摘要
目的探讨丙酮醛对人肾近端小管上皮细胞(HK-2细胞)损伤的作用及可能机制。方法采用不同浓度的丙酮醛(100、200、400、800、1 600μmol/L)处理HK-2细胞24 h后,应用CCK-8法检测HK-2细胞的存活率;罗丹明123染色法检测细胞线粒体膜电位;DAPI染色法观察HK-2细胞凋亡形态学变化。Western blot法检测HK-2细胞内磷酸化ERK1/2(phosphated ERK1/2,p-ERK1/2)、Bax及cleaved-caspase-3蛋白表达水平。结果与对照组相比,不同的浓度的丙酮醛均能降低HK-2细胞存活率(P<0.01);且HK-2细胞经丙酮醛诱导后cleaved-caspase-3(P<0.01)、Bax及p-ERK1/2蛋白表达水平均明显增高(P<0.05)。结论丙酮醛能诱导HK-2细胞出现明显的损伤及凋亡,其机制可能与其激活ERK1/2信号通路有关。
Objective To investigate the effects of different concentrations of methylglyoxal on the injury of hu- man tubule epithelial cells （ HK - 2 cells） and its mechanisms. Methods HK - 2 cells were incubated with dif- ferent concentrations of methylglyoxal （ 100,200,400,800 and 1 600 μmol/L） for 24 h. Cell viability was evalua- ted by CCK - 8 assay. Mitochondrial membrane potential was detected by rhodamine - 123. The morphological changes in apoptotic cells were measured by DAPI. The expression of cleaved caspase - 3, Bax and p - ERK was detected by western blot assay. Results CCk -8 assay showed that methylglyoxal significantly decreased the viability of HK - 2 cells in a dose - dependent manner （ P 〈 0.01 ）. Also, methylglyoxal reduced mitochondrial membrane potential in HK - 2 cells. In addition, the expressions of cleaved caspase - 3, Bax and p - ERK1/2 were up - regulated by methylglyoxal （P 〈 0.05 ）. Conclusion Methylglyoxal induces significant injury and apoptosis in HK -2 cells. The underlying mechanism might be involved in the activation of ERK1/2 signaling pathway.
出处
《遵义医学院学报》
2017年第5期526-530,535,共6页
Journal of Zunyi Medical University
基金
国家自然科学基金资助项目(NO:81560133)
贵州省科技合作计划项目(NO:黔科合LH字[2015]7529)
遵义医学院博士科研启动基金(NO:F-711)
作者简介
[通信作者]庄晓东，男，博士，研究方向：肾脏疾病的发病机制及药物防治，E-mail：kangfuyanling@126．com。
