丹参酚酸改善心肌纤维化的潜在机制研究  

Mechanism research of salvianolic acids treating myocardial fibrosis

作　　者：马耀磊 周凤洁 李霄 霍小位[4] 雷伟 MA Yaolei;ZHOU Fengjie;LI Xiao;HUO Xiaowei;LEI Wei(Institute of Traditional Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;State Key Laboratory of Component-based Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;State Key Laboratory of Chinese Medicine Modernization,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;College of Pharmaceutical Sciences,Hebei University,Baoding 071002,China)

机构地区：[1]天津中医药大学中医药研究院,天津301617 [2]天津中医药大学组分中药国家重点实验室,天津301617 [3]天津中医药大学现代中药创制全国重点实验室,天津301617 [4]河北大学药学院,保定071002

出　　处：《天津中医药》2025年第5期621-629,共9页Tianjin Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82104434);中国博士后科学基金(2022T150477,2021M702465)。

摘　　要：[目的]心肌纤维化可破坏心脏结构,导致心功能不全,影响心力衰竭患者的预后。现有药物对心肌纤维化的改善效果有限,而丹参作为活血化瘀药在心血管疾病防治中应用广泛。本研究旨在通过体外细胞实验和网络药理学分析,探讨丹参酚酸对心肌纤维化的抑制作用及其机制。[方法]采用体外实验结合网络药理学方法。体外实验中,在10 ng/mL转化生长因子-β1(TGF-β1)诱导大鼠心肌成纤维细胞表型转化模型中和TGF-β1/血管紧张素Ⅱ(AngⅡ)诱导的H9c2细胞模型中,分别以25~100μg/mL丹参酚酸干预,通过实时聚合酶链式反应(q-PCR)和蛋白免疫印记法(Western blot)探讨丹参酚酸抑制心肌纤维化的药理作用;网络药理学分析中,对于质谱鉴定结果和心肌纤维化靶点进行整合分析。[结果]在TGF-β1刺激下,丹参酚酸可剂量依赖性地减少Ⅲ型胶原蛋白(COLIII)、I型胶原蛋白(COLI)和α-平滑肌肌动蛋白(α-SMA)表达,抑制心肌成纤维细胞表型转化。此外,丹参酚酸还可抑制TGF-β1/AngⅡ诱导的H9c2细胞中Ⅰ型胶原α-1链(COLIA1)、Ⅲ型胶原蛋白α-1链(COL3A1)、基质金属蛋白酶2(MMP2)和基质金属蛋白酶9(MMP9)的高表达。超高效液相色谱(UPLC)-四极杆飞行时间质谱(Q-TOF-MS)鉴定出丹参酚酸的24个化学成分,网络药理学分析显示其与心肌纤维化相关的靶基因主要富集在脂质和动脉硬化过程、磷脂酰肌醇3激酶(PI3K)-蛋白激酶B(AKT)信号通路等。[结论]通过分子生物学实验、UPLC-Q-TOF-MS和网络药理学分析,明确了丹参酚酸抑制心肌成纤维细胞表型转化和减少H9c2细胞促纤维化因子分泌的作用,揭示了其药效物质和可能机制,为丹参酚酸治疗心肌纤维化提供了研究依据。[Objective]Myocardial fibrosis can disrupt cardiac structure,leading to heart failure and adversely affecting the prognosis of patients with heart failure.Existing treatments have limited efficacy in improving myocardial fibrosis.Danshen,a traditional herb known for its blood-activating and stasis-eliminating properties,is widely used in the prevention and treatment of cardiovascular diseases.This study aims to investigate the inhibitory effects of salvianolic acid(a major active compound from Danshen)on myocardial fibrosis and its underlying mechanisms through in vitro cell experiments and network pharmacology analysis.[Methods]This study employed a combination of in vitro experiments and network pharmacology.In the in vitro experiments,salvianolic acid(25 to 100μg/mL)was applied to rat cardiac fibroblasts undergoing phenotypic transformation induced by 10 ng/mL TGF-β1 and to H9c2 cells stimulated by TGF-β1/AngⅡ.The pharmacological effects of salvianolic acid on myocardial fibrosis were explored using q-PCR and Western blot.In the network pharmacology analysis,the mass spectrometry results and myocardial fibrosis targets were integrated for comprehensive analysis.[Results]Under TGF-β1 stimulation,salvianolic acid dose-dependently reduced the expression of COLIII,COLI,andα-SMA,inhibiting the phenotypic transformation of cardiac fibroblasts.Additionally,salvianolic acid suppressed the overexpression of COLIA1,COL3A1,MMP2,and MMP9 in H9c2 cells induced by TGF-β1/AngⅡ.UPLC-Q-TOF-MS identified 24 chemical components in salvianolic acid.Network pharmacology analysis revealed that the target genes related to myocardial fibrosis were mainly enriched in lipid and atherosclerosis processes,as well as the PI3K-AKT signaling pathway.[Conclusion]This study,through molecular biology experiments,UPLC-Q-TOF-MS,and network pharmacology analysis,clarified the effects of salvianolic acid in inhibiting the phenotypic transformation of cardiac fibroblasts and reducing the secretion of pro-fibrotic factors in H9c2 cells.It

关 键 词：心肌纤维化 丹参酚酸 胶原沉积 活性机制 

分 类 号：R542.23[医药卫生—心血管疾病]