昼夜节律时钟基因3抑制棕榈酸诱导的足细胞氧化应激及炎症因子分泌  

Period circadian clock 3 inhibits palmitic acid-induced oxidative stress and inflammatory factor secretion in podocytes

作　　者：彭琳 张可可 陈科[2] PENG Lin;ZHANG Keke;CHEN Ke(Department of Nephrology,First Hospital of Changsha,Changsha 410005;Department of Endorcrinology,Third Xiangya Hospital,Central South University,Changsha 410013,China)

机构地区：[1]长沙市第一医院肾内科,长沙410005 [2]中南大学湘雅三医院内分泌科,长沙410013

出　　处：《中南大学学报（医学版）》2021年第11期1177-1186,共10页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(81870589)。

摘　　要：目的:高脂诱导足细胞损伤是导致肥胖相关性肾病(obesity relatedglomerulopathy,ORG)的重要因素之一,但机制并不明确。本研究探讨昼夜节律时钟基因3(period circadianclock 3,PER3)在抑制棕榈酸(palmitic acid,PA)诱导的小鼠足细胞氧化应激及炎症因子分泌中的作用和机制。方法:正常及高脂饮食喂养C57BL/6J小鼠16周,取小鼠肾组织,采用蛋白印迹法检测正常体重组和肥胖组PER3的表达;以不同体积浓度(0、50、150和300μmol/L)PA分别干预足细胞48 h,检测各浓度组PER3 mRNA及蛋白质的表达;以150μmol/L PA分别干预足细胞0、24、36及48 h,检测各时间点PER3 mRNA及蛋白质的表达;在足细胞中转染腺病毒(adenovirus,Ad)-PER3或小干扰RNA(small interferingRNA,siRNA)-PER3后48 h,再以150μmol/L PA干预48 h,检测PA组、Ad-PER3+PA组和siRNAPER3+PA组足细胞中三酰甘油(triglyceride,TG)的含量;利用RNA测序(RNA sequencing,RNA-seq)检测siRNA-PER3组和siRNA-对照组差异基因的表达;在足细胞中转染Ad-PER3或Ad-对照48 h后再以150μmol/L PA干预48 h,检测Ad-PER3+PA组和Ad-对照+PA组肾病蛋白(nephrin)、足蛋白(podocin)、足细胞标记蛋白(podocalyxin)、肾小球足突细胞膜黏蛋白(podoplanin)、超氧化物歧化酶1(superoxide dismutase 1,SOD1)、谷胱甘肽过氧化物酶1(glutathione peroxidase 1,GPX1)、过氧化氢酶(catalase,CAT)的表达,以及足细胞内丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)、肿瘤坏死因子-α(tumor necrosisfactor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)及白细胞介素-2(interleukin-2,IL-2)的含量。结果:在肥胖组小鼠肾组织中,PER3的表达较正常体重组小鼠下调(P<0.05);150μmol/L PA干预足细胞48 h较0、24、36 h PER3 mRNA表达均显著下调(均P<0.01);与PA组相比,Ad-PER3+PA组足细胞内TG含量显著减少,相反,siRNA-PER3+PA组足细胞内TG的含量增加(均P<0.05);在足细胞转染siRNA-PER3后,siRNObjective:High fat-induced podocyte injury is one of the important factors leading to obesity related nephropathy(ORG),but the mechanism is not clear.This study aims to explore the mechanism of period circadian clock 3(PER3)in the oxidative stress and inflammation induced by palmitic acid(PA)in podocytes.Methods:The C57BL/6J mice were fed with chow and high-fat diet for 16 weeks.The PER3 expression in kidney tissues were detected in the normal body weight group and the obesity group.The PER3 mRNA and protein expression were detected after the podocytes were induced with different concentrations(0,50,150 and 300μmol/L)of PA for 48 h.The PER3 mRNA and protein expression were detected after the podocytes were induced with 150μmol/L PA for 0,24,36,and 48 h.Triglyceride(TG)levels were examined in the PA group,the adenovirus(ad)-PER3+PA group,and the siRNA-PER+PA group after the podocytes were transfected by Ad-PER3 or small interfering RNA(siRNA)-PER3 for 48 h and subsequently were induced with 150μmol/L PA for 48 h.The differential gene expression was detected using RNA sequencing(RNA-seq)after podocytes were transfected by siRNA-PER3(siRNA-PER3 group)and siRNA-control(siRNA-control group),respectively.The mRNA levels of nephrin,podocin,podocalyxin,podoplanin,superoxide dismutase 1(SOD1),glutathione peroxidase 1(GPX1),catalase(CAT),and the levels of malondialdehyde(MDA),glutathione(GSH),tumor necrosis factorα(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β)and interleukin-2(IL-2)were detected after podocytes were transfected with Ad-PER3 or Ad-control for 48 h and then they were induced by 150μmol/L PA for 48 h.Results:The PER3 was down-regulated in the obesity group compared with the normal body weight group(P<0.05),and the PER3 was significantly down-regulated after the podocytes were treated with 150μmol/L for 48 h compared with 0,24,and 36 h(all P<0.01).The TG contents were significantly decreased in the Ad-PER3+PA group compared with the PA group(P<0.05).On the contrary,TG contents were increased in the

关 键 词：PER3 棕榈酸 足细胞 氧化应激 炎症因子 

分 类 号：R589.2[医药卫生—内分泌] R692[医药卫生—内科学]