阿司匹林阻断高脂血症诱导的足细胞内质网应激的机制  

Aspirin intervenes in hyperlipidemia kidney damage by blocking endoplasmic reticulum stress in podocytes

作　　者：褚宇东[1] 李荣山[2] 田渊 徐鹏杰[1] 刘江[1] 裘晓蕙[1] 步世忠[3] Chu Yudong;Li Rongshan;Tian Yuan;Xu Pengjie;Liu Jiang;Qiu Xiaohui;Bu Shizhong(Department of Nephrology,Ningbo Medical Center Lihuili Hospital,Ningbo 315000,China;Department of Nephrology,Shanxi Provincial People's Hospital,Taiyuan 030000,China;Diabetes Research Center,Ningbo University,Ningbo 315000,China)

机构地区：[1]宁波市医疗中心李惠利医院肾内科,宁波315000 [2]山西省人民医院肾内科,太原030000 [3]宁波大学糖尿病研究中心,宁波315000

出　　处：《中华肾脏病杂志》2020年第2期139-144,共6页Chinese Journal of Nephrology

基　　金：国家自然科学基金面上项目(81370165);宁波市科技惠民项目(201701CX-D02072);宁波市自然科学基金(2015A610217)。

摘　　要：目的探讨阿司匹林(aspirin,ASP)对高脂血症诱导的足细胞内质网应激的影响及可能机制。方法培养足细胞分为4组:对照组、ASP(100μg/ml)组、ox-LDL(100μg/ml)组和ASP+ox-LDL组,在6 h、12 h、24 h、48 h以实时定量PCR法检测蛋白激酶R样内质网激酶(protein kinase R-1ike endoplasmic reticulum kinase,PERK)、真核细胞起始因子2α(eukaryotic translation initiation factor 2α,eIF2α)、活化转录因子4(activating transcription factor-4,ATF4)、CCAAT/增强子结合蛋白同源蛋白(CAAT/enhancer binding protein homologous protein,CHOP)的表达;在24 h时以Western印迹法检测磷酸化(p)-PERK、p-eIF2α的表达;在12 h时以Western印迹法检测ATF4的表达。结果在足细胞培养24 h时,ox-LDL组和ASP+ox-LDL组PERK、eIF2α的表达水平达高峰,在12 h时,该两组ATF4、CHOP的表达水平达高峰。在足细胞培养6 h、12 h、24 h、48 h时,与对照组比较,ox-LDL组PERK、eIF2α、ATF4、CHOP的表达水平明显较高(均P<0.05);与ox-LDL组比较,ASP+ox-LDL组上述各指标表达明显较低(均P<0.05)。在足细胞培养24 h时,与对照组比较,ox-LDL组p-PERK、p-eIF2α的表达水平明显较高(均P<0.05);与ox-LDL组比较,ASP+ox-LDL组p-PERK、p-eIF2α的表达水平明显较低(均P<0.05)。在足细胞分组培养12 h时,各组ATF4蛋白水平的表达与mRNA表达相似。ASP组与对照组间上述各项指标表达差异无统计学意义。结论高脂血症可能通过诱导PERK和eIF2α的磷酸化、激活ATF4转录及诱导CHOP高表达,引起足细胞内质网应激,而阿司匹林可能部分阻断了PERK通路,进而可能对足细胞具有保护作用。Objective To investigate the effects and underlying mechanisms of aspirin on endoplasmic reticulum stress in podocytes induced by hyperlipemia.Methods Cultured podocytes were divided into four groups:control group,aspirin(100μg/ml)group,oxidized low density lipoprotein(ox-LDL,100μg/ml)group,aspirin+ox-LDL group.The expression of protein kinase R-1ike endoplasmic reticulum kinase(PERK),eukaryotic translation initiation factor 2α(eIF2α),activating transcription factor-4(ATF4)and CAAT/enhancer binding protein homologous protein(CHOP)at 6 h,12 h,24 h,48 h were evaluated by real-time PCR.The related proteins of p-PERK and p-eIF2αat 24 h and ATF4 at 12 h were evaluated by Western blotting,respectively.Results The expressions of PERK,eIF2αpeaked at 24 h,while ATF4 and CHOP peaked at 12 h in ox-LDL group and aspirin+ox-LDL group.Compared with control group,the expressions of PERK,eIF2α,ATF4 and CHOP were significantly higher in ox-LDL group at each times(all P<0.05).Compared with ox-LDL group,the expressions of the above indicators were significantly lower in aspirin+ox-LDL group at each times(all P<0.05).At 24 h,compared with control group,the expressions of p-PERK and p-eIF2αwere significantly higher in ox-LDL group(both P<0.05).Compared with ox-LDL group,the expressions of p-PERK and p-eIF2αwere significantly lower in aspirin+ox-LDL group(both P<0.05).At 12 h,the expression of ATF4 protein in each group was similar to that of mRNA.There were no significant difference in the expressions of all indicators between aspirin group and control group.Conclusions Hyperlipidemia may cause endoplasmic reticulum stress in podocytes by inducing phosphorylation of PERK and eIF2α,activating ATF4 transcription and inducing high expression of CHOP.Aspirin may partially block the PERK pathway,which may have protective effects for podocytes.

关 键 词：阿司匹林 高脂血症 脂蛋白类 LDL 足细胞 内质网应激 

分 类 号：R589[医药卫生—内分泌]