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 ›› 2021, Vol. 44 ›› Issue (9): 846-853.doi: 10.3969/j.issn.1006-2157.2021.09.013

• 中药药理 • 上一篇    下一篇

基于PERK-eIF2α信号通路探讨冠心康及其拆方对高脂血症的调控机制*

马贵萍1,2,3, 于忠杨4, 孙乐1,2,3, 卿立金1, 黄育生1,2,3, 章怡祎5, 洪创雄1,2#   

  1. 1 广州中医药大学第一附属医院 广东 510405;
    2 广州中医药大学第一临床医学院;
    3 广州中医药大学岭南医学研究中心;
    4 青岛市第六人民医院;
    5 上海中医药大学附属龙华医院
  • 收稿日期:2021-03-12 出版日期:2021-09-30 发布日期:2021-10-08
  • 通讯作者: #洪创雄,男,硕士,教授,博士生导师,主要研究方向:中医药防治心血管疾病,E-mail:hongchuangxiong@gzucm.edu.cn
  • 作者简介:马贵萍,女,在读博士生
  • 基金资助:
    *国家自然科学基金项目(No.81373799),广东省中医药强省建设指标体系构建研究项目(No.20173004)

Study on the mechanism of the regulatory effects of Guanxinkang and its composite formulas on hyperlipidemia based on PERK-eIF2α signaling pathway*

Ma Guiping1,2,3, Yu Zhongyang4, Sun Le1,2,3, Qing Lijin1, Huang Yusheng1,2,3, Zhang Yiyi5, Hong Chuangxiong1,2#   

  1. 1 The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangdong 510405, China;
    2 First School of Clinical Medicine of Guangzhou University of Chinese Medicine, Guangdong 510405, China;
    3 Lingnan Medical Research Center, Guangzhou University of Chinese Medicine, Guangdong 510405, China;
    4 Qingdao No.6 People's Hospital, Shandong 266033, China;
    5 Long Hua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
  • Received:2021-03-12 Online:2021-09-30 Published:2021-10-08
  • Contact: Associate Prof. Hong Chuangxiong, MA, Doctoral Supervisor. The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine.No.12 Airport Road, Baiyun District, Guangzhou 510405.E-mail: hongchuangxiong@gzucm.edu.cn
  • Supported by:
    National Natural Science Foundation of China (No. 81373799); Research on the Construction of the Index System for Building a Strong Province of Traditional Chinese Medicine in Guangdong Pro-vince (No. 20173004)

摘要: 目的 探讨冠心康及其拆方对肝脏胆固醇逆转运的调节及机制。方法 60只低密度脂蛋白受体基因敲除(LDLR-/-)小鼠,雌雄各半,高脂饮食诱导12周,建立高脂血症模型。对模型验证后,按照高脂血症的临床常见证型将冠心康全方分为3个拆方组(益气化痰组、化痰活血组、益气活血组),60只LDLR-/-小鼠随机分为模型组、冠心康组、益气化痰组、化痰活血组、益气活血组和辛伐他汀组,每组10只,另将10只相同遗传背景C57BL/6J小鼠设定为正常组。按照分组,分别给予冠心康及其拆方以及辛伐他汀干预,12周后同时取小鼠外周血及肝脏。生化法检测各组小鼠血脂含量;HE染色观察肝脏病理形态变化;实时荧光定量(RT-PCR)法检测肝脏蛋白激酶R样内质网激酶(PERK)、真核翻译起始因子2α(eIF2α)、甾醇调节元件结合蛋白-2(SREBP-2)、ATP结合盒转运子亚家族成员1(ABCA1)基因表达;免疫印迹法(Western blotting)检测肝脏磷酸化PERK、磷酸化eIF2α、ABCA1和SREBP-2蛋白表达。结果 冠心康全方及其拆方对血脂有不同程度的调节作用;冠心康组、益气化痰组小鼠肝组织结构较完整,化痰活血组、益气活血组小鼠肝脏视野中可见大量的脂滴空泡,伴有明显疏松呈网状改变的胞质;与模型组相比,冠心康组小鼠肝脏PERK、eIF2α、SREBP-2基因和蛋白表达明显下降,ABCA1蛋白明显升高(P<0.05),3个拆方组则差异无统计学意义(P>0.05)。结论 冠心康可有效调节血脂,维持肝脏胆固醇稳态;冠心康对血脂的调节和肝脏胆固醇的“维稳”作用,可能与其通过激活ABCA1和抑制PERK-eIF2α-SREBP-2信号通路,从而减少胆固醇的合成、摄取和促进胆固醇的转运、代谢有关;并且,冠心康中益气活血的药物(黄芪、益母草、丹参)对PERK-eIF2α-SREBP-2信号通路的抑制作用更为明显。

