咖啡酸苯乙酯通过TGF-β1/Smad3通路抗肝纤维化作用的研究

doi:10.3760/cma.j.cn441417-20250317-15007

国际医药卫生导报 ›› 2025, Vol. 31 ›› Issue (15): 2497-2502.DOI: 10.3760/cma.j.cn441417-20250317-15007

咖啡酸苯乙酯通过TGF-β1/Smad3通路抗肝纤维化作用的研究

杨宁¹ 王瑞平² 张亚密¹ 李亚萍³ 党双锁³

¹陕西中医药大学附属医院肿瘤医院二病区，咸阳 712000；²陕西中医药大学附属医院肝病医院二病区，咸阳 712000；³西安交通大学第二附属医院感染科，西安 710004

收稿日期:2025-03-17 出版日期:2025-08-01 发布日期:2025-08-21
通讯作者: 王瑞平，Email：543995687@qq.com
基金资助:
陕西省自然科学基金（2022SF-316）；秦创原产业聚集区联合基金（172）；陕西中医药大学附属医院科研课题（2020MS010）

Research on the anti-hepatic fibrosis effect of caffeic acid phenethyl ester via the TGF-β1/Smad3 pathway

Yang Ning¹, Wang Ruiping², Zhang Yami¹, Li Yaping³, Dang Shuangsuo³

¹Department of Cancer Ward 2,Shaanxi University of Traditional Chinese Medicine Affiliated Hospital, Xianyang 712000, China; ²Department of Infectious Disease Ward 2, Shaanxi University of Traditional Chinese Medicine Affiliated Hospital, Xianyang 712000, China; ³Department of Infectious Disease, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, China

Received:2025-03-17 Online:2025-08-01 Published:2025-08-21
Contact: Wang Ruiping,Email:543995687@qq.com
Supported by:
Natural Science Foundation of Shaanxi Province (2022SF-316); Qinchuang Original Industry Cluster Joint Fund (172); Scientific research project of Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine (2020MS010)

摘要/Abstract

摘要：

目的　通过动物实验研究咖啡酸苯乙酯（CAPE）对肝星状细胞（HSC）的活化、细胞外基质沉积等方面的作用，进而从TGF-β1/Smad3通路较为深入地探讨CAPE抗肝纤维化作用及分子机制。方法　实验时间为2021年月9至2022年9月，在陕西中医药大学创新平台进行。根据前期动物实验结果，选用雄性SD（SD）大鼠35只分为空白对照组（10只）、四氯化碳（CCl₄）造模组（15只）、CAPE干预组（10只），体质量（254.50±16.22）g。干预10周后行后续实验；通过电镜观察大鼠肝脏组织；通过HE、Masson、弹力纤维、网状纤维染色观察肝组织病理变化。采用实时聚合酶链反应（real-time PCR）、免疫组织化学和蛋白质印迹（Western blot）检测肝组织中平滑肌肌动蛋白α（α-SMA）、Ⅰ型胶原蛋白（Collagen Ⅰ）、转化生长因子β1（TGF-β1）、p-Smad3、组织金属蛋白酶抑制物1（TIMP1）、基质金属蛋白酶2（MMP2）mRNA水平和蛋白水平的表达情况。统计学方法采用单因素方差分析及Dunnett-t检验。结果　电镜下CAPE干预组可见细胞结构完整，未观察到簇状增生细胞肌丝的分布。HE、Masson、弹力纤维、网状纤维染色结果显示，CAPE干预组的肝脏组织胶原水平较CCl₄造模组减轻。免疫组织化学染色结果显示：CCl₄造模组与空白对照组相比，肝脏组织中的Collagen Ⅰ、α-SMA、TGF-β1和p-Smad3表达水平均显著增加（均P<0.05）；而CAPE干预组上述指标的表达水平均明显降低（均P<0.05）。western-blot和real-time PCR检测结果与之一致。但MMP2的表达水平在CCl₄造模组中明显降低，在CAPE干预组中明显增加（均P<0.05）。结论　CAPE可明显改善CCl₄复合因素所致的肝脏病理损害，逆转肝纤维化进程。CAPE可能通过抑制大鼠HSC活化以及胶原的形成，调节TIMP1/MMP2平衡，下调TGF-β1/Smad3信号通路的表达，从而发挥抗肝纤维化的作用。

