醋酸铅诱导HK-2细胞凋亡及其机制研究  

Lead acetate-induced apoptosis in human renal proximal tubular epithelial cells(HK-2)and its mechanism

作　　者：金文达[1] 雷义[1] 朱茂祥[2] 陈锋[3] 曾燃元 

机构地区：[1]邵阳市疾病预防控制中心,湖南邵阳422000 [2]军事医学科学院放射与辐射医学研究所 [3]南华大学公共卫生学院环境医学研究所

出　　处：《毒理学杂志》2009年第5期364-367,共4页Journal of Toxicology

摘　　要：目的研究醋酸铅能否诱导人肾小管上皮细胞(HK-2)凋亡,并探讨其凋亡机制。方法平板克隆实验法检测细胞活力;AnnexinV(膜联蛋白V)-FITC(异硫氰酸荧光素)/PI(碘化丙啶)双染法结合流式细胞仪分析细胞凋亡率;荧光化学分光仪分析caspase-3-、8-、9(半胱天冬酶-3-、8-、9)的活性变化;Western bolt(免疫印迹法)检测bcl-2、bax蛋白的表达。结果铅可抑制HK-2细胞生长活力,随着铅浓度增高,细胞生长活力降低(P<0.05,P<0.01);铅使caspase-3、caspase-9的活性增强(P<0.05,P<0.01),但对caspase-8的活性无明显影响;caspase-3抑制剂可明显抑制铅诱导的caspase-3的活化,并抑制铅诱导的HK-2细胞凋亡;Western bolt结果显示铅染毒组HK-2细胞B细胞淋巴瘤/白血病-2(bcl-2)蛋白表达降低,bax蛋白表达增高。结论铅诱导HK-2细胞凋亡可能与铅增强caspase-3活性及抑制bcl-2蛋白表达有关。Objective To study the effect of lead acetate on the apoptosis of HK-2 cells and the apoptosis pathway.Methods The viabilities of cells were mediated by colony formation assay.The cell apoptosis was determined by flow cytometric analysis with Annexin-V（FITC） and Propidium Iodide（PI）.Activities of caspase-3,-8,-9 were determined by colorimetric assay.Western-bolting was utilized to measure the expressions ofbcl-2 and bax.Results Colony formation assay results revealed that lead could inhibit the viabilities of cells in does-dependent manner;The rates of apoptosis of lead-treated HK-2 cells increased significantly which were determined by Flow Cytometer with FITC-AnnexinVand PI double staining（P〈0.05,P〈0.01）;colorimetric assay results showed that lead could stimulated the activities of caspase-3,caspase-9,but had no effect on caspase-8 activity;Caspase-3 inhibitor,z-DEVD-CHO protected HK-2 from apoptosis induced by lead;Western-bolt result showed that lead stimulated the expression of bax,but inhibited bcl-2 expression.Conclusion The results were suggested that the apoptosis of HK-2 cells induced by lead might be associated with the caspase-3 activation、bcl-2 expression decreased and bax expressed increased.

关 键 词：铅 HK-2细胞 细胞凋亡 BCL-2 BAX 

分 类 号：R114[医药卫生—卫生毒理学]