左归降糖解郁方改善糖尿病并发抑郁症大鼠N-甲基D-天冬氨酸受体过激致海马突触损伤的作用机制  

Mechanism of Zuogui Jiangtang Jieyu Formula on improving hippocampal synaptic damage caused by N-methyl D-aspartate receptor hyperactivity in diabetes with depression rats

作　　者：刘洋[1] 李萍[1] 杨蕙[3] 邹蔓姝 赵洪庆[1] 郭海鹏 王婷婷 王华[3] 王宇红[1,2] LIU Yang;LI Ping;YANG Hui;ZOU Man-shu;ZHAO Hong-qing;GUO Hai-peng;WANG Ting-ting;WANG Hua;WANG Yu-hong(Huan University of Chinese Medicine,Changsha 410208,China;Hunan Key Laboratory of Traditional Chinese Medicine Prevention&Treatment of Depressive Diseases,Changsha 410208,China;First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]抑郁类疾病中医药防治湖南省重点实验室,湖南长沙410208 [3]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《中草药》2023年第19期6323-6335,共13页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81874464);国家自然科学基金资助项目(82174357);湖南省科药联合基金资助项目(2023JJ60476);湖南省科药联合基金资助项目(2023JJ60482);湖南省中医药管理局科研课题(B2023141)。

摘　　要：目的研究左归降糖解郁方通过调节吲哚胺-2,3-双加氧酶(indoleamine 2,3-dioxygenasel,IDO)信号抑制糖尿病并发抑郁症大鼠海马N-甲基D-天冬氨酸受体(N-methyl D-aspartate receptor,NR)过激致海马突触损伤的作用机制。方法建立糖尿病并发抑郁症大鼠模型,并随机分为空白组、模型组、IDO抑制剂(3 mg/kg)组、左归降糖解郁方(20.52 g/kg)组和阳性对照(二甲双胍0.18 g/kg+氟西汀1.8 mg/kg)组。各组大鼠ig给药28 d后,采用旷场实验、强迫游泳实验和Morris水迷宫检测大鼠抑郁样行为和记忆认知功能;采用Western blotting和qRT-PCR检测海马IDO蛋白和基因表达;采用ELISA试剂盒检测血清和海马中γ-干扰素(interferon-γ,IFN-γ)、CC族趋化因子配体2(CC chemokine ligand 2,CCL2)、色氨酸、5-羟色胺(5-hydroxytryptamine,5-HT)、犬尿氨酸、犬尿喹啉酸(kynurenine acid,KA)、喹啉酸、白细胞介素-1β(interleukin1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和IL-6水平;采用免疫荧光法检测海马中离子钙接头蛋白-1(ionized calcium binding adaptor molecule-1,Iba-1)、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、突触素(synaptophysin,Syn)、突触后致密蛋白-95(post synaptic density protein-95,PSD-95)的表达;采用Western blotting检测海马中腺苷A1R受体(adenosine A1 receptor,A1R)、腺苷A2AR受体(adenosine A2A receptor,A2AR)、腺苷A3R受体(adenosine A3 receptor,A3R)、N-甲基D-天冬氨酸受体2A(N-methyl D-aspartate receptor 2A,NR2A)、N-甲基D-天冬氨酸受体2B(N-methyl D-aspartate receptor 2B,NR2B)和脑源性神经营养因子(brain derived neurotrophic factor,BDNF)蛋白表达;采用苏木素-伊红染色、Nissl染色和TUNEL法检测海马病理损伤及凋亡情况。结果左归降糖解郁方能显著降低模型大鼠血糖水平(P<0.01),并改善抑郁样行为,表现为旷场实验总距离和中心时间显著增加(P<0.01)、强迫游泳不动时间显著缩短(P<0.01)、Morris水迷�Objective To study the mechanism of Zuogui Jiangtang Jieyu Formula(左归降糖解郁方)on inhibiting hippocampal synaptic damage caused by excessive N-methyl D-aspartate receptor(NR)in diabetes with depression rats by regulating indoleamine 2,3-dioxygenasel(IDO)signal.Method Diabetic rats with depression were established and randomly divided into blank group,model group,IDO inhibitor(3 mg/kg)group,Zuogui Jiangtang Jieyu Formula(20.52 g/kg)group and positive control(metformin 0.18 g/kg+fluoxetine 1.8 mg/kg)group.After 28 d of administration,depression-like behavior and memory and cognitive function were detected by open field test,forced swimming test and Morris water maze.Western blotting and qRT-PCR were used to detect the expression levels of IDO protein and mRNA in hippocampus.The contents of interferon-γ(IFN-γ),CC chemokine ligand 2(CCL2),tryptophan,5-hydroxytryptamine(5-HT),kynurenine,kynurenine acid(KA),quinolinic acid,interleukin-1β(IL-1β),tumor necrosis factor-α(TNFα)and IL-6 in serum and hippocampus were determined by ELISA.The expressions of ionized calcium binding adaptor molecule-1(Iba-1),glial fibrillary acidic protein(GFAP),synaptophysin(Syn)and post synaptic density protein-95(PSD-95)in hippocampus were detected by immunofluorescence assay.Protein expressions of adenosine A1 receptor(A1R),adenosine A2AR receptor(A2AR),adenosine A3 receptor(A3R),N-methyl D-aspartate receptor 2A(NR2A),N-methyl D-aspartate receptor 2B(NR2B)and brain derived neurotrophic factor(BDNF)in hippocampus were detected by Western blotting.Hematoxylin-eosin(HE),Nissl and TUNEL were used to detect the pathological injury and apoptosis of hippocampus.Results Zuogui Jiangtang Jieyu Formula significantly reduced blood sugar level and improved depression-like behavior in model rats(P<0.01).The results showed that the total distance and center time of open field experiment were significantly increased(P<0.01),the immobile time of forced swimming was significantly shortened(P<0.01),the positioning cruise time of Morris water m

关 键 词：左归降糖解郁方 糖尿病并发抑郁症 吲哚胺-2 3-双加氧酶 色氨酸-犬尿氨酸代谢通路 海马突触损伤 马钱苷 芍药苷 毛蕊异黄酮苷 金丝桃苷 姜黄素 丹酚酸B 

分 类 号：R285.5[医药卫生—中药学]