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 (5): 396-403.doi: 10.3969/j.issn.1006-2157.2019.05.007

• 中药药理 • 上一篇    下一篇

左归降糖解郁方对糖尿病并发抑郁症大鼠海马NVU连接蛋白表达的影响*

黄会珍1, 柳卓1, 韩远山2, 赵洪庆1,3, 金狮1, 罗薇絮1, 孟盼1,3, 王宇红1,3#   

  1. 1 湖南中医药大学中药粉体与创新药物省部共建国家重点实验室培育基地 湖南 410208;
    2 湖南中医药大学第一附属医院;
    3 湖南中医药大学科技创新中心
  • 收稿日期:2018-11-06 出版日期:2019-05-30 发布日期:2019-06-05
  • 通讯作者: 王宇红, 女, 博士, 研究员, 博士生导师, 研究方向:中药神经药理, E-mail: wyh107@126.com
  • 作者简介:黄会珍,女,在读硕士生
  • 基金资助:
    *国家科技重大专项重大新药创制项目(No.2017ZX09309026),国家自然科学基金资助项目(No.81573965、No.81874464),国家自然科学基金青年基金(No.81703694),“湖南中医药大学中医学国内一流建设学科”项目

Influence of Zuogui Jiangtang Jieyu Fang on expressions of hippocampal NVU connexins in rats with diabetes mellitus complicated by depression*

Huang Huizhen1, Liu Zhuo1, Han Yuanshan2, Zhao Hongqing1,3, Jin Shi1, Luo Weixu1, Meng Pan1,3, Wang Yuhong1,3#   

  1. 1 Training Base of Province-Ministry Joint State Key Laboratory of Chinese Medicinal Powder and Innovative Medicinals, Hunan University of Chinese Medicine, Hunan 410208, China;
    2 First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410007, China;
    3 Science and Technology Innovation Center, Hunan University of Traditional Chinese Medicine, Hunan 410208, China
  • Received:2018-11-06 Online:2019-05-30 Published:2019-06-05
  • Contact: Wang Yuhong, Ph. D., Researcher, Doctoral Supervisor. Training Base of Province-Ministry Joint State Key Laboratory of Chinese Medicinal Powder and Innovative Medicinals, Science and Technology Innovation Center, Hunan University of Chinese Medicine, No. 300, Xueshi Road, Hanpu Science and Education Park, Yuelu District, Changsha 410208. E-mail: wyh107@126.com

摘要: 目的 基于神经血管单元(NVU)探讨左归降糖解郁方对糖尿病并发抑郁症大鼠海马紧密和缝隙连接蛋白表达的影响。方法 70只成年雄性SD大鼠高脂乳剂灌胃、尾静脉注射链脲佐菌素联合慢性不可预见性应激建立糖尿病并发抑郁症动物模型,并随机分为5组:模型组,二甲双胍+氟西汀组,左归降糖解郁方高、中、低剂量组,另取12只未造模大鼠作为空白对照组,于应激造模同时灌胃给药28 d。旷场实验和Morris水迷宫评价大鼠自主活动水平和学习记忆能力,透射电镜观察大鼠海马NVU形态结构,免疫组化和实时荧光定量PCR检测大鼠海马紧密连接蛋白Claudin-1、Occludin和缝隙连接蛋白43(CX43)、水通道蛋白4(AQP4)的表达量。结果 与空白对照组相比,模型组大鼠自主活动次数和空间探索时间明显减少、逃避潜伏期明显延长(P<0.01);透射电镜和病理结果显示模型组大鼠海马内皮细胞肿胀,血管周围紧密连接和缝隙连接紊乱,细胞排列松散,Claudin-1、Occludin、CX43和AQP4及其mRNA表达显著降低(P<0.01)。与模型组相比,左归降糖解郁方高剂量组自主活动次数和空间探索时间显著增加、逃避潜伏期显著减少(P<0.05、 P<0.01);海马区域细胞水肿减轻,血管周围紧密和缝隙连接恢复,细胞排列较为整齐均一,Claudin-1、Occludin、CX43和AQP4及其mRNA表达明显升高(P<0.05、 P<0.01)。结论 左归降糖解郁方能明显缓解大鼠抑郁样行为,其机制可能与增强大鼠海马紧密和缝隙连接蛋白表达,维持海马NVU整体结构与功能有关。

关键词: 左归降糖解郁方, 糖尿病并发抑郁症, 神经血管单元, 紧密连接蛋白, 缝隙连接蛋白, 大鼠

Abstract: Objective To discuss the influence of Zuogui Jiangtang Jieyu Fang (ZHJYF, Left-Restoring Sugar-lowering Depression-relieving Formula), based on neurovascular unit (NVU), on the expressions of tight junction proteins and connexins in rats with diabetes mellitus (DM) complicated by depression. Methods Adult SD rats (n=70) were orally given high-fat emulsion, injection of streptozocin (STZ) in caudal vein and chronic unpredicted stress to establish the model of DM complicated by depression. The rats were randomly divided into 5 groups: model group, metformin+fluoxetine group, high-dose, mid-dose and low-dose ZHJYF groups. Other 12 unmodeling rats were chosen into blank control group. All group were orally given corresponding medicinals at the same time of stress for 28 d. The level of autonomic activity and learning-memory ability were reviewed by using open-field test and Morris water maze (MWM), and morphologic structure of hippocampal NVU was observed by using transmission electron microscope (TEM). The expressions of tight junction proteins (Claudin-1 and Occluding), connexins [aonnexin43 (CX43) and aquaporin4 (AQP4)] were detected by using immunohistochemistry technique and RT-qPCR. Results Compared with blank control group, the times of autonomic activity and space exploration time decreased significantly and escape latency was prolonged significantly in model group (P<0.01). The results of TEM and pathology showed that swollen hippocampal endothelial cells, confused perivascular tight and gap junctions and loosely arranged cells, and the protein and mRNA expressions of Claudin-1, Occludin, CX43 and AQP4 decreased significantly in model group (P<0.01). Compared with mode group, the times of autonomic activity and space exploration time increased significantly and escape latency was shortened significantly (P<0.05 or P<0.01), swelling of hippocampal endothelial cells was relieved, perivascular tight and gap junctions was recovered, and cell arrangement was tidiness, and the protein and mRNA expressions of Claudin-1, Occludin, CX43 and AQP4 increased significantly in high-dose ZHJYF group (P<0.05 or P<0.01). Conclusion ZHJYF can significantly relieve depressive behavior in rats, and the mechanism may be related to enhance of expressions of tight junction proteins and connexins and maintain the structure and function of hippocampal NVU.

Key words: Zuogui Jiangtang Jieyu Fang (Left-Restoring Sugar-lowering Depression-relieving Formula), diabetes mellitus complicated by depression, hippocampal neurovascular unit, tight junction proteins, connexins, rats

中图分类号: 

  • R285.5