湖南中医药大学学报2025，Vol.45Issue(11)：2072-2080,9.DOI:10.3969/j.issn.1674-070X.2025.11.007
鳖龙软肝汤通过下调TGF-β1/Smad信号通路关键分子表达抑制肝癌的作用机制
Mechanism of Bielong Ruangan Decoction in inhibiting hepatocellular carcinoma via downregulation of key molecular expressions in the TGF-β1/Smad signaling pathway
摘要
Abstract
Objective To investigate the pharmacological mechanism by which Bielong Ruangan Decoction(BLRGD)inhibits hepatocellular carcinoma through modulation of the transforming growth factor-β1(TGF-β1)/Smad signaling pathway.Methods Sixty male SPF Sprague-Dawley(SD)rats were randomly divided into four groups:normal group,model group,low-dose BLRGD group(6.84 g·kg-1·d-1),and high-dose BLRGD group(27.36 g·kg-1·d-1),with 15 rats per group.Except the rats in the normal group,the remaining rats received intraperitoneal injection of diethylnitrosamine(DEN)(50 mg·kg-1·w-1)to establish a hepatocellular carcinoma model,with continuous intervention for 16 weeks.In Week 8,the low-and high-dose BLRGD groups received the corresponding doses(6.84,27.36 g·kg-1·d-1)of BLRGD for 8 consecutive weeks.Gener al condition was observed in each group.Pathological changes in liver tissue were assessed using hematoxylin-eosin(HE)staining.The protein expression levels of TGF-β1,Smad2,Smad3,lysyl oxidase-like protein 1(LOXL1),type I collagen(CollagenⅠ),tissue inhibitor of metalloproteinase 1(TIMP1),and alpha-smooth muscle actin(α-SMA)were measured by Western blot and immunofluorescence assays.The expression level of proliferating cell nuclear antigen(PCNA)was determined by immunohistochemistry.Results Rats in the normal group exhibited good general condition,with normal mental status,food consumption and activity.At Week 16,rats in the model group were lethargic,had dull fur,and showed a significant reduction in food consumption and activity.No differences were observed between the low-dose BLRGD group and the model group,while the high-dose BLRGD group showed improved mental status and increased food consumption.Hepatocytes in the normal group were normal,and the hepatic lobular structure was intact;atypical hepatocellular carcinoma cells were observed in the model group at Week 16;hepatocyte necrosis and inflammation were alleviated in the low-dose BLRGD group,while in the high-dose BLRGD group,hepatocyte arrangement was more regular with markedly reduced necrosis and degeneration.Compared with the normal group,the model group at Week 16 exhibited decreased body weight(P＜0.05),increased liver-to-body weight ratio(P＜0.05),and increased Ishak scores(P＜0.05);the expression levels of PCNA in liver tissue increased(P＜0.05),along with the increased protein and mRNA expression levels of TGF-β1,Smad2,Smad3,LOXL1,CollagenⅠ,TIMP1,and α-SMA(P＜0.05).Compared with the model group and the low-dose BLRGD group,the high-dose BLRGD group at Week 16 showed a decreased liver-to-body weight ratio(P＜0.05),and reduced PCNA expression levels(P＜0.05).Compared with the model group,the low-dose BLRGD group showed decreased protein expression levels of TGF-β1,TIMP1,and α-SMA in liver tissue(P＜0.05),and reduced mRNA expression levels of TGF-β1,Smad3,LOXL1,and TIMP1(P＜0.05);the high-dose BLRGD group showed decreased protein and mRNA expression levels of TGF-β1,Smad2,Smad3,LOXL1,CollagenⅠ,TIMP1,and α-SMA in liver tissue(P＜0.05).Compared with the low-dose BLRGD group,the high-dose BLRGD group showed further decreased expression levels of Smad2 and α-SMA proteins in liver tissue(P＜0.05),as well as reduced mRNA expression levels of Smad2,Smad3,and CollagenⅠ(P＜0.05).Conclusion BLRGD can ameliorate hepatic fibrosis and inhibit the progression of hepatocellular carcinoma in rats,and its mechanism may be related to the downregulation of PCNA and key molecules in the TGF-β1/Smad signaling pathway in liver tissue.关键词
肝细胞癌/肝纤维化/鳖龙软肝汤/转化生长因子-β1/信号转导蛋白通路/增殖细胞核抗原/二乙基亚硝胺/细胞外基质Key words
hepatocellular carcinoma/hepatic fibrosis/Bielong Ruangan Decoction/TGF-β1/Smad signaling pathway/proliferating cell nuclear antigen/diethylnitrosamine/extracellular matrix分类
中医学引用本文复制引用
周意,李倩,彭憬,孙克伟,银思涵..鳖龙软肝汤通过下调TGF-β1/Smad信号通路关键分子表达抑制肝癌的作用机制[J].湖南中医药大学学报,2025,45(11):2072-2080,9.基金项目
湖南省自然科学基金项目(2024JJ9442) （2024JJ9442）
湖南省中医药管理局科研计划项目(D2022059) （D2022059）
国家中医药管理局高水平中医药重点学科建设项目(zyyzdxk-2023146) （zyyzdxk-2023146）
湖南中医药大学双一流学科建设项目(2018[03]) （2018[03]）
湖南省卫生健康科研课题(20257775). （20257775）