关键词: 高脂血症, 冠心康, 内质网应激, 胆固醇稳态, 小鼠

Abstract: Objective To explore the regulatory effects of Guanxinkang (a patent Chinese medicine based on the Trichosanthes, Chinese Chive and Pinellia Decoction) and its composite formulas on liver cholesterol reverse transport and its mechanism.Methods 60 LDLR-/- mice, half male and half female, were induced by high-fat diet for 12 weeks to establish the hyperlipidemia model. After verifying the model, three composite formulas were obtained from the complete formula of Guangxinfang according to the commonly encountered clinical patterns of hyperlipidemia, i.e. Yiqi Huatan (qi-reinforcing and phlegm-resolving, YQHT)formula, Huatan Huoxue (phlegm-resolving and blood-activating, HTHX) formula, and Yiqi Huoxue (qi-reinforcing and blood-activating, YQHX) formula. The 60 mice were randomly divided into the model group, Guanxinkang (GXK) group, Yiqi Huatan (YQHT) group, Huatan Huoxue (HTHX) group, Yiqi Huoxue (YQHX) group, and simvastatin group, 10 mice in each group, and another 10 C57BL/6J mice with the same genetic background formed the normal group.Guanxinkang, its composite formulas and simvastatin were given to the corresponding groups respectively. Peripheral blood and liver of the mice were collected 12 weeks later. Biochemical method was used to detect the blood lipid content of mice in different groups, HE staining to observe liver pathological changes,real-time PCR to detect liver PERK, eIF2α, SREBP-2, ABCA1 gene expressions, and Western Blotting to detect liver phosphorylation PERK, phosphorylation eIF2α, ABCA1 and SREBP-2 protein expressions. Results Guanxinkang and its composite formulas can regulate blood lipids to different extents. The liver tissues of the mice in the GXK group and the YQHT group were relatively complete. In the liver of the mice in the HTHX group and YQHX group, a large number of lipid droplet vacuoles were seen, accompanied by obvious loose cells with network-like changes. Compared with the model group, the liver PERK, eIF2α, SREBP-2 gene and protein expressions of mice in the GXK group were significantly reduced, and the ABCA1 protein was significantly increased(P<0.05), but there was no such statistically significant difference in the three groups taking the composite formulas (P>0.05). Conclusion Guanxinkang can effectively regulate blood lipids and maintain liver cholesterol homeostasis. Guanxinkang's regulation of blood lipids and its“stabilizing” effects on liver cholesterol may be related to the activation of ABCA1 and inhibition of PERK-eIF2α-SREBP-2 signaling pathway, which reduces the synthesis and uptake of cholesterol and promotes the transport and metabolism of cholesterol. In addition, the herbal ingredients of Guanxinkang that invigorate qi and activate blood, including Huangqi (Astragalus Root, Radix Astragali), Yimucao (Motherwort Herb, Herba Leonuri), and Danshen (Danshen Root, Radix et Rhizoma Salviae Miltiorrhizae) inhibited the PERK-eIF2α-SREBP-2 signaling pathway more significantly.

Key words: hyperlipidemia, Guanxinkang, endothelial reticulum stress, cholesterol homeostasis, mouse

中图分类号: 

  • R285.5