解剖学报

解剖学报 ›› 2013, Vol. 44 ›› Issue (3 ): 345-348.doi: 10.3969/j.issn.0529-1356.2013.03.010

• 组织学胚胎学发育生物学 • 上一篇    下一篇

Nrf2在急性亚砷酸钠暴露的小鼠胰岛β细胞中的表达

杨蓓*  陈雪 丁金兰   

  1. 中国医科大学基础医学院组织学与胚胎学教研室，沈阳 110001
  • 收稿日期:2012-06-08 修回日期:2013-01-06 出版日期:2013-06-06 发布日期:2013-07-16
  • 通讯作者: 杨蓓 E-mail:beibeiyang1978@gmail.com

Expression of Nrf2 induced by sodium arsenite in mouse pancreatic β cells

YANG Bei*  CHEN Xue  DING Jin-lan   

  1. Department of Histology and Embryology, College of Basic Medical Sciences, China Medical University, Shenyang 110001, China
  • Received:2012-06-08 Revised:2013-01-06 Online:2013-06-06 Published:2013-07-16

摘要：

目的 检测急性亚砷酸钠作用小鼠胰岛β细胞的Nrf2及其调控的II相解毒酶的表达情况。方法 4μmol/L亚砷酸钠（NaAsO2）作用于MIN6
细胞2、6、12、18、24h；利用Western blotting检测亚砷酸钠暴露不同时间点细胞核、细胞质和细胞总的Nrf2蛋白表达；应用Real-time PCR
检测不同时间点Nrf2及其调控的II相解毒酶NAD(P)H:醌氧化还原酶1（Nqo1）和血红素氧化酶1（Hmox-1）mRNA表达。结果 亚砷酸钠暴露2h，Nrf2
蛋白表达增多并达到高峰，之后随时间的延长Nrf2蛋白表达逐渐降低。Nrf2核蛋白表达的时间效应性与细胞总的Nrf2表达完全一致。但不同时间
点的亚砷酸钠暴露对Nrf2的mRNA表达未出现显著影响。Nqo1和Hmox-1表达亦存在时间效应性，即亚砷酸钠暴露后2h，Nqo1和Hmox-1 mRNA表达开
始增高，6h达到高峰，随后逐渐下降。结论 急性亚砷酸钠暴露后，能够诱导小鼠胰岛β细胞内Nrf2的蛋白表达，并促进其转位进入细胞核，调节
其下游II相解毒酶Nqo1和Hmox-1的转录活性。

关键词:  砷, 亚砷酸钠, 糖尿病, 胰岛β细胞, Nrf2, II相解毒酶, 免疫印迹法, 小鼠

Abstract:

Objective To test the expressions of Nrf2 and phase II detoxifying enzyme genes induced by sodium arsenite (NaAsO2)
in mouse pancreatic β-cell MIN6.Methods MIN6 cells were exposed to 4μmol/L NaAsO2 for 2, 6, 12, 18, 24hours. Nrf2 protein
levels in Nuclear factions, cytosolic factions and whole-cell of MIN6 cells were measured with Western blotting. mRNA expressions
of Nrf2 and phase II detoxifying enzyme genes were measured by Real-time PCR. Results Expression of Nrf2 protein was significantly
increased after 4μmol/L NaAsO2 exposure did reached the peak at 2hours, then decreased gradually. Nrf2 protein expressions in
nuclear factions were consistent with Nrf2 levels of the whole MIN6 cell in the time-dependent fasion. But 4μmol/L NaAsO2 exposure
did not induce a robust increase in Nrf2 mRNA. The induction of some phase II detoxification enzyme genes, including NAD(P)H:quinone
oxidoreductase 1 (Nqo1), heme oxygenase 1(Homx-1), were time-dependently increased in MIN6 cells. Nqo1 and Homx-1 mRNA expressions
were obviously increased after 2hours NaAsO2 exposure and went higher at 6hours, then decreased gradually. Conclusions Upon exposure
to NaAsO2, Nrf2 is stabilized and translocated into the nucleus, and then activates transcription of various detoxification
enzyme genes in mouse pancreatic β-cells.

Key words: Arsenic, Sodium arsenite, Diabetes, Pancreatic β-cell, Nrf2, Phase II detoxification enzyme, Western blotting, Mouse