南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (5): 694-701.doi: 10.12122/j.issn.1673-4254.2023.05.03

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左归降糖解郁方促进糖尿病并发抑郁症大鼠的海马齿状回神经干细胞的自我更新并激活Shh信号通路

杨 蕙，王 华，李成龙，何 雄，雷诗卉，李 薇，孟 盼，王瑾茜，刘 检，王宇红   

  1. 湖南中医药大学第一附属医院医学创新中心，湖南 长沙 410007；科技创新中心，湖南 长沙 410208；中国人民解放军联勤保障部队第九二一医院急诊科，湖南 长沙 410153；湖南省药品审评与不良反监测中心，湖南 长沙 410013
  • 出版日期:2023-05-20 发布日期:2023-06-12

Zuogui Jiangtang Jieyu Decoction promotes neural stem cell self-renewal and activates Shh signaling in the hippocampal dentate gyrus of diabetic rats with depression

YANG Hui, WANG Hua, LI Chenglong, HE Xiong, LEI Shihui, LI Wei, MENG Pan, WANG Jinxi, LIU Jian, WANG Yuhong   

  1. Centre for Medical Innovations, First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410007, China; Science and Technology Innovation Center, Changsha, 410208, China; Emergency Department, 921th Hospital of Joint Logistics Support Force of the Chinese People's Liberation Army, Changsha 410153, China; Hunan Provincial Drug Evaluation and Adverse Reaction Monitoring Center, Changsha 410013, China
  • Online:2023-05-20 Published:2023-06-12

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摘要： 目的 研究左归降糖解郁方调节糖尿病并发抑郁症大鼠海马Shh信号通路，改善齿状回神经干细胞自我更新的机制。方法 建立糖尿病并发抑郁症大鼠模型并随机分为5组，16只/组：模型组、阳性药（二甲双胍+氟西汀）组、左归降糖解郁方高、中、低剂量 组；另设健康SD大鼠为正常组。其中，左归降糖解郁方高、中、低剂量组和阳性药组分别给予相应药物，对照组和模型组灌胃给予 等体积蒸馏水。给药结束后，采用血糖试纸检测大鼠血糖，采用强迫游泳实验和水迷宫实验检测大鼠抑郁样行为和学习记忆功 能，采用酶联免疫吸附法检测大鼠血清瘦素水平。采用双重免疫荧光法检测海马齿状回Nestin和Brdu蛋白表达；采用Western blot法检测海马神经干细胞自我更新标（P<0.01）志物及Shh信号蛋白的表达。结果 与正常组比较，模型组大鼠血糖和瘦素水平 显著升高（P<0.01），强迫游泳实验的不动时间显著延长（P<0.01），水迷宫实验中动物登台时间明显延长，寻台时间明显缩短，穿台 次数明显减少（P<0.01）；海马齿状回Nestin和Brdu表达显著减少（P<0.01），Cyclin D1、SOX2、Shh、Ptch1、Smo蛋白表达显著降低 （P<0.01），Gli-3表达显著升高（P<0.01），细胞核内Gli-1蛋白表达显著减少（P<0.01）。与模型组比较，左归降糖解郁方高剂量可显 著降低模型大鼠的血糖（P<0.01）和瘦素水平（P<0.05），缩短动物在强迫游泳实验中的不动时间（P<0.01）和在水迷宫实验中的登 台时间（P<0.05），延长寻台时间并增加穿台次数（P<0.01）。同时，该方高剂量可显著增加齿状回Nestin和Brdu表达（P<0.01），增 加海马Cyclin D1、SOX2、Shh、Ptch1、Smo蛋白表达（P<0.01），降低Gli-3表达（P<0.05），并增加Gli-1蛋白的核表达（P<0.01）。结 论 左归降糖解郁方可有效改善糖尿病并发抑郁症大鼠海马齿状回神经干细胞的自我更新能力，并激活干细胞Shh信号通路。

关键词: 左归降糖解郁方；糖尿病并发抑郁症；神经干细胞；自我更新；Shh

Abstract: Objective To investigate the effect of Zuogui Jiangtang Jieyu Decoction (ZJJ) on Shh signaling and self-renewal of neural stem cells in the hippocampal dentate gyrus of diabetic rats with depression. Methods Diabetic rat models with depression were randomly divided into model group, positive drug (metformin+ fluoxetine) group, and low-, medium-, and high-dose ZJJ groups (n=16), with normal SD rats as the control group. The positive drugs and ZJJ were administered by gavage, and the rats in the control and model groups were given distilled water. After the treatment, blood glucose level was detected using test strips, and behavioral changes of the rats were assessed by forced swimming test and water maze test. ELISA was used to examine the serum level of leptin; The expressions of nestin and Brdu proteins in the dentate gyrus of the rats were detected using immunofluorescence assay, and the expressions of self-renewal marker proteins and Shh signaling proteins were detected using Western blotting. Results The diabetic rats with depression showed significantly increased levels of blood glucose and leptin (P<0.01) and prolonged immobility time in forced swimming test (P<0.01) and increased stage climbing time with reduced stage seeking time and stage crossings in water maze test (P<0.01). The expressions of nestin and Brdu in the dentate gyrus, the expressions of cyclin D1, SOX2, Shh, Ptch1, Smo in the hippocampus and the nuclear expression of Gli-1 were decreased (P<0.01) while hippocampal Gli-3 expression was increased significantly (P<0.01) in the rat models. Treatment of rat models with high-dose ZJJ significantly reduced the blood glucose (P<0.01) and leptin level (P<0.05) and improved their performance in behavioral tests (P<0.01). The treatment also obviously increased the expressions of nestin, Brdu, cyclin D1, SOX2, Shh, Ptch1, and Smo and the nuclear expression of Gli-1 in the dentate gyrus (P<0.01) and reduced hippocampal expression of Gli-3 (P<0.05) in the rat models. Conclusion ZJJ can significantly improve the self-renewal ability of neural stem cells and activate Shh signaling in dentate gyrus of diabetic rats with depression.

Key words: Zuogui Jiangtang Jieyu Decoction; diabetes complicated with depression; neural stem cells; self-renewal; Shh signaling