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李平, 李芬, 叶昉, 吕玲, 陈军. Nrf 2信号通路在铅致SH-SY5Y细胞氧化应激中作用[J]. 中国公共卫生, 2012, 28(7): 933-935. DOI: 10.11847/zgggws-2012-28-07-18
引用本文: 李平, 李芬, 叶昉, 吕玲, 陈军. Nrf 2信号通路在铅致SH-SY5Y细胞氧化应激中作用[J]. 中国公共卫生, 2012, 28(7): 933-935. DOI: 10.11847/zgggws-2012-28-07-18
LI Ping, LI Fen, YE Fang, . Role of Nrf 2 signal pathway in lead-induced oxidative stress in SH-SY5Y cells[J]. Chinese Journal of Public Health, 2012, 28(7): 933-935. DOI: 10.11847/zgggws-2012-28-07-18
Citation: LI Ping, LI Fen, YE Fang, . Role of Nrf 2 signal pathway in lead-induced oxidative stress in SH-SY5Y cells[J]. Chinese Journal of Public Health, 2012, 28(7): 933-935. DOI: 10.11847/zgggws-2012-28-07-18

Nrf 2信号通路在铅致SH-SY5Y细胞氧化应激中作用

Role of Nrf 2 signal pathway in lead-induced oxidative stress in SH-SY5Y cells

  • 摘要: 目的 研究神经细胞内核因子E 2相关因子2(Nrf 2)信号通路是否对铅暴露所致的氧化应激产生应答及其可能机制。方法 用低、中、高剂量(5、25、125μmol/L)的醋酸铅溶液对人神经母细胞瘤SH-SY 5 Y细胞染毒,用2',7'-二氯荧光黄双乙酸盐(DCFH-DA)探针检测染毒2 h后细胞活性氧(ROS)水平,染毒24 h后用二硫基双硝基苯甲酸(DTNB)比色法检测还原性谷胱甘肽(GSH)水平,western blot法检测蛋白激酶C-δ(PKC-δ)、酪氨酸激酶2(CKⅡ)以及胞浆和胞核中Nrf 2蛋白的表达水平。结果 随着染毒剂量增加,低、中、高剂量组ROS含量依次为(559.17±54.56)、(585.50±36.41)、(621.00±29.96),与对照组(533.50±46.47)比较,中、高剂量组含量明显升高(P<0.05);GSH含量依次为(165.39±17.37)、(140.92±14.77)、(84.03±10.31),与对照组(222.10±14.91)比较,低、中、高剂量组含量均明显降低(P<0.01);与对照组比较,低、中、高剂量组胞浆Nrf 2相对灰度值为(0.38±0.09)、(0.27±0.09)、(0.25±0.11),胞浆Nrf 2表达均明显降低(P<0.05);低、中、高剂量组胞核Nrf 2相对灰度值为(1.38±0.50)、(1.55±0.49)、(2.79±1.56),仅高剂量组胞核Nrf 2蛋白表达明显增高(P<0.05)。低、中、高剂量组PKC-δ相对灰度值为(1.84±0.46)、(2.55±0.36)、(2.38±0.77),与对照组比较均明显升高(P<0.05);低、中、高剂量组CKⅡ相对灰度值为(1.28±0.32)、(1.34±0.21)、(1.52±0.42),与对照组比较仅高剂量组表达明显升高(P<0.05)。结论 铅致神经细胞氧化损伤的同时激活胞浆中Nrf 2转移入核内,进而发挥氧化应答,PKC-δ、CKⅡ在铅致Nrf 2激活过程中具有一定作用。

     

    Abstract: Objective To investigate the role of NF-E2-related factor 2(Nrf 2)signal pathway in lead-induced oxidative stress in SH-SY5Y cells as well as the possible mechanism. Methods SH-SY5Y cells were exposed to 0,5,25, and 125μmol/L lead acetate for 24 hours.After harvesting the cells,the level of reactive oxygen species(ROS)was measured by the method of 2',7'-dicholorofluorescin diacetate(DCFH-DA),and glutathione(GSH)was tested by dithiothymine double-nitrobenzonic acid(DTNB)method.Western blot was used to detect the levels of protein kinase C-theta(PKC-δ),casein kinase 2(CKⅡ)Nrf 2 in the cytoplasm and nucleus. Results Compared with the control group,the level of ROS of the moderate and high dose group were obviously increased(P<0.05),while the level of GSH in all groups obviously decreased(P<0.01).Compared with the control group,the protein expression level of Nrf 2 in the cytoplasm were significantly decreased(P<0.05).Meanwhile the protein expression levels of Nrf 2 in the nucleus of the high dose group showed a distinct elevatation,which was remarkbly different from that of the control group(P<0.05). Compared with the control group,the protein expression level of PKC-δ was significantly increased(P<0.05).The protein expression level of CKⅡof the high dose group showed a significant elevatation(P<0.05). Conclusion The findings demonstrate that lead can induce a nuclear accumulation of the transcription factor Nrf 2,which indicates the mediation of Nrf 2 in the cellular response against the oxidative stress caused by lead.The results also indicate that PKC-δ and CKⅡ play certain role in the Nrf 2 activation through increasing expression levels of two proteins stimulated by lead.

     

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