Zhang Jun, Li Ping-hua, Yang Lei, DU Qing-sheng, Guo Ting-ting, Tang Xun
Department of Nephrology, Zhujiang Hospital, Southern Medical University, Guangzhou, China. E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2011 May;31(5):839-43.
To detect the effect of connective tissue growth factor (CTGF) on podocalyxin expression in mouse podocytes exposed to high glucose in vitro and explore the possible pathway involved.
The expression vector carrying a small interfering RNA (siRNA) targeting CTGF was transfected into mouse podocytes cultured in the presence of 1 g/L glucose (normal control), 4.5 g/L glucose (high glucose group), 1 g/L glucose + 3.5 g/L mannitol (iso-osmolar control group). The changes in the protein expression levels of podocalyxin, CTGF and ERK1/2 in the cells in response to the treatments were investigated using Western blotting.
High glucose exposure for 24 and 48 h resulted in significantly decreased expression of podocalyxin and increased CTGF in the podocytes (P<0.05). Phosphorylation of ERK1/2 occurred as early as 30 min after the exposure, and the activation was maintained till 24 h. Transfection of the cells with siRNA targeting CTGF significantly inhibited these changes.
CTGF is an important mediator of high glucose-induced podocyte damage and decreases the protein level of podocalyxin by the ERK1/2 pathway. CTGF-specific siRNA can alleviate high glucose-induced podocyte injury, suggesting its potential value in treatment of diabetic nephropathy.
检测结缔组织生长因子(CTGF)对体外高糖环境下小鼠足细胞中足萼蛋白表达的影响,并探讨其可能的作用途径。
将携带靶向CTGF的小干扰RNA(siRNA)的表达载体转染至在1 g/L葡萄糖(正常对照组)、4.5 g/L葡萄糖(高糖组)、1 g/L葡萄糖+3.5 g/L甘露醇(等渗对照组)条件下培养的小鼠足细胞中。采用蛋白质印迹法研究处理后细胞中足萼蛋白、CTGF和ERK1/2蛋白表达水平的变化。
高糖处理24小时和48小时后,足细胞中足萼蛋白表达显著降低,CTGF表达增加(P<0.05)。ERK1/2磷酸化在高糖处理后30分钟即出现,并持续激活至24小时。用靶向CTGF的siRNA转染细胞可显著抑制这些变化。
CTGF是高糖诱导足细胞损伤的重要介质,通过ERK1/2途径降低足萼蛋白水平。CTGF特异性siRNA可减轻高糖诱导的足细胞损伤,提示其在糖尿病肾病治疗中的潜在价值。