主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2019, Vol. 42 ›› Issue (8): 655-661.doi: 10.3969/j.issn.1006-2157.2019.08.008

• 中药药理 • 上一篇    下一篇

丹参通络解毒汤联合骨髓干细胞移植对急性心肌梗死大鼠心肌纤维化的影响及其机制*

李鑫辉; 黄淼鑫; 杜建芳; 陈聪; 杨艳红; 吴若霞; 王静雯; 李彩云   

  1. 湖南中医药大学中医学院 湖南 410208
  • 收稿日期:2019-02-15 出版日期:2019-08-30 发布日期:2019-09-04
  • 作者简介:李鑫辉,男,博士,教授,硕士生导师
  • 基金资助:
    *国家自然科学基金资助项目(No.81704065),湖南省自然科学基金资助项目(No.2019JJ40210),湖南省教育厅科学研究重点资助项目(No.15A143),湖南省中医药科研计划资助项目(No.201559,No.201790),长沙市科技计划资助项目(No.kq1706058)

Influence and mechanism of Danshen Tongluo Jiedu Tang combined with bone marrow stem cell transplantation on myocardial fibrosis in rats with acute myocardial infarction*

Li Xinhui, Huang Miaoxin, Du Jianfang, Chen Cong, Yang Yanhong, Wu Ruoxia, Wang Jingwen, Li Caiyun   

  1. School of Chinese Medicine, Hunan University of Chinese Medicine, Hunan 410208, China
  • Received:2019-02-15 Online:2019-08-30 Published:2019-09-04
  • Contact: Pro. Li Xinhui, Ph. D., Master’s supervisor. School of Chinese Medicine, Hunan University of Chinese Medicine, No. 300, Xueshi Road, Hanpu Science and Education Park, Yuelu District, Changsha 410208. E-mail: 3077314414@qq.com

摘要: 目的 探讨丹参通络解毒汤联合骨髓干细胞移植对急性心肌梗死(AMI)大鼠心肌纤维化的影响及作用机制。方法 采用全骨髓贴壁法结合密度梯度离心法分离和培养骨髓间充质干细胞(BMSCs)。经结扎冠状动脉的左前降支(LAD) 建立心梗模型。大鼠随机分为假手术组、AMI模型组、BMSCs移植组、中药组(丹参通络解毒汤组)、联合组(丹参通络解毒汤+ BMSCs移植组)。除假手术组外其余组造模,术后中药组与联合组分别灌胃丹参通络解毒汤,其余各组用等量生理盐水灌胃,1次/d,连续14 d。Masson染色观察心肌组织胶原含量,ELISA法测定血清Ⅲ型前胶原末端肽(PⅢP)、Ⅳ型前胶原末端肽(PⅣP)、一氧化氮(NO)、内皮素-1(ET-1)浓度。蛋白印迹法检测心肌组织中内皮细胞蛋白c受体(EPCR)和血栓调节蛋白(TM)的表达水平。采用SPSS 19.0 统计软件对数据进行分析。结果 与假手术组比较,各组大鼠EPCR及TM蛋白表达下降,NO水平降低,ET-1、PⅢP、PⅣP水平升高,心肌有不同程度的胶原纤维产生,差异有显著性(P<0.01);与AMI模型组比较,BMSCs移植组、中药组和联合组EPCR和TM蛋白表达及NO水平均显著上升,ET-1、PⅢP及PⅣP水平均降低,胶原纤维产生均减少,差异具有显著性(P<0.01);与BMSCs移植组比较,中药组EPCR和TM蛋白表达及NO水平均上升,ET-1、PⅢP及PⅣP水平均降低,胶原纤维产生均减少,差异具有显著性(P<0.01);与中药组比较,联合组EPCR和TM蛋白表达及NO水平均上升,ET-1、PⅢP及PⅣP水平均降低,胶原纤维产生均减少,差异具有显著性(P<0.01)。结论 丹参通络解毒汤联合BMSCs移植能够抑制心肌纤维化产生,其机制可能与上调AMI大鼠心肌EPCR及TM蛋白表达,保护血管内皮功能有关。

关键词: 急性心肌梗死, 内皮细胞蛋白c受体, 血栓调节蛋白, 骨髓干细胞, 丹参通络解毒汤, Ⅲ型前胶原末端肽, Ⅳ型前胶原末端肽, 大鼠

Abstract: Objective To investigate the influence and mechanism of Danshen Tongluo Jiedu Tang (Salvia Root Collateral-freeing Removing Toxicity Decoction) combined with bone marrow stem cell transplantation (BMSCT) on myocardial fibrosis in rats with acute myocardial infarction (AMI). Methods The bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured by using whole bone marrow adherence method combined with density gradient centrifugation. The model of AMI was established through ligating the left anterior descending branch (LAD) of coronary artery. The rats were randomly divided into sham-operation group, AMI model group, BMSCT group, Danshen Tongluo Jiedu Tang group (Chinese medicinal group) and Danshen Tongluo Jiedu Tang+BMSCT group (combination group). The model was established in all groups except of sham-operation group, and then Chinese medicinal group and combination group were respectively given orally DTJT and other groups were given orally equivalent saline water once a day for 14 days continuously. The content of collagen in myocardial tissue was observed after Masson staining, and concentration of procollagen type III N-terminal peptide (PⅢP), procollagen type IV N-terminal peptide (PIVP), nitric oxide (NO) and endothelin-1 (ET-1) were detected by using ELISA. The expressions of endothelial protein C receptor (EPCR) and thrombomodulin (TM) were detected by using Western blotting assay, and all data was analyzed by using SPSS 19.0 statistical software. Results Compared with sham-operation group, the protein expressions of EPCR and TM decreased, NO level decreased, levels of ET-1, PⅢP and PⅣP increased, and myocardial collagen fiber generation in varying degrees was observed in other groups (P<0.01). The protein expressions of EPCR and TM and NO level increased, and levels of ET-1, PⅢP and PⅣP and myocardial collagen fiber generation decreased in BMSCT group, Chinese medicinal group and combination group compared with AMI group (P<0.01). The protein expressions of EPCR and TM and NO level increased, and levels of ET-1, PⅢP and PⅣP and myocardial collagen fiber generation decreased in Chinese medicinal group compared with BMSCT group (P<0.01). The protein expressions of EPCR and TM and NO level increased, and levels of ET-1, PⅢP and PⅣP and myocardial collagen fiber generation decreased in combination group compared with Chinese medicinal group (P<0.01). Conclusion Danshen Tongluo Jiedu Tang combined with BMSCT can inhibit myocardial fibrosis, and the mechanism may be related to that the protein expressions of myocardial EPCR and TM are up-regulated and vascular endothelial function is protected in AMI rats.

Key words: acute myocardial infarction, endothelial protein C receptor, thrombomodulin, bone marrow stem cells, Danshen Tongluo Jiedu Tang (Salvia Root Collateral-freeing Removing Toxicity Decoction), procollagen type III N-terminal peptide, procollagen type IV N-terminal peptide, rats

中图分类号: 

  • R285.5