崔双杰, 巨晓芬, 潘宝龙, 张玲, 路小婷. CHIP和Hsp70在三氯化铝致N2a细胞tau蛋白异常磷酸化中的作用[J]. 环境与职业医学, 2018, 35(4): 361-365. DOI: 10.13213/j.cnki.jeom.2018.17645
引用本文: 崔双杰, 巨晓芬, 潘宝龙, 张玲, 路小婷. CHIP和Hsp70在三氯化铝致N2a细胞tau蛋白异常磷酸化中的作用[J]. 环境与职业医学, 2018, 35(4): 361-365. DOI: 10.13213/j.cnki.jeom.2018.17645
CUI Shuang-jie, JU Xiao-fen, PAN Bao-long, ZHANG Ling, LU Xiao-ting. Effects of CHIP and Hsp70 on abnormal phosphorylation of tau protein induced by aluminum trichloride in N2a cells[J]. Journal of Environmental and Occupational Medicine, 2018, 35(4): 361-365. DOI: 10.13213/j.cnki.jeom.2018.17645
Citation: CUI Shuang-jie, JU Xiao-fen, PAN Bao-long, ZHANG Ling, LU Xiao-ting. Effects of CHIP and Hsp70 on abnormal phosphorylation of tau protein induced by aluminum trichloride in N2a cells[J]. Journal of Environmental and Occupational Medicine, 2018, 35(4): 361-365. DOI: 10.13213/j.cnki.jeom.2018.17645

CHIP和Hsp70在三氯化铝致N2a细胞tau蛋白异常磷酸化中的作用

Effects of CHIP and Hsp70 on abnormal phosphorylation of tau protein induced by aluminum trichloride in N2a cells

  • 摘要: 目的 探究热休克蛋白70羧基末端结合蛋白(CHIP)和热休克蛋白70(Hsp70)在三氯化铝致小鼠神经母细胞瘤细胞(N2a细胞)tau蛋白异常磷酸化中的作用。

    方法 以低、中、高剂量三氯化铝溶液(铝离子浓度分别为0.5、1.0、2.0 mmol/L)染毒N2a细胞,以不含三氯化铝溶液染毒者作为对照组,染毒48 h后显微镜下观察细胞形态,CCK-8法检测细胞活力,用Western blot法测定细胞tau-5蛋白、磷酸化蛋白(pThr181、pThr231、pSer262、pSer396)以及CHIP和Hsp70蛋白表达情况。

    结果 随着染毒剂量增加,细胞数量逐渐减少,突触回缩,胞体变圆。中、高剂量组细胞活力百分比(91.37±0.03)%和(78.45±0.10)%明显低于对照组(100.00±0.00)%(P<0.05,P<0.01)。高剂量组tau-5蛋白表达水平明显高于对照组(P<0.01);中、高剂量组pThr231蛋白表达水平明显高于对照组(P<0.01);各染毒组pSer396蛋白表达水平明显高于对照组(P<0.05或P<0.01);各组pThr181、pSer262蛋白表达水平的差异无统计学意义(P>0.05)。各染毒组CHIP、Hsp70表达水平均高于对照组(P<0.05或P<0.01)。

    结论 三氯化铝可以导致N2a细胞pThr231、pSer396表达增高,其作用机制可能与CHIP和Hsp70表达变化有关。

     

    Abstract: Objective To investigate the effects of carboxyl terminus of Hsc70-interacting protein (CHIP) and hot shock protein 70 (Hsp70) on aluminum trichloride induced abnormal phosphorylation of tau protein in mouse neuroblastoma N2a cells.

    Methods N2a cells were cultured with low, middle, and high doses of aluminium trichloride (0.5, 1.0, and 2.0 mmol/L, respectively), and the control group was treated without aluminium trichloride. The cell morphological changes were observed after 48 h of exposure under microscope and the cell viability was measured by CCK-8 assay. The protein expression levels of tau-5, pThr181, pThr231, pSer262, pSer396, CHIP, and Hsp70 were measured by Western blot.

    Results As the aluminum trichloride concentration increased, the count of N2a cells decreased, the synapses retracted, and the cell body rounded. The middle and high dose groups showed significantly lower cell viabilities(91.37±0.03)% and (78.45±0.10)% than the control group(100.00±0.00)% (P < 0.05, P < 0.01). The expression level of tau-5 protein of the high dose group was significantly higher than that of the control group (P < 0.01); the expression levels of pThr231 of the middle and high dose groups were significantly higher than that of the control group (P < 0.01); the expression levels of pSer396 were higher in all aluminum trichloride groups than in the control group (P < 0.05 or P < 0.01); but no differences were found in the expression levels of pThr181 and pSer262 (P > 0.05). The expression levels of CHIP and Hsp70 of all aluminium trichloride groups were significantly higher than those of the control group (P < 0.05 or P < 0.01).

    Conclusion Aluminum trichloride could increase the expression levels of pThr231 and pSer396 in N2a cells, which is possibly related to the expression changes of CHIP and Hsp70.

     

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