XIAP蛋白表达改变在铅诱导小鼠海马神经元细胞损伤中的作用

Role of XIAP expression change in lead- induced mouse hippocampal neuron HT22 cell injury

目的探讨重金属铅暴露诱导小鼠海马神经元HT22细胞X连锁凋亡抑制蛋白（X1AP）表达的变化及其与小鼠海马神经元细胞损伤程度的关系。方法建立小鼠海马神经元细胞铅暴露模型,采用噻唑蓝法（MTT）筛选HT22细胞的最佳生长密度,并检测醋酸铅0～100μM暴露24 h和48 h后细胞活力的变化。Western blot方法检测HT22细胞铅暴露后XIAP蛋白表达水平的变化,免疫细胞荧光染色方法检测HT22细胞铅暴露后XIAP蛋白的表达水平及分布变化。结果 MTT法检测结果显示HT22细胞在5和10μM醋酸铅处理48 h后,细胞增殖活力均明显低于对照组（5μM,P〈0.05;10μM,P〈0.01）。Western blot表明5和10μM醋酸铅处理24和48 h组HT22细胞的XIAP蛋白表达水平低于对照组,差异有统计学意义（P〈0.05）。免疫组织化学染色显示醋酸铅处理48 h后,HT22细胞中的XIAP蛋白也明显低于对照组。结论 XIAP蛋白可能参与了铅暴露导致小鼠海马神经元细胞损伤。

Objective To investigate the role of X- linked inhibitor of apoptosis protein（ XIAP） expression in lead induced mouse hippocampal neuron HT22 cell injury. Methods We established lead exposure cell model in mouse HT22 cells. Methyl thiazolyl tetrazolium（ MTT） assay was performed to determine the best proliferation density of HT22 cells and viability alterations of HT22 cells exposed to lead acetate at 0 ～ 100μM for 24 h and 48 h. The expression levels of XIAP in HT22 cells after lead exposure were detected by Western blot,and the expression and distribution of XIAP in HT22 cells were determined by immunofluorescence staining. Results MTT assay showed that cell proliferation was inhibited in HT22 cells by exposure to 5 and 10μM lead acetate for 48h（ P 〈0. 05 and P 〈0. 01,respectively）. Western blot showed that XIAP expression decreased in HT22 cells treated with 5 and 10μM lead acetate when compared with the controls（ P〈 0. 05）. Immunofluorescence staining showed that lead acetate exposure for 48 h could significantly reduced the XIAP expression in HT22 cells. Conclusions XIAP may be involved in the lead- induced mouse hippocampus neuron HT22 cell injury.