尿毒症晚期糖基化终产物和蛋白结合溶质通过抑制Krüppel-样因子2诱导腹膜间皮细胞功能障碍的机制研究  

The mechanism of advanced glycation end products and protein-binding solutes on peritoneal mesothelial dysfunction by inhibition of Krüppel-like factor 2 in uremia.

作　　者：黄德胜 庄宇亮 HUANG De-sheng;ZHUANG Yu-liang(The eighth ward,Nanan hospital,fujian provinee,Fujian 362300,China;The ninth ward,Nanan hospital,fujian province,Fujian 362300,China)

机构地区：[1]福建省南安市医院,362300

出　　处：《首都食品与医药》2020年第5期14-16,共3页Capital Food Medicine

摘　　要：目的探讨尿毒症晚期糖基化终产物(AGEs)和蛋白结合溶质(AOPP)通过抑制Krüppel-样因子2(KLF2)诱导腹膜间皮细胞(PMC)功能障碍的机制。方法采用MTT法分别检测不同浓度的AGEs和AOPP对人腹膜间皮细胞的抑制率;采用实时定量PCR(q-PCR)法测定AGEs和AOPP对人腹膜间皮细胞中KLF2和转化生长因子β1(TGF-β1)的mRNA表达;采用蛋白免疫印迹(Western Blot)法测定AGEs和AOPP对人腹膜间皮细胞中上皮-间叶转化(EMT)相关蛋白Ⅰ型胶原(Collagen I)、α平滑肌肌动蛋白(α-SMA)和E-钙黏蛋白(E-cadherin)的蛋白表达。结果不同浓度的AGEs(0~120 mM)和AOPP(0~120 mM)均对人腹膜间皮细胞的增殖具有一定的抑制作用,并且80mM AGEs和60 mMAOPP的抑制效果最显著(P<0.01),后续实验将人腹膜间皮细胞分组为正常对照(Control)组、80mM AGEs处理(AGEs)组和60mM AOPP处理(AOPP)组。与正常对照组相比,AGEs组和AOPP组人腹膜间皮细胞中KLF2的mRNA表达降低(P<0.01)。与正常对照组相比,AGEs组和AOPP组人腹膜间皮细胞中TGF-β1的mRNA表达增加(P<0.01)。与正常对照组相比,AGEs组和AOPP组人腹膜间皮细胞中Collagen I、α-SMA的蛋白表达上调(P<0.01),而E-cadherin的蛋白表达下调(P<0.01)。结论尿毒症晚期糖基化终产物和蛋白结合溶质均可通过抑制Krüppel-样因子2上调TGF-β1、Collagen I和α-SMA的表达,下调E-cadherin的表达,诱导人腹膜间皮细胞的上皮-间叶转化,进而导致人腹膜间皮细胞功能障碍。Objective In this study,the mechanism of advanced glycation end products (AGEs) and protein-binding solutes (AOPP) on peritoneal mesothelial dysfunction by inhibition of Krüppel-like factor 2 (KLF2) in uremia was investigated.Methods The MTT assay was used to detect the inhibition rate of AGEs and AOPP at different concentrations on HPMC.A real-time quantitative PCR (q-PCR) method was used to determine the mRNA expressions of KLF2 and transformed growth factor β1 (TGF-β1) in human peritoneal mesothelial cells by AGEs or AOPP treatment.Western Blot method was used to determine the protein expressions of EMT related proteins Collagen type Ⅰ (Collagen I),α smooth muscle actin (α-SMA) and E-cadherin in human peritoneal mesothelial cells by AGEs or AOPP treatment.Results Different concentrations of which (0~120mM) and AOPP (0~120mM) on peritoneal mesothelium cell proliferation has certain inhibition,which and 60mM and 80mM AOPP inhibitory effect is the most significant (P<0.01),follow-up experiments to peritoneal mesothelium cell group as normal Control (Control) group,the 80mM (which) which treatment group and 60mM AOPP treatment group (AOPP).Compared with the control group,the mRNA expression of KLF2 was decreased in the AGEs group and AOPP group (P<0.01).Compared with the control group,mRNA expression of TGF-beta 1 in peritoneal mesothelial cells in the AGEs group and AOPP group increased (P<0.01).Compared with the normal control group,Collagen I and alpha-sma protein expressions in the AGEs group and AOPP group were up-regulated (P<0.01),while the protein expression of e-cadherin was down-regulated (P<0.01).Conclusion Uremia advanced glycosylation end products and protein-binding solutes all could up-regulate the expressions of TGF-β1,Collagen I and α-SMA,down-regulate the expression of E-cadherin,induce the EMT of human peritoneal mesothelial cells,eventually leading to dysfunction of peritoneal mesothelial cells.

关 键 词：晚期糖基化终产物 蛋白结合溶质 Krüppel-样因子2 上皮-间叶转化 

分 类 号：R692.5[医药卫生—泌尿科学]