腹膜透析流出液中腹膜间皮细胞热休克蛋白47的表达及意义

The Expression of Heat Shock Protein 47 in Human Peritoneal Mesothelial Cells from Patients with CAPD

目的:探讨腹膜透析流出液中培养的人腹膜间皮细胞(human peritoneal mesothelial cells,HPMCs)热休克蛋白47(heat shock protein,HSP47)的表达,并分析其与腹膜间皮细胞转分化(epithelial-to-mesenchymal transition,EMT)的关系。方法:取桂林医学院附属医院门诊随诊的连续非卧床腹膜透析(continuous ambulatory peritoneal dialysis,CAPD)患者,按腹透时间分为两组:(1)新置管组(PD≤30 d),(2)腹透1年以上组(PD≥1年),取其夜间留腹腹透液分离培养的腹膜间皮细胞,采用Western-blot方法检测HSP47、E-cadherin、α-SMA的蛋白表达,分析HSP47表达与E-cadherin、α-SMA表达的相关性。结果:随着腹膜透析时间的延长,腹膜间皮细胞由圆形或椭圆形的上皮细胞形态逐渐变成梭形的纤维样细胞形态,与新置管组比较,PD 1年以上组腹透液腹膜间皮细胞HSP47的表达和α-SMA均同步增强,而PD一年以上组E-cadherin蛋白明显降低。两组细胞HSP47表达与α-SMA表达呈正相关(P <0. 05),HSP47表达与E-cadherin表达呈负相关(P <0. 05)。结论:HSP47随着腹膜透析时间的延长表达上调,可能参与腹膜间皮细胞EMT而导致腹膜纤维化。

Objective: Determine the HSP47 expression in HPMCs from effluent derived from patients undergoing CAPD and analysis the relationship between HSP47 and EMT associated proteins. Methods: We randomly selected patients with PD from the affiliated hospital of Guilin Medical University. Patients will be divided into two groups: Group A: CAPD start; Group B: CAPD over a year. Human peritoneal mesothelial cells( HPMCs) were cultured from effluents in dialysis fluid from patients undergoing CAPD. After 14 days,the cultured cells were identified by invert microscope and Western blot of HSP47,E-cadherin and α-smooth muscle actin( α-SMA). Results: HPMCs from effluent derived of patients undergoing CAPD had markedly varied morphologic features,ranging from a cobblestone-like appearance similar to that of mesothelium derived from omentum to fibroblast-like cells or mixed cell populations. By western blot,HSP47 was up-regulated expression in HPMCs from effluent derived of patients undergoing CAPD over a year,compared with those undergoing CAPD start( P < 0. 05). The results of Western blot showed that there was a markedly lower level of E-cadherin expression in HPMCs from effluent derived of patients undergoing CAPD start than those undergoing CAPD over a year( P < 0. 05). The level of α-SMA in HPMCs from effluent derived of patients undergoing CAPD over a year was higher than those undergoing CAPD start by western blot( P < 0. 05). There was a negative correlation between the expressions of HSP47 and E-cadherin protein of HPMCs from effluent derived of patients undergoing CAPD( r =-0. 996,P < 0. 05). There was a positive correlation between the expressions of HSP47 and α-SMA protein of HPMCs from effluent derived of patients undergoing CAPD( r =0. 980,P < 0. 05). Conclusion: The expression of HSP47 was increased appearantly in HPMCs from effluent derived of patients undergoing CAPD with the prolongation of PD. HSP47 may participate in human peritoneal mesothelial cells EMT and accelerate the peritoneal fibrosis.