摘要
目的造影剂肾病的发病机制迄今尚未完全阐明,文中旨在观察碘普罗胺对人肾近曲小管上皮细胞(HK-2)细胞凋亡以及Klotho/Wnt/β-catenin信号通路的影响,探讨碘普罗胺引起急性肾损伤的可能机制。方法用不同浓度的碘普罗胺(37、74、111、148 mgI/mL)处理HK-2细胞24 h,应用CCK-8法测定细胞活力,DAPI染色法观察细胞核凋亡形态,Western blot法测定凋亡相关蛋白Bax、Bcl-2、Cleaved-caspase-3蛋白以及Klotho、Wnt、磷酸化-β-catenin(p-β-catenin)及β-catenin蛋白表达水平。另外,加入不同剂量的N-乙酰半胱氨酸(NAC)(2、4、8、16、32 mmol/L)处理,检测NAC对碘普罗胺诱导的HK-2细胞Klotho/Wnt/β-catenin信号通路相关蛋白表达的影响。结果与对照组相比,不同浓度的碘普罗胺处理组HK-2细胞活力明显降低(P <0.05);而细胞凋亡率明显增高(P <0.05);Cleaved-caspase-3、Wnt、p-β-catenin及β-catenin蛋白表达明显升高(P <0.05),而Klotho蛋白表达水平明显降低(P <0.05)。除了37mgI/mL碘普罗胺组外,其余碘普罗胺处理组Bax/Bcl-2比值均较对照组增高(P <0.05)。加入NAC处理后,能逆转碘普罗胺对Klotho/Wnt/β-catenin信号通路蛋白表达的影响(P <0.05)。结论碘普罗胺可诱导HK-2细胞发生凋亡,其机制可能与其下调Klotho蛋白表达,激活Wnt/β-catenin信号通路有关。
Objective The pathogenesis of contrast-induced nephropathy has not been fully elucidated. The purpose of this study was to observe the effect of iopromide on apoptosis and Klotho/Wnt/β-catenin signaling pathway in human proximal tubular cells(HK-2). Methods HK-2 cells were treated with different concentrations of iopromide(37,74,111,148 mgI/mL)for 24 h. The expression of apoptosis-related protein,such as Bax,Bcl-2 and Cleaved-caspase-3 and the expression of Klotho,Wnt,phospho-β-catenin(p-β-catenin)and β-catenin protein were detected by Western blot. In addition,different doses of N-acetylcysteine(NAC)(2,4,8,16,32 mmol/L) were added to detect the effects of NAC on Klotho/Wnt/β-catenin signaling pathway in HK-2 cells induced by iopro-mide. Results Compared with the control group,the viability of HK-2 cells treated with different concentrations of iopromide was significantly decreased,while the apoptosis rate was significantly increased;the protein level of Cleaved-caspase-3,Wnt,p-β-catenin,β-catenin were up-regulated,however,the expression of Klotho was down-regualted. In addition,the ratio of Bax/Bcl-2 was significantly increased except for the 37mgI/ml iopromide group (P<0.05). Furthermore,the effect of iopromide on Klotho/Wnt/β-catenin signaling pathway could be reversed by adding NAC(P<0.05). Conclusion Iopromide can induce apoptosis in HK-2 cells,the underlying mechanism may be related to down-regulation of Klotho protein expression and activation of Wnt/β-catenin signaling pathway.
作者
陈燕玲
罗婷
高昕乐
许志威
凡栋
吴秀香
靳俊峰
王智
赵晓朵
庄晓东
CHEN Yanling;LUO Ting;GAO Xinyue;XU Zhiwei;FAN Dong;WU Xiuxiang;JIN Junfeng;WANG Zhi;ZHAO Xiaoduo;ZHUANG Xiaodong(Department of Pathophysiology,Zhuhai Campus of Zunyi Medical University,Zhuhai 519041,China)
出处
《实用医学杂志》
CAS
北大核心
2019年第5期729-733,共5页
The Journal of Practical Medicine
基金
国家自然科学基金(编号:81560133)
贵州省科技合作计划项目(编号:黔科合LH字[2015]7529号)
合肥医学院博士启动基金(编号:F-711)
