五味子酯甲通过TLR4/NF-κB通路对肝纤维化的保护作用研究  

Protective Effect of Schisantherin A on Liver Fibrosis through TLR4/NF-κB Pathway

作　　者：王春梅[1] 车金营 崔凯[2] 徐博 庄文越 李贺[1] 王海莉[2] WANG Chunmei;CHE Jinying;CUI Kai;XU Bo;ZHUANG Wenyue;LI He;WANG Haili(Pharmacy College of Beihua University,Jilin 132013,China;Affiliated Hospital of Beihua University,Jilin 132011,China;Basic Medical College of Jilin Medical College,Jilin 132013,China;Medical Technology College of Beihua University,Jilin 132013,China)

机构地区：[1]北华大学药学院,吉林吉林132013 [2]北华大学附属医院,吉林吉林132011 [3]吉林医药学院基础医学院,吉林吉林132013 [4]北华大学医学技术学院,吉林吉林132013

出　　处：《北华大学学报（自然科学版）》2022年第6期757-762,共6页Journal of Beihua University（Natural Science）

基　　金：吉林省自然科学基金项目白求恩专项基金(20200201513JC).

摘　　要：目的观察五味子酯甲(Schisantherin A,SCA)对小鼠肝纤维化的保护作用,并初步探讨其作用机制.方法应用硫代乙酰胺(thioacetamide,TAA)诱导小鼠肝纤维化模型,SCA(4 mg/kg)灌胃给药治疗5周后检测小鼠血清中谷丙转氨酶(alanine transaminase,ALT)、谷草转氨酶(aspartate transaminase,AST)活性及肝组织中羟脯氨酸(hydroxyproline,Hyp)含量;ELISA法检测小鼠肝组织中肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)、白介素-6(interleukin-6,IL-6)、白介素-1β(interleukin-1β,IL-1β)含量;H&E及Masson染色检测小鼠肝组织损伤及胶原纤维表达情况;体外实验中,应用转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导人肝星状细胞(LX2)的活化,应用Western blot法检测细胞中α-平滑肌动蛋白(α-smooth muscle actin,α-SMA)、Toll样受体4(toll-like receptor 4,TLR4)、MyD88及核因子-κB(nuclear factor-κB,NF-κB)蛋白表达水平.结果与模型组比较,SCA给药组小鼠血清中ALT、AST活性、肝组织中Hyp、TNF-α、IL-6及IL-1β含量均显著降低(P<0.01),肝损伤明显减轻,肝组织胶原纤维的范围和面积明显减少.同时,SCA能明显抑制TGF-β1诱导的LX2细胞中α-SMA、TLR4、MyD88及NF-κB蛋白表达(P<0.05).结论SCA具有抗肝纤维化作用,其机制可能通过TLR4/NF-κB信号通路抑制肝星状细胞的活化及炎症反应发挥作用.Objective To observe the protective effect of Schisantherin A(SCA)on liver fibrosis in mice and explore its mechanism.Method A liver fibrosis model of mouse was constructed by the progressive intraperitoneal injection of thioacetamide(TAA),and SCA(4 mg/kg)was administered by gavage for 5 weeks.The activities of alanine transaminase(ALT),aspartate transaminase(AST)in serum and the content of hydroxyproline(Hyp)in liver tissue were measured;ELISA method was used to detect the content of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1βin liver tissue;Hematoxylin&eosin(H&E)and Masson stainings were employed to observe liver injury and collagen fiber expression in mice.In vitro,the activation of human hepatic stellate cells(LX2)was induced by transforming growth factor-β1(TGF-β1).The protein expressions ofα-s mooth muscle actin(α-SMA),toll-like receptor 4(TLR4),MyD88 and nuclear factor-κB(NF-κB)were detected by Western blotting.Results Compared with the model group,the activities of ALT and AST in serum,the contents of Hyp,TNF-α,IL-6 and IL-1βin liver tissue of mice were significantly decreased(P<0.01),the liver injury was significantly relived,and the range and area of collagen fibers in liver tissue were significantly reduced.Meanwhile,SCA significantly inhibited protein expressions ofα-SMA,TLR4,MyD88 and NF-κB in activated LX2 cells by TGF-β1(P<0.05).Conclusion SCA has anti-fibrosis effect,and its mechanism may inhibit hepatic stellate cell activation and inflammatory response through TLR4/NF-κB signaling pathway.

关 键 词：五味子酯甲 肝星状细胞 肝纤维化 TRL4 NF-ΚB 

分 类 号：R285[医药卫生—中药学]