中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (25): 5382-5389.doi: 10.12307/2025.529

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

芍药苷对血管紧张素Ⅱ诱导心肌成纤维细胞纤维化的保护作用

纪雅琼1，2，宁忠平1，2   

  1. 1上海中医药大学，上海市   201203；2上海健康医学院附属周浦医院心血管内科，上海市   201318
  • 收稿日期:2024-04-20 接受日期:2024-07-08 出版日期:2025-09-08 发布日期:2024-12-25
  • 通讯作者: 宁忠平，硕士，主任医师，上海中医药大学，上海市 201203；上海健康医学院附属周浦医院心血管内科，上海市 201318
  • 作者简介:纪雅琼，女，1997年生，山东省邹平市人，汉族，上海中医药大学在读硕士，主要从事中西医结合治疗心血管疾病方面的研究。
  • 基金资助:
    上海市浦东新区卫生健康委员会重点学科群建设项目(PWZxq2022-11)，项目负责人：宁忠平；上海市浦东新区心房颤动流行病学调查及分级诊疗模式下房颤全程管理前瞻性队列研究项目(PKJ2021-Y33)，项目负责人：宁忠平；上海市浦东新区卫健委临床高峰学科建设项目(PWYgf2021-04)，项目负责人：宁忠平

Protective effect of paeoniflorin on angiotensin II-induced fibrosis in cardiac fibroblasts

Ji Yaqiong1, 2, Ning Zhongping1, 2   

  1. 1Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; 2Department of Cardiology, Zhoupu Hospital Affiliated to Shanghai University of Medicine & Health Science, Shanghai 201318, China
  • Received:2024-04-20 Accepted:2024-07-08 Online:2025-09-08 Published:2024-12-25
  • Contact: Ning Zhongping, Master, Chief physician, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Department of Cardiology, Zhoupu Hospital Affiliated to Shanghai University of Medicine & Health Science, Shanghai 201318, China
  • About author:Ji Yaqiong, Master candidate, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Department of Cardiology, Zhoupu Hospital Affiliated to Shanghai University of Medicine & Health Science, Shanghai 201318, China
  • Supported by:
    Key Discipline Group Construction Project of Pudong New Area Health and Health Commission, No. PWZxq2022-11 (to NZP); Epidemiological Investigation of Atrial Fibrillation in Pudong New Area and Prospective Cohort Study on the Whole Process Management of Atrial Fibrillation under the Mode of Graded Diagnosis and Treatment, No. PKJ2021-Y33 (to NZP); Pudong New Area Health Committee Peak Discipline Construction, No. PWYgf2021-04 (to NZP)

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摘要：

文题释义：

芍药苷：是芍药总苷中的主要生物活性成分，具有抗氧化、抗炎、抗纤维化、降血脂、免疫调节和保肝的作用，大多被用于肝纤维化的治疗，此外，还可用于溃疡性结肠炎、类风湿性关节炎和心脏、肾脏、肝脏等器官缺血性疾病的治疗。
心肌纤维化：是一种以成纤维细胞异常增殖、胶原过度沉积及异常分布为特征的病理表现。当心肌细胞在缺血、缺氧、炎症以及毒素等损害作用下，整个细胞发生变性、坏死、凋亡，或者是心肌细胞在结构、生化以及代谢方面发生了变化，都会激活巨噬细胞等免疫细胞，生成和分泌多种活性生长因子，最终促使心肌成纤维细胞活化并大量增殖，分泌合成Ⅰ型和Ⅲ型胶原蛋白，来修复或替代受损的心肌细胞。

