Drp1抑制剂Mdivi-1对脂多糖/D-半乳糖胺诱导的急性肝损伤的影响及机制研究  

Effects of Drp1 Inhibitor Mdivi-1 on Lipopolysaccharide/D-galactosamine-Induced Acute Liver Injury and Its Mechanism

作　　者：肖丽丹 唐俐 杨永强 李龙江 黄佳祎 沈宜 张力 XIAO Lidan;TANG Li;YANG Yongqiang;LI Longjiang;HUANG Jiayi;SHEN Yi;ZHANG Li(Department of Pathophysiology,College of Basic Medicine,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学,基础医学院病理生理学教研室,重庆400016

出　　处：《中国细胞生物学学报》2020年第3期392-399,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81871606)资助的课题。

摘　　要：线粒体动力相关蛋白1(dynamin-related protein 1,Drp1)是细胞凋亡调控的新靶点,而凋亡是急性肝损伤的重要特征,该研究采用选择性Drp1抑制剂Mdivi-1,探讨了Drp1在急性肝损伤中的可能病理生理学作用及其潜在的药靶价值。该研究动物实验采用6~8周龄雄性BAlB/c小鼠,在腹腔内注射脂多糖(lipopolysaccharide,LPS)/D-半乳糖胺(D-galactosamine,D-Gal),诱导急性肝损伤模型。实验分为四组:正常对照组(A)、Mdivi-1单独处理组(B)、模型组(C)和Mdivi-1干预组(D)。Mdivi-1在LPS/D-Gal暴露前30 min经腹腔注入,在LPS/D-Gal注射后1.5 h或6.0 h处死动物,采集肝组织和血浆标本。实验通过组织病理学染色、比色法检测血浆转氨酶活性、细胞因子TNF-α检测、casepase-3、casepase-8和casepase-9的活性检测和TUNEL技术探讨Mdivi-1介导下肝损伤的改善情况。结果显示,通过H&E染色观察肝组织病理学改变,正常对照组和Mdivi-1单独处理组小鼠肝组织结构无异常,LPS/D-Gal暴露后可引起一系列显著的组织学异常改变,Mdivi-1干预组肝组织学异常明显减轻。通过检测血浆谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)活性评估肝脏功能损害程度,Mdivi-1干预组ALT和AST的活性明显降低(P<0.05)。采用ELISA法检测血浆中肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)水平以评估炎症反应程度,Mdivi-1干预组的炎性细胞因子表达降低明显(P<0.05)。通过半胱氨酸蛋白酶-3(casepase-3)、半胱氨酸蛋白酶-8(casepase-8)、半胱氨酸蛋白酶-9(casepase-9)和TUNEL染色评估组织凋亡程度,Mdivi-1干预组的凋亡水平降低显著(P<0.05)。所以得出结论,Mdivi-1干预可减轻 LPS/D-Gal诱导的肝组织病变、降低血浆转氨酶、下调血浆中 TNF-α水平、降低肝内casepase-3、casepase-8和casepase-9活性并减少TUNEL阳性细胞的数量。以上数据表明,Drp1抑制剂Mdivi-1可减轻LPS/D-Gal诱导的急性�The mitochondrial protein Drp1 (dynamin-related protein 1) is a new target for the regulation of apoptosis,and apoptosis is an important feature of acute liver injury.In this study,the selective Drp1 inhibitor Mdivi-1 was used to explore the role of Drp1 in the possible pathophysiology of liver injury and its potential drug target value.In this study,6-8 week-old male BAlB/c mice were injected intraperitoneally with LPS (lipopolysaccharide)/D-Gal (D-galactosamine) to induce an acute liver injury model.The experiment was divided into four groups:normal control group (A),Mdivi-1 alone treatment group (B),model group (C),and Mdivi-1 intervention group (D).Mdivi-1 was injected intraperitoneally 30 minutes before LPS/D-Gal exposure.Animals were sacrificed after LPS/D-Gal injection in 1.5 h or 6.0 h,liver tissue and plasma samples were collected.Histopathological staining,colorimetric detection of plasma transaminase activity,detection of cytokine TNF-α,activity of casepase-3,casepase-8,and casepase-9 and TUNEL technology were used to explore the improvement of liver injury induced by Mdivi-1.The results showed that the pathological changes of liver tissue were observed by HE staining.There was no abnormal liver tissue structure in normal control group and Mdivi-1 alone treatment group.LPS/D-Gal exposure could cause a series of significant histological abnormalities.The degree of liver damage was assessed by measuring the activity of plasma ALT (alanine aminotransferase) and AST (aspartate aminotransferase).The activity of ALT and AST in the Mdivi-1 intervention group was significantly reduced (P<0.05).TNF-α (tumor necrosis factor alpha) levels in plasma were used to assess the degree of inflammatory response.The expression of inflammatory cytokines in the Mdivi-1 intervention group was significantly reduced (P<0.05).Casepase-3,casepase-8,casepase-9 and TUNEL staining were used to assess the degree of tissue apoptosis,and the level of apoptosis in the Mdivi-1 intervention group decreased significant (P<0.05).Therefor

关 键 词：动力相关蛋白1 线粒体 急性肝损伤 脂多糖 凋亡 

分 类 号：R575[医药卫生—消化系统]