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

作　　者：崔双杰[1] 巨晓芬 潘宝龙[1] 张玲[1] 路小婷[1] CUI Shuang-jie;JU Xiao-fen;PAN Bao-long;ZHANG Ling;LU Xiao-ting(Department of Occupational Health, School of Public Health, Shanxi Medical University, Taiyuan, Shanxi 030001, China）.)

机构地区：[1]山西医科大学公共卫生学院劳动卫生教研室,山西太原030001

出　　处：《环境与职业医学》2018年第4期361-365,共5页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金(编号:81001241;81472959)

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。[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 N2 a cells. [Methods] N2 a 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 N2 a 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 aluminumtrichloride 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 N2 a cells, which is possibly related to the expression changes of CHIP and Hsp70.

关 键 词：CHIP HSP70 铝 TAU蛋白 磷酸化 

分 类 号：R114[医药卫生—卫生毒理学]