瞬时受体电位阳离子通道蛋白6和整合素连接激酶在以蛋白尿为主要表现的慢性肾脏病中的表达  

Expressions of transient receptor potential cation channel 6 and integrin- linked kinase in glomerular of proteinuric kidney diseases

作　　者：郭晓红[1] 张涵[1] 胡杰[1] 谭小猛 杨洁[1] 孙脊峰[1] 

机构地区：[1]第四军医大学唐都医院肾脏内科,西安710038

出　　处：《中华肾脏病杂志》2017年第5期378-384,共7页Chinese Journal of Nephrology

基　　金：国家自然科学基金（81170685）;陕西省社会发展科技攻关项目（2011K14-09-05）

摘　　要：目的观察以蛋白尿为主要表现的肾脏病患者的肾小球内瞬时受体电位阳离子通道蛋白6（TRPC6）和整合素连接激酶（ILK）的表达和分布，并通过体外实验探讨过表达TRPC6对ILK的影响。方法选取2012年到2013年于本院进行肾穿刺活检术且24h尿蛋白量大于1g的患者，通过免疫组织化学法观察TRPC6和ILK在肾活检组织中肾小球内的表达和分布。体外培养MPC5足细胞，以10^-7mol／L阿霉素（ADR）刺激足细胞12、24、36h。采用脂质体2000试剂将pcDNA3．1（＋）-TRPC6质粒、pcDNA3．1（＋）转染入MPC5足细胞分别建立TRPC6过表达组和阴性对照组。Western印迹检测TRPC6和ILK蛋白的表达。结果纳入膜性肾病14例，局灶节段性肾小球硬化症（FSGS）13例，膜增生性肾小球肾炎15例，系膜增生性肾小球肾炎12例，增生硬化性肾小球肾炎10例，IgA肾病15例，紫癜性肾炎13例，狼疮肾炎15例，高血压肾损伤13例和糖尿病肾病14例，及正常肾组织9例。TRPC6主要表达于足细胞，除高血压肾损伤外，其余肾组织TRPC6表达评分均高于正常肾组织（均P〈0．05）；ILK表达于足细胞与系膜区，FSGS、狼疮肾炎及糖尿病肾病的肾组织中ILK表达得分均高于正常。肾组织（均P〈0．05），其他肾病组织ILK表达高于正常肾组织，但差异均无统计学意义（均P〉0．05）；FSGS和糖尿病。肾病肾组织中TRPC6和ILK的表达呈正相关（分别r=0．906，P〈0．001；／-=0．783，P=0．001）。体外ADR刺激24h和36h时TRPC6和ILK蛋白的表达均高于正常对照组（均P〈0．05）；TRPC6过表达组足细胞ILK蛋白表达与阴性对照组和正常对照组比较均增加（均P〈0．05）。结论以蛋白尿为主要表现的。肾脏病患者的肾小球内TRPC6和ILK表达升高。尤其是FSGS和糖尿病肾病。上调足细胞TRPC6表达可增加ILK蛋白表达，两者可能共同参与足细胞损伤。Objective To observe the expressions and distribution of transient receptor potential cation channel 6 （TRPC6） and integrin-linked kinase （ILK） in the glomeruli of renal biopsy tissue of patients with proteinuric kidney diseases, and to investigate the effect of TRPC6 over- expression on ILK in vitro. Methods The archival histological specimens of patients admitted to Tangdu hospital from 2012 to 2013, with 24- hour urinary protein over 1 g, were collected. The expressions and distribution of TRPC6 and ILK in the glomeruli of renal biopsy tissue were observed by immunohistochemistry. MPC5 podocytes were cultured in vitro and they were stimulated with 10^-7 mol/L ADR for 12, 24 and 36 h. The pcDNA3.1（＋）-TRPC6 plasmid and pcDNA3.1（＋） were transfected into MPC5 podocytes by liposome 2000 reagent to establish the TRPC6 overexpression group and the negative control group respectively. Western blotting was used to detect the expressions of TRPC6 and ILK protein. Results There were 14 cases of membranous nephropathy, 13 cases of focal segmental glomerulosclerosis （FSGS）, 15 cases of membranoproliferative glomerulonephritis, 12 cases of mesangial proliferative glomerulonephritis, 10 cases of hyperplastic sclerosis nephritis, 15 cases of IgA nephropathy, 13 cases of purpura nephritis, 15 cases of lupus nephritis, 13 cases of hypertensive renal injury, 14 cases of diabetic nephropathy and 9 cases of normal renal tissue included. In glomerulus, TRPC6 was expressed mainly in podocytes, and the expressions of TRPC6 in these renal tissues were higher than that in normal renal tissues （all P 〈 0.05）, except for hypertensive nephropathy. ILK was expressed in podoeytes and the mesangial areas. The expressions of ILK in FSGS, lupus nephritis and diabetic nephropathy were higher than that in normal kidney tissue （all P 〈 0.05）, while the other renal tissues was high but showed no statistical difference with normal kidney tissue （all P 〉 0.05）. The expressions of TRPC6 and ILK were positivel

关 键 词：肾功能不全 慢性 足细胞 TRPC阳离子通道 蛋白质丝氨酸苏氨酸激酶 

分 类 号：R692[医药卫生—泌尿科学]