纤溶酶原激活物抑制因子1在腹膜透析患者中的表达及其对腹膜纤维化的影响  

Plasminogen activator inhibitor 1 expression in peritoneal dialysis patients and its effect on peritoneal fibrosis

作　　者：苏瑞 梅春丽 李玉萍 李铭 李婷[3] 赵欣 卢家美[4] 邹晓荣 SU Rui;MEI Chunli;LI Yuping;LI Ming;LI Ting;ZHAO Xin;LU Jiamei;ZOU Xiaorong(Department of Nephrology,986 th Hospital of Chinese People’s Liberation Army Air Force,Xi’an 710054,China;Department of Endocrinology,986 th Hospital of Chinese People’s Liberation Army Air Force;Department of Special Diagnosis,Shaanxi Provincial People’s Hospital;Department of Nephrology,Second Affiliated Hospital of Xi’an Jiaotong University)

机构地区：[1]中国人民解放军空军第九八六医院肾脏病科,西安710054 [2]中国人民解放军空军第九八六医院内分泌科 [3]陕西省人民医院特诊科 [4]西安交通大学第二附属医院肾病内科

出　　处：《山西医科大学学报》2021年第9期1187-1194,共8页Journal of Shanxi Medical University

基　　金：国家自然科学基金青年科学基金项目(81700605)。

摘　　要：目的探究纤溶酶原激活物抑制因子1(PAI-1)在腹膜透析患者中的表达及其功能。方法通过免疫组化和Wes-tern blot检测30例腹膜透析患者和30例终末期肾病患者腹膜组织中PAI-1的表达水平。将人腹膜间皮细胞(HPMCs)分为3组:正常对照组、TGF-β1+si-NC组和TGF-β1+si-PAI-1组。TGF-β1+si-NC组和TGF-β1+si-PAI-1组细胞用10 ng/ml TGF-β1培养24 h,并分别转染靶向PAI-1的siRNA(si-PAI-1)和阴性对照(si-NC)。通过Western blot检测腹膜间皮细胞中PAI-1、p-STAT3、STAT3、p-Smad3、Smad3、CollagenⅠ、E-cadherin、α-SMA、EGFR、p-EGFR和Acetylα-tubulin的蛋白表达。对C57/BL6小鼠连续28 d腹腔注射3 ml含4.25%葡萄糖的腹膜透析液来建立腹膜纤维化(PF)模型。将小鼠随机分为3组,每组12只:正常对照组、PF+si-NC组和PF+si-PAI-1组。PF+si-NC组和PF+si-PAI-1组小鼠分别腹腔注射300μl si-NC或si-PAI-1(0.1 mg/kg)。通过免疫组织化学染色检测小鼠腹膜组织中PAI-1、CollagenⅠ和α-SMA的阳性表达。结果与终末期肾病组比较,腹膜透析组患者腹膜组织中的PAI-1表达水平显著升高(P<0.05)。与正常对照组比较,TGF-β1+si-NC组腹膜间皮细胞中PAI-1、CollagenⅠ、α-SMA、p-Smad3、p-EGFR和p-STAT3蛋白相对表达量显著升高(P<0.05),E-cadherin、Acetylα-tubulin显著降低(P<0.05)。与TGF-β1+si-NC组比较,TGF-β1+si-PAI-1组腹膜间皮细胞中PAI-1、CollagenⅠ、α-SMA、p-Smad3、p-EGFR和p-STAT3蛋白相对表达量显著降低(P<0.05),E-cadherin、Acetylα-tubulin显著升高(P<0.05)。与正常对照组比较,PE+si-NC组腹膜组织中E-cadherin阳性率显著降低,α-SMA显著增加(P<0.05)。与PE+si-NC组比较,PE+si-PAI-1组腹膜组织中E-cadherin阳性率显著增加,α-SMA显著降低(P<0.05)。结论本研究证明了下调PAI-1可抑制腹膜纤维化的发展,推测这种抗纤维化机制可能与TGF-β1/Smad3和EGFR/STAT3信号通路的失活有关。Objective To explore the expression and function of plasminogen activator inhibitor 1(PAI-1)in peritoneal dialysis patients.Methods The expression of PAI-1 in the peritoneal tissues of 30 peritoneal dialysis patients and 30 end-stage renal disease patients was detected by immunohistochemistry and Western blot.Human peritoneal mesothelial cells(HPMCs)were divided into three groups:normal control group,TGF-β1+si-NC group and TGF-β1+si-PAI-1 group.HPMCs in TGF-β1+si-NC group and TGF-β1+si-PAI-1 group were cultured with 10 ng/ml TGF-β1 for 24 h and transfected with siRNA targeting PAI-1(si-PAI-1)and negative control(si-NC),respectively.The protein expression levels of PAI-1,p-STAT3,STAT3,p-Smad3,Smad3,CollagenⅠ,E-cadhe-rin,α-SMA,EGFR,p-EGFR and Acetylα-tubulin in peritoneal mesothelial cells were detected by Western blot.C57/BL6 mice were intraperitoneally injected with 3 ml of peritoneal dialysate containing 4.25%glucose for 28 consecutive days to establish a peritoneal fibrosis(PF)model.The mice were randomly divided into three groups(each with 12 mice):normal control group,PF+si-NC group and PF+si-PAI-1 group.Mice in PF+si-NC group and PF+si-PAI-1 group were intraperitoneally injected with 300μl si-NC or si-PAI-1(0.1 mg/kg).The positive expression of PAI-1,CollagenⅠandα-SMA in mouse peritoneal tissue was detected by immunohistochemical staining.Results Compared with end-stage renal disease group,the expression level of PAI-1 in the peritoneal tissue was significantly increased in peritoneal dialysis group(P<0.05).Compared with normal control group,the relative expression levels of PAI-1,CollagenⅠ,α-SMA,p-Smad3,p-EGFR and p-STAT3 proteins in the peritoneal mesothelial cells were significantly increased in TGF-β1+si-NC group(P<0.05),while E-cadherin and Acetylα-tubulin levels were significantly reduced(P<0.05).Compared with TGF-β1+si-NC group,the relative protein expression levels of PAI-1,CollagenⅠ,α-SMA,p-Smad3,p-EGFR and p-STAT3 in peritoneal mesothelial cells were decreased significantly in

关 键 词：腹膜透析 纤溶酶原激活物抑制因子1 腹膜纤维化 TGF-β1/Smad3 EGFR/STAT3 

分 类 号：R692[医药卫生—泌尿科学]