摘要
目的探讨白藜芦醇苷介导沉默信息调节因子1(SIRT1)抑制未折叠蛋白反应-凋亡途径在肾缺血再灌注损伤中的作用。方法32只大鼠随机分为4组:假手术组(A组)、肾缺血再灌注组(B组)、白藜芦醇苷组(C组)和SIRT1抑制剂EX-527组(D组)。后3组应用夹闭双侧肾动脉的方法构建肾缺血再灌注损伤模型,C及D组在造模前分别给予白藜芦醇苷10 mg/(kg·d)及EX-5275 mg/(kg·d)经尾静脉注射,持续1周。采用血清生化检查检测肌酐(Cr)、血尿素氮(BUN);采用TUNEL法检测细胞凋亡,硫代巴比妥酸法及WST-1法分别检测细胞内超氧化物歧化酶(SOD)以及丙二醛(MDA)含量,2',7'-二氯二氢荧光素二乙酸酯(DCFH-DA)负载法检测活性氧(ROS)的含量;采用ELISA法检测白细胞介素(IL)-1β和肿瘤坏死因子-α(TNF-α)含量;Western blot法检测SIRT1、X盒结合蛋白1(XBP1s)、转录激活子4(ATF4)及半胱天冬蛋白酶-3(caspase-3)的蛋白水平。结果与A组比较,B组血清Cr、BUN、MDA、ROS含量和细胞凋亡指数升高,SOD含量降低,IL-1β、TNF-α、SIRT1、XBP1s、ATF4及caspase-3表达上调;与B组比较,C组Cr、BUN、MDA、ROS含量和细胞凋亡指数降低,同时IL-1β、TNF-α、SIRT1、XBP1s、ATF4及caspase-3表达也下降,而SOD含量升高;D组通过下调SIRT1后各项指标变化与C组呈相反的趋势(P<0.05)。结论白藜芦醇苷可通过上调SIRT1表达清除氧化应激产物及减轻炎性反应来保护肾缺血再灌注损伤,其可能机制与其抑制未折叠蛋白反应-凋亡信号通路有关。
Objective To investigate the effects of resveratrol glycoside-mediated SIRT1 inhibition of unfolded protein response-apoptosis pathway on renal ischemia-reperfusion injury.Methods Thirty-two rats were randomly divided into four groups,sham operation group(Group A),renal ischemia-reperfusion group(Group B),resveratrol glycoside group(Group C)and SIRT1 inhibitor EX-527 group(Group D).The model of renal ischemia-reperfusion injury was established by bilateral renal arteries clamping in the latter three groups.The Group C and D recieved injection of resveratrol glycoside 10 mg/(kg·d)and EX-5275 mg/(kg·d),respectively,through caudal vein for one week before modeling.Serum biochemical examination was used to detect creatinine(SCr)and blood urea nitrogen(BUN).TUNEL method was used to detect apoptosis.The contents of superoxide dismutase(SOD)and malondialdehyde(MDA)were assessed by thiobarbituric acid method and WST-1 method,respectively.The content of reactive oxygen species(ROS)was assessed by DCFH-DA loading method.The contents of IL-1βand TNF-a were assessed by ELISA method.The protein levels of SIRT1,XBP1s,ATF4 and caspase-3 were assessed by Western blot.Results Compared with Group A,the contents of Cr,BUN,MDA,ROS and apoptosis index in Group B were significantly increased,while the content of SOD was significantly reduced and the expression of IL-1β,TNF-a,SIRT1,XBP1s,ATF4 and caspase-3 was significantly up-regulated(P<0.05).Compared with Group B,the contents of Cr,BUN,MDA,ROS and apoptosis index were significantly reduced in Group C,while the expression of IL-1β,TNF-a,SIRT1,XBP1s,ATF4 and caspase-3 was significantly down-regulated,but the content of SOD was significantly increased(P<0.05).The protective effect of resveratrol glycoside on renal ischemia-reperfusion injury was significantly inhibited in Group D.Conclusion Resveratrol glycoside can protect renal ischemia-reperfusion injury by up-regulating SIRT1 expression to clear oxidative stress products and attenuating inflammatory response,which may be correlated to its inhibition of unfolded protein response-apoptosis signal pathway.
作者
谢志娟
陈建英
XIE Zhi-juan;CHEN Jian-ying(Department of Nephrology,the First Affiliated Hospital of Nanhua University,Hengyang 421001,Hunan,China)
出处
《广东医学》
CAS
2020年第12期1215-1218,共4页
Guangdong Medical Journal
基金
国家自然科学基金资助项目(81903615)
湖南省自然科学基金资助项目(2018JJ2014)。
关键词
白藜芦醇苷
凋亡
未折叠蛋白反应
肾缺血再灌注损伤
resveratrol glycoside
apoptosis
unfolded protein response
renal ischemia-reperfusion injury