关键词:

肝纤维化, 咖啡酸苯乙酯, 肝星状细胞

Abstract:

Objective　To investigate the effects of caffeic acid phenethyl ester (CAPE) on the activation of hepatic stellate cells (HSCs), extracellular matrix deposition, and to explore the anti-hepatic fibrosis effects and molecular mechanisms of CAPE through the TGF-β1/Smad3 pathway. Methods　The experiment was conducted on the innovation platform of Shaanxi University of Traditional Chinese Medicine from September 2021 to September 2022. Based on preliminary animal experiment results, 35 male Sprague-Dawley (SD) rats were selected and divided into a blank control group (10 rats), a carbon tetrachloride (CCl₄) model group(15 rats), and a CAPE intervention group(10 rats), with an average body weight of (254.50±16.22) g. After 10 weeks of intervention, subsequent experiments were conducted. Electron microscopy was used to observe liver tissue, and HE, Masson, elastic fiber, and reticular fiber staining were performed to assess pathological changes in liver tissue. Real-time polymerase chain reaction (real-time PCR), immunohistochemistry, and Western blotting were used to detect the expression levels of α-smooth muscle actin (α-SMA), type I collagen (Collagen I), transforming growth factor β1 (TGF-β1), p-Smad3, tissue inhibitor of metalloproteinases 1 (TIMP1), and matrix metalloproteinase 2 (MMP2) mRNA and protein levels in liver tissue. Statistical analysis was performed using one-way ANOVA and Dunnett's t test. Results　Electron microscopy revealed that in the CAPE intervention group, cell structures were intact, and no clustered proliferative cell myofibrils were observed. Staining results from HE, Masson, elastic fibers, and reticular fibers showed that collagen levels in the liver tissue of the CAPE intervention group were reduced compared to the CCl₄ model group. Immunohistochemical staining results indicated that the expression levels of Collagen I, α-SMA, TGF-β1, and p-Smad3 in liver tissue were significantly increased in the CCl₄ model group compared to the blank control group (all P<0.05), while CAPE intervention significantly reduced their expression levels in liver tissue (all P<0.05). Western-blot and real-time PCR results were consistent. However, MMP2 showed opposite results in mRNA and protein expression. Conclusion　CAPE can significantly improve liver pathological damage caused by CCl₄combined factors and reverse the progression of liver fibrosis. CAPE may exert its anti-fibrotic effects by inhibiting the activation of HSCs and collagen formation, regulating the TIMP1/MMP2 balance, and downregulating the expression of the TGF-β1/Smad3 signaling pathway.

Key words:

Liver fibrosis, Caffeic acid phenethyl ester, hepatic stellate cell

杨宁王瑞平张亚密李亚萍党双锁.

咖啡酸苯乙酯通过TGF-β1/Smad3通路抗肝纤维化作用的研究 [J]. 国际医药卫生导报, 2025, 31(15): 2497-2502.

Yang Ning, Wang Ruiping, Zhang Yami, Li Yaping, Dang Shuangsuo.

Research on the anti-hepatic fibrosis effect of caffeic acid phenethyl ester via the TGF-β1/Smad3 pathway [J]. International Medicine and Health Guidance News, 2025, 31(15): 2497-2502.

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咖啡酸苯乙酯通过TGF-β1/Smad3通路抗肝纤维化作用的研究

Research on the anti-hepatic fibrosis effect of caffeic acid phenethyl ester via the TGF-β1/Smad3 pathway

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