肝豆汤Ⅱ号调控TGF-β1/Smad通路抑制肝豆状核变性小鼠肝纤维化的机制研究  

Study on the Mechanism of Gandou DecoctionⅡin Regulating TGF-β1/Smad Pathway to Inhibit Hepatic Fibrosis in Wilson Disease Mice

作　　者：周丹 董健健[1,2] 程楠 韩咏竹[1,2] ZHOU Dan;DONG Jianjian;CHENG Nan;HAN Yongzhu(Anhui University of Chinese Medicine,Hefei 230012,China;Institute of Neurology,Anhui University of Chinese Medicine,Hefei 230061,China)

机构地区：[1]安徽中医药大学,安徽合肥230012 [2]安徽中医药大学神经病学研究所,安徽合肥230061

出　　处：《中国中医药信息杂志》2024年第5期61-67,共7页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：安徽省自然科学基金(2208085MH226);安徽省高等学校科学研究项目(2023AH050778);合肥综合性国家科学中心大健康研究院新安医学与中医药现代化研究所“揭榜挂帅”项目(2023CXMMTCM002)。

摘　　要：目的研究肝豆汤Ⅱ号调控TGF-β1/Smad通路对肝豆状核变性小鼠肝纤维化的影响及机制。方法以DL小鼠为对照组,将TX小鼠随机分为模型组和中药低、高剂量组,中药组分别予常规剂量和高剂量肝豆汤Ⅱ号灌胃,对照组和模型组予蒸馏水灌胃,连续30 d。ELISA检测小鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、Ⅳ型胶原(ColⅣ)、透明质酸(HA)、Ⅲ型前胶原(HPCⅢ)、层粘连蛋白(LN)含量,Masson染色观察肝组织纤维化程度,免疫荧光染色检测肝组织α-平滑肌肌动蛋白(α-SMA)表达,电感耦合等离子体质谱法检测肝组织铜、铁离子含量,RT-qPCR检测肝组织α-SMA、Ⅰ型胶原(ColⅠ)、转化生长因子-β1(TGF-β1)及Smad2/3 mRNA表达,Western blot检测肝组织α-SMA、ColⅠ、TGF-β1、p-Smad2/3、Smad2/3蛋白表达。结果与对照组比较,模型组小鼠血清ALT、AST、ColⅣ、HA、HPCⅢ、LN含量均显著升高(P<0.05,P<0.01),肝组织胶原容积分数显著增加、α-SMA表达显著升高(P<0.01),肝组织铁离子含量显著升高(P<0.01),α-SMA、ColⅠ、TGF-β1、Smad2/3 mRNA和蛋白表达显著升高,p-Smad2/3蛋白表达显著升高(P<0.05,P<0.01);与模型组比较,中药低、高剂量组小鼠血清ALT、AST、ColⅣ、HA、HPCⅢ、LN含量均显著降低(P<0.05,P<0.01),肝组织胶原容积分数显著减少、α-SMA表达显著降低(P<0.01),肝组织铜和铁离子含量显著升高(P<0.01),α-SMA、ColⅠ、TGF-β1、Smad2/3mRNA和蛋白表达显著降低,p-Smad2/3蛋白表达显著降低(P<0.05,P<0.01)。结论肝豆汤Ⅱ号可通过调控TGF-β1/Smad通路下调ColⅠ、α-SMA和TGF-β1表达,抑制肝豆状核变性小鼠肝纤维化,减轻铜蓄积诱导的肝脏病理损伤。Objective To study the effects and mechanism of Gandou DecoctionⅡin regulating TGF-β1/Smad pathway for hepatic fibrosis in Wilson disease mice.Methods DL mice were set as the control group,while TX mice were randomly divided into the model group,TCM low-and high-dosage groups.TCM groups were given routine dosage and high-dosage Gandou DecoctionⅡby gavage respectively,while the control group and model group were administered distilled water by gavage for 30 consecutive days.ELISA was used to detect the contents of alanine aminotransferase(ALT),aspartate aminotransferase(AST),collagen typeⅣ(ColⅣ),hyaluronic acid(HA),procollagen typeⅢ(HPCⅢ),and laminin(LN)in serum,Masson staining was used to observe the degree of liver tissue fibrosis,while immunofluorescence staining was used to detectα-SMA expression in liver tissue,inductively coupled plasma mass spectrometry was used to detect copper and iron ion content in liver tissue,and RT-qPCR was used to detect the expressions ofα-SMA,collagen typeⅠ(ColⅠ),transforming growth factor-β1(TGF-β1),and Smad2/3 mRNA in liver tissue,Western blot was used to detect of protein expressions ofα-SMA,ColⅠ,TGF-β1,p-Smad2/3 and Smad2/3 in liver tissue.Results Compared with the control group,the contents of serum ALT,AST,ColⅣ,HA,HPCⅢand LN significantly increased in the model group(P<0.05,P<0.01),the collagen volume fraction of liver tissue significantly increased,the expression ofα-SMA significantly increased(P<0.01),the iron ion content in liver tissue significantly increased(P<0.01),α-SMA,ColⅠ,TGF-β1,Smad2/3 mRNA and protein expressions significantly increased,and the expression of p-Smad2/3 protein significantly increased(P<0.05,P<0.01).Compared with the model group,the contents of serum ALT,AST,ColⅣ,HA,HPCⅢand LN significantly decreased in TCM low-and high-dosage groups(P<0.05,P<0.01),the collagen volume fraction of liver tissue significantly decreased,the expression ofα-SMA significantly decreased(P<0.01),and the copper and iron ion content in l

关 键 词：肝豆状核变性 肝豆汤Ⅱ号 肝纤维化 TGF-β1/Smad通路 小鼠 

分 类 号：R285.5[医药卫生—中药学]