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 (11): 916-922.doi: 10.3969/j.issn.1006-2157.2019.11.007

• 中药药理 • 上一篇    下一篇

豁痰解毒通络方干预颈动脉粥样硬化兔的内质网应激-自噬信号通路机制研究*

于克英1, 邓悦1#, 石锐1, 赵明静2, 沈娟娟3, 高玲3, 田腾辉3, 牛望3, 王春梅3, 薛雯3, 赵越3   

  1. 1 长春中医药大学附属医院 吉林 130021;
    2 北京中医药大学东直门医院 中医内科学教育部和北京市重点实验室;
    3 长春中医药大学
  • 收稿日期:2018-11-29 出版日期:2019-11-30 发布日期:2019-12-03
  • 通讯作者: 邓悦,男,博士,主任医师,博士生导师,研究方向:中医药防治心血管病,E-mail:dyue7138@sina.com
  • 作者简介:于克英,女,博士,主治医师
  • 基金资助:
    * 国家自然科学基金面上项目(No.81774226),吉林省科技发展计划自然科学基金项目(No. 20180101149JC)

Mechanism of Huotan Jiedu Tongluo Fang in intervening endoplasmic reticulum stress-autophagy signaling pathway in rabbits with carotid atherosclerosis*

Yu Keying1, Deng Yue1#, Shi Rui2, Zhao Mingjing2, Shen Juanjuan1, Gao Ling3, Tian Tenghui1, Niu Wang3, Wang Chunmei3, Xue Wen3, Zhao Yue3   

  1. 1 Affiliated Hospital of Changchun University of Chinese Medicine, Jilin 130021, China;
    2 Education Ministry and Beijing Key Laboratory of Chinese Internal Medicine, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China;
    3 Changchun University of Chinese Medicine, Jilin 130117, China
  • Received:2018-11-29 Online:2019-11-30 Published:2019-12-03
  • Contact: Deng Yue, Ph. D., Chief Physician, Doctoral supervisor. Affiliated Hospital of Changchun University of Chinese Medicine, No. 1478, Gongnong Dalu Road, Chaoyang District, Jilin 130021. E-mail: dyue7138@sina.com

摘要: 目的 研究豁痰解毒通络方对颈动脉粥样硬化兔的内质网应激-自噬信号通路的作用。方法 10周龄雄性日本大耳兔,随机分为4组:假手术组、模型组、豁痰解毒通络方组、阿托伐他汀组。以蛋氨酸饲料喂养合球囊拉伤颈总动脉内膜法建立兔颈动脉粥样硬化模型。取损伤侧颈总动脉进行HE染色观察病理组织结构;酶联免疫吸附测定法(ELISA)检测各组血清中同型半胱氨酸(Hcy)的含量;实时荧光定量聚合酶链反应(real-time PCR)检测颈总动脉组织中的内质网应激标志分子蛋白质激酶样内质网激酶(PERK)、真核起始因子2ɑ(eIF-2ɑ)、自噬标志分子Atg5的mRNA水平;蛋白免疫印记法(Western blot)检测eIF-2ɑ、Atg5在颈总动脉组织中的蛋白表达水平。结果 与假手术组比较,模型组损伤侧颈总动脉有明显的内膜增生,血管腔变窄;血清中Hcy含量升高(P<0.01);与模型组比较,豁痰解毒通络方组损伤侧颈总动脉内膜增生明显减轻,血清中的Hcy含量减少(P<0.01)。与假手术组比较,模型组PERK、eIF-2ɑ、Atg5 mRNA水平以及eIF-2ɑ、Atg5蛋白表达水平升高(P<0.01);豁痰解毒通络方和阿托伐他汀能够降低颈动脉粥样硬化兔模型PERK、eIF-2ɑ、Atg5 mRNA水平(P<0.01)以及eIF-2ɑ、Atg5的蛋白表达水平(P<0.01)。结论 豁痰解毒通络方可以明显改善模型兔颈动脉粥样硬化情况,降低血清Hcy的含量,其机制可能是通过抑制内质网应激-自噬信号通路的过度激活实现的。

关键词: 豁痰解毒通络, 颈动脉粥样硬化, 内质网应激, 自噬,

Abstract: Objective To study the effect of Huotan Jiedu Tongluo Fang (HTJDTLF, Sweeping Phlegm-Detoxicating-Freeing Collateral Vessels Formula) on intervening endoplasmic reticulum stress (ERS)-autophagy signaling pathway in rabbits with carotid atherosclerosis (CAS). Methods Male Japanese rabbits aged 10 weeks old were randomly divided into sham-operation group, model group, HTJDTLF group and atorvastatin group. CAS model was established through feeding methionine fodder and balloon injury of common carotid artery intima. The samples of injured common carotid artery were collected and given HE staining for observing histopathological structure. The content of serum homocysteine (Hcy) was detected by using ELISA in all groups. The mRNA levels of protein kinase-like endoplasmic reticulum kinase (PERK, an ERS marker molecule), eukaryotic initiation factor 2ɑ (eIF-2ɑ) and autophagy marker molecule-Atg5 in common carotid artery tissue were detected by using real-time fluorescence quantitative polymerase chain reaction (RT-PCR). The protein expressions of eIF-2ɑ and Atg5 in common carotid artery tissue were detected by using Western blotting assay. Results Compared with sham-operation group, damaged common carotid artery had significant intimal hyperplasia and vascular lumen stanosis, and the content of serum Hcy increased (P<0.01) in model group. Compared with model group, the intimal hyperplasia was significantly relieved and content of serum Hcy decreased in HTJDTLF group (P<0.01). The mRNA levels of PERK, eIF-2ɑ and Atg5 and protein expressions of eIF-2ɑ and Atg5 increased in model group compared with sham-operation group (P<0.01). The mRNA levels of PERK, eIF-2ɑ and Atg5 decreased (P<0.01) and protein expressions of eIF-2ɑ and Atg5 decreased (P<0.01) in HTJDTLF group and atorvastatin group. Conclusion HTJDTLF can significantly relieve CAS and reduce serum Hcy content in rabbits with CAS, and the mechanism maybe related to the inhibition of excessive activation of ERS-autophagy signaling pathway

Key words: sweeping phlegm, detoxicating and freeing collateral vessels, carotid atherosclerosis, endoplasmic reticulum stress, autophagy, rabbits

中图分类号: 

  • R285.5