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周雪琴, 黄炜, 徐勇等. 高糖诱导的肾小球系膜细胞Smad4蛋白的SUMO修饰作用[J]. koko体育app 学报（医学版）, 2014, 45(3): 380-385.
引用本文: 周雪琴, 黄炜, 徐勇等. 高糖诱导型的肾小球系膜神经元Smad4淀粉酶的SUMO装饰用途[J]. 江西学校学报（医学专业版）, 2014, 45(3): 380-385.
ZHOU Xue-qin, HUANG Wei, XU Yong.et al. High Glucose Induces Sumoylation of Smad4 via SUMO2/3 in Glomerular Mesangial Cells[J]. Journal of Sichuan University (Medical Sciences), 2014, 45(3): 380-385.
Citation: ZHOU Xue-qin, HUANG Wei, XU Yong.et al. High Glucose Induces Sumoylation of Smad4 via SUMO2/3 in Glomerular Mesangial Cells[J]. Journal of Sichuan University (Medical Sciences), 2014, 45(3): 380-385.

高糖诱导的肾小球系膜细胞Smad4蛋白的SUMO修饰作用

High Glucose Induces Sumoylation of Smad4 via SUMO2/3 in Glomerular Mesangial Cells

  • 摘要: 目的 观察高糖刺激下大鼠肾小球系膜细胞小泛素相关修饰物 （SUMO）2/3和Smad4蛋白的表达及二者之间的相互作用，探讨SUMO化修饰在糖尿病肾病转化生长因子-β （TGF-β）信号通路中的作用与机制。 方法 将体外培养的大鼠肾小球系膜细胞分别设正常对照组 (5.6 mmol/L葡萄糖)，高糖组 （10、20、30 mmol/L葡萄糖），渗透压对照组 (5.6 mmol/L葡萄糖+24.4 mmol/L甘露醇）。用Western blot检测各组细胞SUMO2/3和Smad4蛋白表达；RT-PCR检测SUMO2/3和纤维连接蛋白 （fibronectin，FN） mRNA表达；免疫共沉淀方法检测SUMO2/3与Smad4蛋白间的相互作用；免疫荧光双染技术检测SUMO2/3与Smad4在细胞中的共定位关系。 结果 与正常对照组比较，各高糖组SUMO2/3、Smad4蛋白及SUMO2/3、FN mRNA表达均增加 （P<0.05）；免疫共沉淀结果显示SUMO2/3与Smad4在各组均存在相互作用，并在高糖刺激下显著增强 （P<0.05）；免疫荧光双染结果表明SUMO2/3与Smad4在肾小球系膜细胞内存在共定位现象,且在高糖组更明显。 结论 SUMO2/3可能通过共价修饰Smad4参与糖尿病肾病时TGF-β信号通路的调控。  
    Abstract: Objective To investigate the expression of SUMO2/3 and Smad4 in rat glomerular mesangial cells (GMCs) induced by high glucose, and the interaction between small ubiquitin related modifier （SUMO）2/3 and Smad4; and to explore the role and mechanism of sumoylation in regulating TGF-β signaling in diabetic nephropathy. Methods Cultured rat GMCs were divided into five groups: normal glucose group (5.6 mmol/L glucose), high glucose groups (10, 20 and 30 mmol/L glucose), and mannitol group (osmotic control). The expression of SUMO2/3, Smad4 and fibronectin (FN) was measured by Western blot and RT-PCR. The interaction and colocalization between SUMO2/3 and Smad4 were detected by Co-immunoprecipitation and immunofluorescence confocal laser microscopy. Results Compared with controls, the expressions of SUMO2/3, Smad4 and FN in the cells in the high glucose groups increased (P<0.05). The co-immunoprecipitation and immunofluorescence confocal laser scanning showed that Smad4 interacted and colocalized with SUMO2/3 and the sumolyation (SUMO2/3) of Smad4 was enhanced significantly in the high glucose groups compared with the control group (P<0.05). Conclusion Sumolyation of Samd4 by SUMO2/3 may be involved in the regulation of TGF-β signaling in diabetic nephropathy.  
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