南方医科大学学报

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (9): 1946-1958.doi: 10.12122/j.issn.1673-4254.2025.09.14

• • 上一篇    

肉桂酸通过抑制TLR4减轻阿霉素诱导的小鼠心肌损伤铁死亡的发生

云琦1,2(), 杜若丽3,4, 贺玉莹1,2, 张贻欣5, 王佳慧3,4, 叶红伟3,4, 李正红1,2, 高琴3,4()   

  1. 1.蚌埠医科大学，生命科学学院，安徽 蚌埠 233030
    2.蚌埠医科大学，安徽省神经再生技术与医用新材料工程研究中心，安徽 蚌埠 233030
    3.蚌埠医科大学，心脑血管疾病基础与临床重点实验室，安徽 蚌埠 233030
    4.蚌埠医科大学，生理学教研室，安徽 蚌埠 233030
    5.蚌埠医科大学，临床医学院，安徽 蚌埠 233030
  • 收稿日期:2025-02-13 出版日期:2025-09-20 发布日期:2025-09-28
  • 通讯作者: 高琴 E-mail:yunqiyq1009@163.com;bbmcgq@126.com
  • 作者简介:云 琦，硕士，E-mail: yunqiyq1009@163.com
  • 基金资助:
    国家级创新创业训练计划项目(202410367058);蚌埠医科大学研究生科研创新项目(Byycx23003);安徽省优秀科研创新团队项目(2022AH010083)

Cinnamic acid ameliorates doxorubicin-induced myocardial injury in mice by attenuating cardiomyocyte ferroptosis via inhibiting TLR4

Qi YUN1,2(), Ruoli DU3,4, Yuying HE1,2, Yixin ZHANG5, Jiahui WANG3,4, Hongwei YE3,4, Zhenghong LI1,2, Qin GAO3,4()   

  1. 1.School of Life Sciences, Bengbu Medical University, Bengbu 233030, China
    2.Anhui Engineering Research Center for Neural Regeneration Technology and Medical New Materials, Bengbu Medical University, Bengbu 233030, China
    3.Key Laboratory of Basic and Clinical Cardiovascular and Cerebrovascular Diseases, Bengbu Medical University, Bengbu 233030, China
    4.Department of Physiology, Bengbu Medical University, Bengbu 233030, China
    5.School of Clinical Medicine, Bengbu Medical University, Bengbu 233030, China
  • Received:2025-02-13 Online:2025-09-20 Published:2025-09-28
  • Contact: Qin GAO E-mail:yunqiyq1009@163.com;bbmcgq@126.com

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摘要：

目的 基于网络药理学技术结合动物实验探讨肉桂酸（CA）改善阿霉素心肌损伤（DIC）的可能机制。 方法 采用网络药理学分析得到CA改善DIC的关键靶点，并通过分子对接进行验证。6~8周龄SPF级雄性C57BL/6J小鼠被随机分为5组（6只/组）：Sham组、阿霉素（DOX）组、CA（25 mg/kg）+DOX组、CA（50 mg/kg）+DOX组、CA（100 mg/kg）+DOX和铁死亡抑制剂（Ferrostatin-1 5 mg/kg）+DOX组。超声心动图和HE染色观察心肌组织病理变化；检测小鼠血清 CK-MB、LDH、MDA、IL-6、TNF-α和心肌ROS水平。蛋白免疫印迹检测心肌组织中TLR4及铁死亡通路相关蛋白表达水平。构建慢病毒介导的基因稳定敲低TLR4的小鼠心肌HL-1细胞株，细胞分为6组：阴性对照组（NC）；DOX模型组（DOX）；铁死亡诱导剂组（Erastin）；低表达组（si-TLR4组）；DOX+si-TLR4组；Erastin+si-TLR4组。CCK8测定细胞活力；荧光探针检测细胞ROS含量变化；免疫荧光检测GPX4蛋白的表达。 结果 通过网络药理学分析获得交集靶点28个，KEGG富集分析显示，CA可能通过Toll样受体信号通路作用于DIC。动物实验显示，与Sham组相比，DOX组超声心动图和HE 染色结果显示心脏损伤，血清CK-MB、LDH、MDA、炎症因子和心肌ROS水平升高（P<0.01），SLC7A11和GPX4蛋白表达降低，TLR4和PTGS2蛋白表达升高（P<0.05）。经CA处理后，心超功能指标水平升高（P<0.05）；血清 CK-MB、LDH、MDA、炎症因子和心肌ROS水平降低（P<0.05）；CA和铁死亡抑制剂使心肌组织SLC7A11和GPX4蛋白表达升高，TLR4和PTGS2蛋白表达降低（P<0.05）。低表达TLR4细胞株中，与NC组相比，DOX组和Erastin组细胞，ROS水平升高（P<0.001），GPX4蛋白表达降低（P<0.001）；与DOX组和Erastin组相比，DOX+si-TLR4组和Erastin+si-TLR4组细胞ROS水平降低（P<0.05），GPX4蛋白表达升高（P<0.01）。 结论 CA减轻小鼠DIC，其机制可能与抑制TLR4减轻铁死亡有关。

关键词: 肉桂酸, 阿霉素心肌损伤, 铁死亡, TLR4, 网络药理学

Abstract:

Objective To explore the mechanism of cinnamic acid (CA) for improving doxorubicin-induced myocardial injury (DIC) in mice. Methods Network pharmacology analysis was used to obtain the key targets of CA and DIC. Male C57BL/6J mice were randomized into Sham, DOX, CA (25, 50 and 100 mg/kg)+DOX, and CA+Ferrostatin-1+DOX groups, and their myocardial function and pathology were examined by echocardiography and HE staining. Serum levels of CK-MB, LDH, MDA, IL-6, TNF‑α and myocardial ROS level were detected, and the expression levels of TLR4 and ferroptosis pathway proteins in myocardial tissue were detected by Western blotting. Cultured murine cardiomyocytes (HL-1 cells) with or without transfection with a small interfering RNA targeting TLR4 (si-TLR4) were treated with DOX or Erastin, and the cellular ROS content was measured by DCFH-DA staining; the expression level of GPX4 was detected using immunofluorescence staining. Results Network pharmacology analysis suggested that CA may improve DIC through TLR4 signaling. DOX treatment caused obvious myocardial injury in mice, which showed significantly increased serum levels of CK-MB, LDH, MDA, IL-6, TNF-α and myocardial ROS level with decreased myocardial levels of SLC7A11 and GPX4 proteins and increased levels of TLR4 and PTGS2 proteins. All these changes in the mouse models were significantly alleviated by treatment with CA, and the mice receiving CA or ferrostatin-1 treatment exhibited increased myocardial expressions of SLC7A11 and GPX4 proteins and lowered expressions of TLR4 and PTGS2 proteins. In cultured HL-1 cells, treatment with DOX and Erastin both obviously increased intracellular ROS level and decreased cellular GPX4 expression level, and these changes were strongly attenuated by TLR4 interference. Conclusion CA, as a potent herbal monomer, can effectively alleviate DIC in mice by inhibiting TLR4-mediated ferroptosis.

Key words: cinnamic acid, doxorubicin-induced myocardial injury, ferroptosis, Toll-like receptor 4, network pharmacology