摘要
目的探讨蛋白激酶C-α(PKC-α)基因在腹膜透析大鼠超滤衰竭中的作用。方法细胞水平检测下调PKC-α基因后血管内皮生长因子(VEGF)基因的表达量。将雄性SD大鼠分为正常对照组、假手术组、尿毒症组、腹膜透析2周组(PD2组)、腹膜透析4周组(PD4组)、腹膜透析4周后腹腔注射空质粒组(NC组)、腹膜透析4周后腹腔注射PKC-αsiRNA组(PKC-αsiRNA组)。处死前行腹膜平衡试验(PET),测定超滤量和葡萄糖转运量(MTG);Masson染色观察腹膜形态学变化;CD34免疫染色计数腹膜组织微血管密度(MVD),评估腹膜血管新生程度;检测各组大鼠腹膜组织PKC-αmRNA和蛋白表达水平。结果在细胞中下调PKC-α基因后,VEGF表达量明显降低。与正常对照组相比,尿毒症、PD2、PD4、NC组和PKC-αsiRNA组大鼠腹膜功能明显降低,MVD明显增大,PKC-αmRNA和蛋白表达水平明显升高,差异有统计学意义(P<0.05)。与尿毒症组相比,PD2、PD4组和NC组大鼠腹膜组织MVD明显增大,PKC-αmRNA和蛋白表达水平明显升高,且PD4组和NC组变化更明显。与PD4组和NC组比,PKC-αsiRNA组大鼠腹膜功能明显好转,MVD明显减小,PKC-αmRNA和蛋白表达水平明显降低。结论有可能通过下调VEGF基因的表达量,延缓超滤衰竭进程;通过改善透析液生物相容性、药物治疗或基因转染,以期减少细胞因子对治疗的影响。
Objective To investigate the role of protein kinase C-α(PKC-α)gene during the ultrafiltration failure process in peritoneal dialysis rats.Methods The vascular endothelial growth factor(VEGF)gene expression level was detected in the cellular level.Male SD rats were divided into the normal group,sham operation group and uremic group.And the peritoneal dialysis groups included the non-treatment group,peritoneal dialysis for 2 weeks group(PD2 group),peritoneal dialysis for 4 weeks group(PD4 group),PD4 injected with empty plasmid group(NC group)and PKC-αsiRNA group(PKC-α siRNA group).The peritoneal balance test(PET)was performed before sacrifice,the ultrafiltration amount and glucose transport amount(MTG)were determined.Then the peritoneal morphological changes were observed by the Masson staining.The peritoneal microvessel density(MVD)was counted by the CD34 immunostaining,and the degree of peritoneal angiogenesis was evaluated.The expression levels of PKC-αmRNA and protein in rat peritoneal tissues were detected in each group.Results The VEGF expression level was significantly decreased after down-regulating intracellular PKC-αgene.Compared with the normal control group,the peritoneal function in the uremic group,PD2,PD4,NC and PKC-αsiRNA groups was decreased significantly,while MVD and the expression level of PKC-αmRNA and protein in peritoneal tissues were increased significantly,and the differences were statistically significant(P<0.05).Compared with the uremic group,the MVD and the expression level of PKC-αmRNA and protein in peritoneal tissues in the PD2,PD4 and NC groups were increased significantly,moreover,these changes were more significant in the PD4 and NC groups.Compared with the PD4 and NC groups,the peritoneal function in the PKC-αsiRNA group was improved significantly,MVD and the expression level of PKC-αmRNA and protein were significantly decreased.Conclusion The ultrafiltration failure process may be delayed through down-regulating the VEGF gene expression level;improving the dialysate bi
作者
李玲麟
毕丹青
杨艳萍
何杰
袁红伶
LI Linglin;BI Danqing;YANG Yanping;HE Jie;YUAN Hongling(First Affiliated Hospital of Kunming Medical University,Kunming,Yunnan 650023,China)
出处
《重庆医学》
CAS
2019年第21期3601-3607,共7页
Chongqing medicine
基金
国家自然科学基金地区基金项目(81460649,81360662)
云南省科技厅应用基础研究项目(2013FZ)
云南省内设机构项目(2014NS229)
云南省医学领军人才项目(L-201606)
关键词
蛋白激酶C-Α
血管内皮生长因子
超滤衰竭
腹膜透析
腹膜平衡试验
微血管密度
protein kinase C-alpha
vascular endothelial growth factors
ultrafiltration failure
peritoneal dialysis
peritoneal balance test
microvessel density
