SRT1720对对乙酰氨基酚诱导小鼠急性肝损伤的保护作用及其机制  

Protective effect of SRT1720 on acute liver injury induced by acetaminophen in mice and its mechanism

作　　者：邹巧玲 袁秋林 宋银宏[1,2,3] ZOU Qiaoling;YUAN Qiulin;SONG Yinhong(Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy,China Three Gorges University,Yichang 443002,Hubei Province,China;不详)

机构地区：[1]三峡大学肿瘤微环境与免疫治疗湖北省重点实验室,湖北宜昌443002 [2]三峡大学感染与炎症损伤研究所,湖北宜昌443002 [3]三峡大学医学院,湖北宜昌443002

出　　处：《中国生物制品学杂志》2024年第2期188-194,共7页Chinese Journal of Biologicals

基　　金：国家自然科学基金面上项目(81671397);湖北省卫健委重点项目(WJ2019H528)。

摘　　要：目的 评价沉默信息调节因子2-相关酶1(silent information regulator 2-related enzymes1,SIRT1)激活剂(SRT1720)对对乙酰氨基酚(acetaminophen,APAP)诱导小鼠肝损伤的保护作用,并探讨其作用机制。方法 将雄性C57BL/6J小鼠随机分为对照、SRT1720、APAP和APAP+SRT1720组,每组10只。SRT1720和APAP+SRT1720组小鼠经灌胃给予SRT1720(30 mg/kg体质量),对照和APAP组经灌胃给予等体积生理盐水,均每天给药1次,连续给药5 d;第6天,APAP和APAP+SRT1720组小鼠分别经腹腔注射APAP(325 mg/kg体质量),对照和SRT1720组小鼠经腹腔注射等体积生理盐水。24 h后,摘除眼球取外周全血,分离血清,采用相应试剂盒检测丙氨酸氨基转移酶(alanine aminotransferase,ALT)及门冬氨酸氨基转移酶(aspartate aminotransferase,AST)水平;将小鼠经断颈处死,无菌取肝脏组织,HE染色法观察肝组织病理改变,RT-qPCR法检测PERK-eIF2α-CHOP信号通路相关分子GRP78、PERK、eIF2α、ATF4、CHOP基因mRNA转录水平,Western blot法检测上述基因相应蛋白及Caspase12蛋白的相对表达水平。结果与对照组比较,APAP组小鼠血清中ALT和AST水平均明显上升(t分别为55.21和34.29,P均<0.01);肝组织中可见明显大片状肝细胞坏死,肝小叶结构发生显著改变,部分区域肝细胞发生肿胀变形较严重;GRP78、PERK、eIF2α、ATF4和CHOP基因的mRNA转录及蛋白相对表达水平均明显增高(t=9.85~33.89,P均<0.05);Caspase12蛋白相对表达水平显著升高(t=11.78,P <0.01)。与APAP组比较,APAP+SRT1720组小鼠血清中ALT和AST水平均明显降低(t分别为42.92和18.02,P均<0.01);肝细胞损伤程度明显减轻,发生肿胀变形的细胞也显著减少;GRP78、PERK、eIF2α、ATF4、CHOP基因的mRNA转录和蛋白相对表达水平均明显下降(t=6.19~22.43,P均<0.05);Caspase12蛋白表达水平未发生明显下降(t=0.34,P> 0.05)。结论 SRT1720可改善APAP诱导小鼠发生的肝损伤,可能是通过抑制内质网应激(endoplasmic reticulum strObjective To evaluate the protective effect of the activator of silent information regulator 2-related enzymes 1(SIRT1),SRT1720,on liver injury induced by acetaminophen(APAP)in mice and explore its mechanism.Methods Forty male C57BL/6J mice were randomly divided into normal control group,SRT1720 treatment group,APAP treatment group,and APAP+SRT1720 treatment group,with 10 mice in each group.Mice in SRT1720 and APAP+SRT1720 groups were given SRT1720(30 mg/kg body mass)by intragastric administration,while normal saline of equal volume was given by intragastric administration in control and APAP groups,once a day for 5 days;On the 6th day,mice in APAP and APAP+SRT1720 groups were injected i.p.with APAP(325 mg/kg body mass),while those in control and SRT1720 groups with equal volume of normal saline.After 24 hours,the peripheral blood was taken and the serum was separated,which were detected for the levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)by the corresponding kits;The liver tissue of mice was taken aseptically,observed for the pathological changes by HE staining,detected for the mRNA transcription levels of GRP78,PERK,eIF2α,ATF4 andCHOP genesrelatedtoPERK-eIF2α-CHOPsignalingpathwaybyRT-qPCR anddetected for the relative expression levels of these corresponding proteins and Caspase12 protein by Western blot.Results Compared with normal control group,the serum ALT and AST levels of mice in APAP group significantly increased(t=55.21 and 34.29 respectively,each P<0.01);significant necrosis of hepatocytes was observed in liver tissue,the structure of hepatic lobules changed significantly,and the swelling and deformation of hepatocytes in some areas were serious;the mRNA transcription and relative protein expression levels of GRP78,PERK,eIFα,ATF4 and CHOP genes increased significantly(t=9.85~33.89,each P<0.05)and the relative expression level of Caspase12 protein increased significantly(t=11.78,P<0.01).Compared with APAP group,the serum ALT and AST levels of mice in APAP+SRT1720 group de

关 键 词：沉默信息调节因子2-相关酶1激活剂 SRT1720 对乙酰氨基酚 急性肝损伤 PERK-eIF2α-CHOP信号通路 

分 类 号：R363.2[医药卫生—病理学]