SIRT1调控Nrf2/HO-1信号通路在脓毒症肝损伤中的作用及其机制研究  

Role and mechanism of SIRT1 in regulating Nrf2/HO-1 signaling pathway in septic liver injury

作　　者：陈梦晓 张怡人 王毅[1,2,3] 古丽菲热·塔依尔[1,2,3] 于湘友 Chen Mengxiao;Zhang Yiren;Wang Yi;Gulifeire·Tayier;Yu Xiangyou(Critical Care Medicine Center,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang Uygur Autonomous Region,China;Xinjiang Medical University,Urumqi 830054,Xinjiang Uygur Autonomous Region,China;Xinjiang Key Laboratory of Medical Animal Model Research,Urumqi 830054,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]新疆医科大学第一附属医院重症医学中心,乌鲁木齐830054 [2]新疆医科大学,乌鲁木齐830054 [3]新疆医学动物模型研究重点实验室,乌鲁木齐830054

出　　处：《中华危重病急救医学》2023年第6期598-603,共6页Chinese Critical Care Medicine

基　　金：国家自然科学基金(82160360);新疆维吾尔自治区科技支疆项目(2021E02064)。

摘　　要：目的探讨沉默信息调节因子1(SIRT1)调控核因子E2相关因子2(Nrf2)/血红素加氧酶-1(HO-1)信号通路在脓毒症致肝损伤氧化应激和炎症反应中的作用与机制。方法采用随机数字表法将24只雄性SD大鼠分为假手术(Sham)组、盲肠结扎穿孔术(CLP)组、SIRT1激动剂SRT1720预处理(CLP+SRT1720)组、SIRT1抑制剂EX527预处理(CLP+EX527)组,每组6只。术前2 h分别向CLP+SRT1720组和CLP+EX527组腹腔注射10 mg/kg的SRT1720或EX527。于制模后24 h腹主动脉取血并处死大鼠取肝脏组织,用酶联免疫吸附试验(ELISA)检测血清中白细胞介素(IL-6、IL-1β)和肿瘤坏死因子-α(TNF-α)水平;用微板法检测血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平;用苏木素-伊红(HE)染色观察各组大鼠病理损伤情况;使用相应试剂盒分别检测肝组织丙二醛(MDA)、8-羟基脱氧鸟苷(8-OHdG)、谷胱甘肽(GSH)含量及超氧化物歧化酶(SOD)活性;采用实时荧光定量聚合酶链反应(RT-qPCR)和蛋白质免疫印迹试验(Western blotting)检测肝组织SIRT1、Nrf2、HO-1的mRNA及蛋白表达。结果与Sham组比较,CLP组血清IL-6、IL-1β、TNF-α、ALT、AST水平明显升高;组织病理学结果显示肝索排列紊乱,肝细胞肿胀坏死,出现大量炎症细胞浸润;肝组织中MDA、8-OHdG含量升高,GSH含量及SOD活性降低,SIRT1、Nrf2、HO-1的mRNA及蛋白表达水平明显下降,提示脓毒症大鼠出现肝功能障碍,肝组织内SIRT1、Nrf2、HO-1以及抗氧化蛋白水平降低,而氧化应激和炎症水平升高。与CLP组比较,CLP+SRT1720组大鼠炎症因子和氧化应激水平明显降低,SIRT1、Nrf2、HO-1的mRNA及蛋白表达水平显著升高〔IL-6(ng/L):34.59±4.21比61.84±3.78,IL-1β(ng/L):41.37±2.70比72.06±3.14,TNF-α(ng/L):76.43±5.23比130.85±5.30,ALT(U/L):30.71±3.63比64.23±4.59,AST(U/L):94.57±6.08比145.15±6.86,MDA(μmol/g):6.11±0.28比9.23±0.29,8-OHdG(ng/L):117.43±10.38比242.37±11.71,GSH(μmol/g):11.93±0.88比7.66±0.47,SODObjective To investigate the role and mechanism of silent information regulator 1(SIRT1)in regulating nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway in oxidative stress and inflammatory response to sepsis-induced liver injury.Methods A total of 24 male Sprague-Dawley(SD)rats were randomly divided into sham operation(Sham)group,cecal ligation and puncture(CLP)group,SIRT1 agonist SRT1720 pretreatment(CLP+SRT1720)group and SIRT1 inhibitor EX527 pretreatment(CLP+EX527)group,with 6 rats in each group.Two hours before operation,SRT1720(10 mg/kg)or EX527(10 mg/kg)were intraperitoneally injected into the CLP+SRT1720 group and CLP+EX527 group,respectively.Blood was collected from the abdominal aorta at 24 hours after modeling and the rats were sacrificed for liver tissue.The serum levels of interleukins(IL-6,IL-1β)and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).The serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected by microplate method.Hematoxylin-eosin(HE)staining was used to observe the pathological injury of rats in each group.The levels of malondialdehyde(MDA),8-hydroxydeoxyguanosine(8-OHdG),glutathione(GSH)and superoxide dismutase(SOD)in liver tissue were detected by corresponding kits.The mRNA and protein expressions of SIRT1,Nrf2 and HO-1 in liver tissues were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blotting.Results Compared with the Sham group,the serum levels of IL-6,IL-1β,TNF-α,ALT and AST in the CLP group were significantly increased;histopathological results showed that liver cords were disordered,hepatocytes were swollen and necrotic,and a large number of inflammatory cells infiltrated;the contents of MDA and 8-OHdG in liver tissue increased,while the contents of GSH and SOD decreased;and the mRNA and protein expressions of SIRT1,Nrf2 and HO-1 in liver tissues were significantly decreased.These results suggest that sepsis rats have liver dysfunc

关 键 词：脓毒症 肝损伤 沉默信息调节因子1 核因子E2相关因子2/血红素加氧酶-1信号通路 氧化应激 

分 类 号：R459.7[医药卫生—急诊医学] R575[医药卫生—治疗学]