摘要
目的探讨2-十二烷基-6-甲氧基-2,5-二烯-1,4-环己二酮(2-dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione,DMDD)对CCl_(4)致大鼠肝纤维化的影响及潜在作用机制。方法采用50%CCl_(4)复制肝纤维化模型,检测血液中门冬氨酸转移酶(aspartate transferase,AST)、丙氨酸转移酶(alanine transferase,ALT)、白蛋白/球蛋白(albumin/globulin,A/G)、总蛋白(total protein,TP)、总胆红素(total bilirubin,T-BIL)、大鼠透明质酸(hyaluronic acid,HA)、层黏连蛋白(laminin,LN)、Ⅲ型胶原(collagen typeⅢ,ColⅢ)和Ⅳ型胶原(collagen typeⅣ,ColⅣ)。HE和Masson染色观察肝脏组织病变和纤维形成情况。免疫组织化学法检测肝脏组织中α平滑肌肌动蛋白(αsmooth muscle actin,α-SMA)、Ⅰ型胶原(collagen typeⅠ,ColI)、转化生长因子β1(transformed growth factor-β1,TGF-β1)、Smad2、Smad7蛋白表达水平。RT-PCR法检测肝脏组织中α-SMA、ColI、TGF-β1、Smad7mRNA水平。Western blot法检测肝脏组织中TGF-β1、Smad4、Smad7蛋白表达水平。结果与正常对照组相比,模型组血清AST、ALT、TP、T-BIL、HA、LN、ColⅢ、ColⅣ水平明显升高,A/G水平明显降低,肝脏HE和Masson染色显示有大量纤维形成,说明造模成功。与模型组相比,DMDD给药组大鼠AST、ALT、TP、T-BIL、HA、LN、ColⅢ、ColⅣ含量和α-SMA、ColI、TGF-β1、Smad2、Smad4mRNA及蛋白水平显著下调,A/G、Smad7水平上调。肝脏HE和Masson染色结果显示纤维增生明显减少。结论DMDD对CCl_(4)诱导的肝纤维化具有保护作用,其机制可能与TGF-β1/Smads信号通路有关。
Aim To investigate the effects of 2-dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione(DMDD)on resisting hepatic fibrosis induced by carbon tetrachloride(CCl_(4))in rats and the underlying mechanisms,with a specific focus on the TGF-β1/Smads signaling pathway.Methods The hepatic fibrosis model was replicated using 50%CCl_(4).Various parameters,including levels of aspartate transferase(AST),alanine transferase(ALT),albumin/globulin(A/G),total protein(TP),total bilirubin(T-BIL),hyaluronic acid(HA),laminin(LN),collagen typeⅢ(ColⅢ),and collagen typeⅣ(ColⅣ)in the blood,were measured.Liver tissue lesions and fiber formation were observed using HE and Masson staining.The expression levels ofαsmooth muscle actin(α-SMA),collagen typeⅠ(ColⅠ),transformed growth factor(TGF-β1),Smad2,and Smad7 proteins were assessed using immunohistochemistry.α-SMA,ColI,TGF-β1,and Smad7 mRNA levels in liver tissue were measured by RT-PCR.Additionally,the expression levels of TGF-β1,Smad4,and Smad7 proteins in liver tissue were determined by Western blot.Results In comparison to the normal control group,the model group exhibited significantly elevated levels of AST,ALT,TP,T-BIL,HA,LN,ColⅢand ColⅣin serum.But A/G level notably decreased.Successful modeling was confirmed by the presence of extensivefiber formations observed through HE and Masson staining in liver tissue.The DMDD administration group demonstrated a notable decrease levels of AST,ALT,TP,T-BIL,HA,LN,Col III,and CollV,but A/G was significantly elevated when compared to the model group.Furthermore,α-SMA,ColI,TGF-β1,Smad2 and Smad4 mRNA and protein levels in the DMDD administration group were significantly reduced,while Smad7 significantly declined.HE and Masson staining results reflected a marked reduction in fibrous hyperplasia.Conclusion DMDD exhibits a protective effect against CCl_(4)-induced hepatic fibrosis,and its mechanism appears to be associated with the TGF-β1/Smads signaling pathway.
作者
黄湘
方坤凤
范氏泰和
梁杏梅
黄仁彬
李学政
HUANG Xiang;FANG Kun-feng;Hoa Thi Thai Pham;LIANG Xing-mei;HUANG Ren-bin;LI Xue-zheng(Guangxi Health Science College,Nanning 530023,China;Pingnan Maternal and Child Health Hospital,Pingnan Guangxi 537300,China;Guangxi International Zhuang Medicine Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530001,China;School of Pharmacy,Guangxi Medical University,Nanning 530021,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2024年第3期545-551,共7页
Chinese Pharmacological Bulletin
基金
广西高校中青年教师(科研)基础能力提升项目(No 2020KY43009)
国家自然科学基金资助项目(No 81760665)。
