Li Zhuo, Zhang Li, Shi Wei, Chen Yuanhan, Zhang Hong, Liu Shuangxin, Liang Xinling, Ling Ting, Yu Chunping, Huang Zhongshun, Tan Xiaofan, Zhao Xinchen, Ye Zhiming, Zhang Bin, Wang Wenjian, Li Ruizhao, Ma Jianchao
Graduate School, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.
Department of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510080, P.R. China.
Mol Med Rep. 2015 Nov;12(5):6849-54. doi: 10.3892/mmr.2015.4295. Epub 2015 Sep 7.
Integrin β1 and β3 expression by podocytes is required to maintain glomerular structural integrity. Previous studies have shown that aldosterone (ALD) is involved in glomerular podocyte injury, and mineralocorticoid receptor (MR) blocker spironolactone effectively reduces proteinuria in patients with diabetic nephropathy. The present study was designed to observe the effects of spironolactone on β1 and β3 integrin expression and podocyte motility under in vitro diabetic conditions. Immortalized mouse podocytes were cultured in media containing normal glucose (NG) levels, high glucose (HG) or HG plus spironolacton. The expression of β1 and β3 integrin in podocytes was detected by reverse transcription quantitative polymerase chain reaction, immunofluorescence and western blot analyses. The effects of spironolacton on podocyte motility was further evaluated using a wound healing assay. HG stimulation markedly decreased mRNA and protein expression of integrin β1, and significantly increased mRNA and protein expression of integrin β3 in cultured podocytes. However, simultaneous treatment with spironolacton (10‑7 mol/l) significantly attenuated HG-mediated increases in integrin β3 and decreases in integrin β1 expression. Furthermore, the migration of podocytes induced by HG was abrogated by concomitant treatment with spironolacton. In conclusion, the present study suggested that HG decreased the expression of integrin β1 in cultured podocytes, accompanied with an increase of integrin β3. Spironolactone inhibited cell motility and stabilized podoctyes treated with HG, probably through partly normalizing the expression of integrin β1 and decreasing the expression of integrin β3.
足细胞表达整合素β1和β3是维持肾小球结构完整性所必需的。先前的研究表明，醛固酮（ALD）参与肾小球足细胞损伤，而盐皮质激素受体（MR）阻滞剂螺内酯可有效降低糖尿病肾病患者的蛋白尿。本研究旨在观察螺内酯在体外糖尿病条件下对β1和β3整合素表达及足细胞运动性的影响。将永生化小鼠足细胞培养于含有正常葡萄糖（NG）水平、高葡萄糖（HG）或HG加螺内酯的培养基中。通过逆转录定量聚合酶链反应、免疫荧光和蛋白质印迹分析检测足细胞中β1和β3整合素的表达。使用伤口愈合试验进一步评估螺内酯对足细胞运动性的影响。HG刺激显著降低培养足细胞中整合素β1的mRNA和蛋白质表达，并显著增加整合素β3的mRNA和蛋白质表达。然而，同时用螺内酯（10-7 mol/l）处理可显著减弱HG介导的整合素β3增加和整合素β1表达降低。此外，HG诱导的足细胞迁移被螺内酯联合处理所消除。总之，本研究表明HG降低了培养足细胞中整合素β1的表达，同时伴有整合素β3的增加。螺内酯可能通过部分使整合素β1表达正常化并降低整合素β3的表达来抑制细胞运动性并稳定经HG处理的足细胞。
