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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 investigate whether lead acetate can induce apoptosis of human renal tubular epithelial cells (HK-2) and to explore its mechanism of apoptosis. Methods Cell viability was assayed by plate clone assay; apoptosis rate was analyzed by Annexin V (Annexin V) -FITC / PI (propidium iodide) staining combined with flow cytometry; The activity of caspase-3-, 8-, 9 (caspase-3, 8-, 9) was analyzed by spectrophotometer. The expression of bcl-2 and bax protein was detected by Western blot. Results Lead inhibited the viability of HK-2 cells. With the increase of lead concentration, the viability of cells was decreased (P <0.05, P <0.01), and the activities of caspase-3 and caspase- 0.01), but no significant effect on the activity of caspase-8; caspase-3 inhibitor can significantly inhibit lead-induced caspase-3 activation and inhibit lead-induced apoptosis of HK-2 cells; Group B HK-2 cell B-cell lymphoma / leukemia-2 (bcl-2) protein expression decreased, bax protein expression increased. Conclusion Lead-induced apoptosis in HK-2 cells may be related to lead-induced caspase-3 activation and suppression of bcl-2 protein expression.