 Published:2016-05-31

摘要: 目的:研究亚硫酸钠(Na2SO3)是否通过抑制极低密度脂蛋白(VLDL)组装分泌和诱导内质网应激引起肝细胞脂肪变性。方法:分别经不同浓度(0.1、0.5、2.5、10 mmol/L)Na2SO3染毒人正常HL-7702肝细胞24和48 h后,采用酶联免疫吸附法(ELISA)和Western blot测定细胞外VLDL水平;采用荧光定量PCR法(qPCR)检测VLDL组装相关因子微粒体载脂蛋白B100(apoB100)、甘油三酯转移蛋白(MTP)、二脂酰甘油酰基转移酶2(DGAT2)、甘油三酯水解酶(TGH)的mRNA表达水平;采用Western blot法检测内质网应激相关重链结合蛋白(BiP)和钙连蛋白(Calnexin)的表达水平。结果:ELISA和Western blot结果显示染毒24 h后,与阴性对照组相比,各Na2SO3处理组VLDL含量均明显增加,差异均有统计学意义(P均<0.05);而作用48 h后,各Na2SO3处理组VLDL含量与阴性对照组相比均无明显变化(P均>0.05)。qPCR检测结果显示,与阴性对照组相比,染毒24 h,10 mmol/L Na2SO3组肝细胞内apoB100 mRNA表达升高(P<0.05);2.5和10 mmol/L Na2SO3可使肝细胞内TGH mRNA表达升高(P<0.05);染毒48 h,0.5和2.5 mmol/L Na2SO3引起肝细胞内TGH mRNA表达量明显升高(P<0.05)。Western blot结果显示染毒24 h和48 h后,不同浓度Na2SO3对细胞内BiP蛋白表达水平均无明显影响,但可以明显增加Calnexin蛋白表达(P<0.05)。结论:Na2SO3不能抑制VLDL组装分泌,但可能对肝细胞糖蛋白的正确折叠有一定干扰作用。

关键词: 亚硫酸钠, 极低密度脂蛋白组装, 内质网应激, 肝细胞

Abstract: OBJECTIVE:To investigate whether sodium sulfite (Na2SO3) induces lipid accumulation by inhibition of very low density lipoprotein (VLDL) assembly and activation of endoplasmic reticulum stress. METHODS:Human diploid liver cells (HL-7702) were cultured in DMEM medium containing 10% fetal bovine serum. Cells were exposed to different concentrations (0.1, 0.5, 2.5, 10 mmol/L) of Na2SO3 for 24 h and 48 h, and positive controls were used. VLDL secretion was detected by ELISA and Western blot, mRNA levels of VLDL assembly related factors were detected by quantitative real-time PCR (qPCR) and protein expressions of endoplasmic reticulum stress were detected by Western blot. RESULTS:ELISA and Western blot results showed that after exposure to Na2SO3 for 24 h, VLDL contents were significantly increased compared to the controls (P<0.05). After 48 hours, the difference became non-significant (P>0.05). QPCR test results showed that, compared with the negative control group, exposure to 10 mmol/L Na2SO3 for 24 h increased the expression of apoB100 mRNA significantly (P<0.05). In addition, the mRNA level of TGH in 10 mmol/L Na2SO3 and 2.5 mmol/L Na2SO3 groups were significantly increased (P<0.05). After exposure for 48 h, the mRNA levels of TGH in 2.5 mmol/L Na2SO3 and 0.5 mmol/L Na2SO3 groups were increased significantly (P<0.05). Protein expression of Calnexin was significantly increased (P<0.05) after Na2SO3 exposure. CONCLUSION:Na2SO3 may increase VLDL assembly and interfere with the correct folding of glycoprotein in hepatocytes.

Key words: sodium sulfite, very low density lipoprotein assembly, endoplasmic reticulum stress, hepatocytes

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