摘要
目的探讨Th17细胞主要效应因子IL-17对足细胞特异性标记蛋白Nephrin和Podocalyxin及凋亡蛋白Caspase 8和Caspase 3的表达变化。方法分别以不同浓度(2,5,10,15ng/mL)重组IL-17刺激培养足细胞48h,采用实时荧光定量(Q-PCR)及Western blot在基因和蛋白水平分别检测足细胞标志性蛋白Nephrin和Podocalycin的表达,免疫荧光技术检测足细胞凋亡蛋白Caspase 8和Caspase 3的表达。结果不同浓度IL-17刺激足细胞,与对照组比较,足细胞特异性标记蛋白Nephrin和Podocalycin的mRNA和蛋白水平表达降低(P<0.05),足细胞凋亡蛋白Caspase 3和Caspase 8表达升高(P<0.05),不同浓度刺激组间无显著性差异。结论重组IL-17体外刺激培养足细胞导致足细胞损伤及凋亡,此研究结果提示或可在肾小球疾病患者中通过采用IL-17抗体或IL-17siRNA抑制内源性IL-17的表达从而抑制足细胞的损伤及凋亡,从而降低肾小球疾病蛋白尿的产生,为深入研究肾脏疾病的病因和发病机制提供突破口,进而找到特异有效的治疗,有重要的基础和临床研究价值。
Objective To investigate the effects of IL-17 on significance of Nephrin and Podocylaxin in podocyte,To investigate the injury and apoptosis of podocyte induced by IL-17.Methods Different concentrations of IL-17 (2,5,10,15 ng/mL)were stimulated Podocytes for 48 hours,The mRNA of nephrin and Podocylaxin were quantified by Q-PCR while the level of protein were validated by Western blotting.Further,Caspase 8 and Caspase 3 were visualized by immunofluorescence analysis.Results Compared to the control group,the mRNA and protein of nephrin and Podocylaxin showed a lower expression after stimulating with IL-17 (P〈0.05),however,the Caspase 8 and Caspase 3 revealed a opposite expression (P〈0.05).Conclusion IL-17 stimulation causes the podocyte injury and apoptosis,which shows that inhibiting the expression of IL-17 via IL-17 antibodies or IL-17 siRNA may avoid podocyte injury and apoptotic in Patients with glomerular diseases,which lay foundation on reducing proteinuria and glomerular disease for further research on the etiology and pathogenesis of kidney disease,it may provide a specific effective treatment,and also has significant value of clinical research.
出处
《贵州医药》
CAS
2017年第2期115-117,共3页
Guizhou Medical Journal
基金
国家自然科学基金项目(项目编号:81460138)