摘要
背景：研究表明芍药苷对肝、肾等器官纤维化具有改善作用，尤其是在肝纤维化中表现出突出优势，但芍药苷对于血管紧张素Ⅱ诱导的心肌纤维化的保护作用尚不明确。
目的：探讨芍药苷对血管紧张素Ⅱ诱导的心肌成纤维细胞的保护作用及分子机制。
方法：在分离培养的SD大鼠乳鼠心肌成纤维细胞中加入血管紧张素Ⅱ(1 μmol/L)干预48 h作为模型组；芍药苷低、高剂量组给予不同剂量的芍药苷(50，100 μmol/L)预处理2 h，再用血管紧张素Ⅱ处理48 h；SIRT1抑制剂组先用10 μmol/L SIRT1抑制剂EX527处理2 h，再用100 μmol/L芍药苷处理2 h，最后用血管紧张素Ⅱ处理48 h。采用CCK-8法检测细胞活力，Transwell检测细胞迁移能力，用DHA荧光探针检测细胞内活性氧水平，用试剂盒检测氧化应激标志物水平，Western blot检测纤维化相关基因的蛋白表达，qRT-PCR检测细胞外基质和纤维化相关基因的mRNA表达。
结果与结论：①与对照组相比，血管紧张素Ⅱ干预后心肌成纤维细胞的增殖、迁移能力明显提高，细胞内活性氧和丙二醛水平升高，超氧化物歧化酶和过氧化氢酶活性降低，α-平滑肌肌动蛋白、Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、纤维连接蛋白、结缔组织生长因子、基质金属蛋白酶9的mRNA表达增加；与模型组相比，芍药苷剂量依赖性抑制上述效应改变(P < 0.01)；②与模型组相比，芍药苷剂量依赖性上调SIRT1的蛋白表达(P < 0.001)；③与芍药苷高剂量组相比，SIRT1抑制剂组细胞迁移数量、α-平滑肌肌动蛋白、Ⅰ型胶原蛋白、Ⅲ型胶原蛋白表达水平显著增加(P < 0.01)。结果表明，芍药苷可能通过上调SIRT1的表达，有效减轻了血管紧张素Ⅱ诱导的心肌成纤维细胞纤维化改变，剂量依赖性地抑制了心肌成纤维细胞氧化应激和细胞外基质沉积，对于心肌成纤维细胞纤维化具有保护作用。

关键词: 芍药苷, 心肌成纤维细胞, 血管紧张素Ⅱ, 细胞外基质, 纤维化, 氧化应激

Abstract: BACKGROUND: Previous studies have shown that paeoniflorin has an ameliorative effect on fibrosis in liver and kidney organs, especially in hepatic fibrosis. However, the protective effect of paeoniflorin on angiotensin II-induced fibrosis in cardiac fibroblasts remains unclear.
OBJECTIVE: To investigate the protective effects of paeoniflorin on angiotensin II-induced extracellular matrix deposition in cardiac fibroblasts and molecular mechanisms.
METHODS: Angiotensin II (1 μmol/L) was added to primary isolated cultured SD rat mammary rat cardiac fibroblasts for 48 hours as the model group. The paeoniflorin low and high dose groups were pretreated with different doses of paeoniflorin (50 and 100 μmol/L) for 2 hours, and then treated with angiotensin II for 48 hours. The SIRT1 inhibitor group was treated with SIRT1 inhibitor EX527 10 μmol/L for 2 hours, followed by paeoniflorin (100 μmol/L) for 2 hours, and then co-incubation with angiotensin II for 48 hours. The cell viability was detected using the cell counting kit-8 method (CCK-8) assay. The cell migration ability was detected by Transwell. The level of intracellular reactive oxygen species was detected by DHA fluorescent probe. The level of oxidative stress markers was detected by relevant kits. Protein expression of fibrosis-related genes was detected by western blot assay. The mRNA expression levels of extracellular matrix and fibrosis-related genes were detected by qRT-PCR.
RESULTS AND CONCLUSION: (1) Compared with the control group, the proliferation and migration of cardiac fibroblasts were significantly increased after angiotensin II intervention; the intracellular content of reactive oxygen species and malondialdehyde content were elevated; the activities of superoxide dismutase and catalase were decreased, and the mRNA expression levels of α-smooth muscle actin, type I collagen, type III collagen, fibronectin, connective tissue growth factor, and matrix metalloproteinase 9 were increased. Compared with the model group, paeoniflorin could dose-dependently inhibit the above effect changes (P < 0.01). (2) Compared with the model group, paeoniflorin up-regulated the protein expression of SIRT1 in dose-dependent manner (P < 0.001). (3) Compared with the high-dose paeoniflorin group, the number of cell migration and the expression level of α-smooth muscle actin, type I collagen, and type III collagen were significantly increased in the SIRT1-inhibitor group (P < 0.01). All the experimental results show that paeoniflorin effectively attenuates the angiotensin II-induced changes in cardiac fibroblast fibrosis possibly through up-regulating the expression of SIRT1, dose-dependently inhibits cardiac fibroblast oxidative stress and extracellular matrix deposition, and has a protective effect on cardiac fibroblast fibrosis.

Key words: paeoniflorin, cardiac fibroblast, angiotensin II, extracellular matrix, fibrosis, oxidative stress

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