干酪乳杆菌培养上清液对小鼠急性肝损伤的保护作用及机制研究  

Protective effect of lactobacillus casei supernatant on CCl4-induced acute liver injury in mice and the underlying mechanisms

作　　者：周颖 张谢[2] 许丰 马燕燕 吴俊男 邵初晓[4] ZHOU Ying;ZHANG Xie;XU Feng;MA Yanyan;WU Junnan;SHAO Chuxiao(Department of Gastroenterology,Ningbo Medical Center LiHuili Hospital,Ningbo 315040,China;不详)

机构地区：[1]宁波市医疗中心李惠利医院消化内科,315040 [2]宁波市医疗中心李惠利医院药剂科,315040 [3]温州医科大学精神医学学院 [4]丽水市中心医院肝胰胆管外科

出　　处：《浙江医学》2022年第5期459-465,共7页Zhejiang Medical Journal

基　　金：宁波市自然基金项目(2021J286、202003N4235);浙江省自然科学基金资助(LY19H030004)。

摘　　要：目的探讨干酪乳杆菌培养上清液(LC-cs)对四氯化碳(CCl4)诱导的小鼠急性肝损伤的保护作用及相关机制。方法将24只ICR小鼠分为CCl4组、LC-cs+CCl4组(给予含LC-cs的饮用水2周)和对照组,每组8只。2周后前两组小鼠腹腔注射0.2%CCl4以诱导小鼠急性肝损伤;24 h后处死取肝组织;计算小鼠肝脏指数,测定血浆ALT、AST和高迁移率族蛋白B1(HMGB1)水平,评估肝脏组织病理学改变;测定肝脏组织炎症因子IL-6和TNF-α蛋白和mRNA水平;检测肝脏组织凋亡细胞,并测定凋亡相关蛋白的B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(BAX)表达水平;测定氧化应激指标包括谷胱甘肽过氧化物酶(GSH-Px)、总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平和NF-E2相关因子(Nrf2)蛋白表达水平;测定内质网应激相关蛋白包括葡萄糖调节蛋白78(GRP78)、转录活化因子6(ATF6)和C/EBP同源蛋白(CHOP)表达水平。结果CCl4组小鼠肝脏指数、血浆ALT、AST和HMGB1水平均高于对照组,LC-cs+CCl4组小鼠上述指标均低于CCl4组,差异均有统计学意义(均P<0.05)。CCl4组的肝脏组织病理切片出现了脂肪变性、炎细胞浸润、小叶中心充血及坏死,LC-cs+CCl4组的肝损伤改变轻于CCl4组。CCl4组IL-6和TNF-α的蛋白及mRNA水平高于对照组,LC-cs+CCl4组IL-6和TNF-α蛋白及mRNA水平低于CCl4组,差异均有统计学意义(均P<0.05)。CCl4组存在大量亮绿荧光的凋亡细胞,LC-cs+CCl4组的凋亡细胞明显少于CCl4组。CCl4组BAX表达高于对照组,Bcl-2表达低于对照组,LC-cs+CCl4组BAX表达低于CCl4组,Bcl-2表达高于CCl4组,差异均有统计学意义(均P<0.05)。CCl4组GSH-Px、T-AOC和SOD水平低于对照组,MDA水平高于对照组,LC-cs+CCl4组的GSH-Px、T-AOC和SOD水平高于CCl4组,MDA水平低于CCl4组,差异均有统计学意义(均P<0.01)。CCl4组GRP78、ATF6和CHOP蛋白表达水平高于对照组,而LC-cs+CCl4组GRP78、ATF6和CHOP蛋白表达水平低于CCl4组,差�Objective To explore the effect of lactobacillus casei supernatant(LC-cs)on carbon tetrachloride(CCl4)-induced acute liver injury in mice and the underlying mechanisms.Methods Twenty-four ICR mice were randomized into CCl4 group,LC-cs+CCl4 group and control group.Acute liver injury was induced by intraperitoneal injection of 0.2% CCl4 in CCl4 and LC-cs+CCl4 groups,mice in LC-cs+CCl4 group were given drinking water with LC-cs for 2 weeks before CCl4 injection.Mice were sacrificed after 24h of modelling.Liver weight/body weight ratio was calculated.Plasma alanine aminotransferase(ALT),aspartate transaminase(AST)and High Mobility Group Box 1(HMGB1)was measured.Histopathological features of mice liver tissue were examined.The expression levels of IL-6 and TNF-α protein and mRNA in liver tissue were measured.Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL)assay was used to count apoptotic cell numbers.Bcl-2 associated X protein(BAX)and B cell lymphoma 2(Bcl-2)protein levels were measured.Oxidative stress markers including glutathione peroxidase(GSH-Px),total anti-oxidant capacity(T-AOC),superoxide dismutase(SOD),malondialdehyde(MDA)and nuclear factor erythroid-2 related factor 2(Nrf2)protein level were measured.Endoplasmic reticulum stress related proteins,including glucose regulated protein 78(GRP78),activating transcription factor 6(ATF6)and C/EBP Homologous Protein(CHOP)were measured.Results Compared with control group,liver weight/body weight ratio,plasma ALT,AST and HMGB1 levels,hepatocyte necrosis and inflammation,IL-6 and TNF-α levels increased significantly in CCl4 group(P<0.05),however,compared to CCl4 group these indicators in LC-cs+CCl4 group were significantly decreased(P<0.01).CCl4 group had increased apoptotic cell number and BAX expression,and decreased Bcl-2 expression compared to control group(P<0.05),while compared to CCl4 group,the apoptotic cell number and BAX expression were decreased and Bcl-2 expression was increased in LC-cs+CCl4 group(P<0.05).CCl4 group had signif

关 键 词：干酪乳杆菌 四氯化碳 急性肝损伤 

分 类 号：R575[医药卫生—消化系统]