左归降糖解郁方调控CD300f/GLUT1信号通路改善糖尿病并发抑郁症大鼠海马神经元突触损伤的作用机制  

Zuogui Jiangtang Jieyu Formula regulates the CD300f/GLUT1 signaling pathway to improve the synaptic damage of hippocampal neurons in rats with diabetes-related depression

作　　者：刘检[1] 唐林[1] 赵洪庆[2] 姜帆[1] 刘林[1] 胡超 LIU Jian;TANG Lin;ZHAO Hongqing;JIANG Fan;LIU Lin;HU Chao(The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Science&Technology Innovation Center,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学第一附属医院,长沙410007 [2]湖南中医药大学科技创新中心

出　　处：《北京中医药大学学报》2024年第11期1573-1584,共12页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金青年项目(No.82104793);湖南省自然科学基金面上项目(No.2022JJ30451);湖南省三尖创新人才工程湖湘青年英才项目(No.2022RC1226)。

摘　　要：目的基于白细胞单免疫球蛋白样受体3(CD300f)/葡萄糖转运体1(GLUT1)信号通路介导小胶质细胞糖代谢,探讨左归降糖解郁方改善糖尿病并发抑郁症大鼠海马神经元突触损伤的作用机制。方法80只雄性SD大鼠按照随机数字表法选取10只大鼠作为正常组,其余70只高脂饲料喂养4周后单次尾静脉注射链脲佐菌素(38 mg/kg)复制大鼠糖尿病模型,筛选造模成功的大鼠60只,按随机数字表法分为模型组、CD300f阻断剂组、CD300f激动剂组、二甲双胍+氟西汀组(二甲双胍0.18 g/kg+氟西汀1.8 mg/kg)和左归降糖解郁方高、低剂量组(20.52、10.26 g/kg)。除正常组外,其余各组大鼠予28 d慢性不可预知温和应激加孤养复制糖尿病并发抑郁症大鼠模型。其中,二甲双胍+氟西汀组与左归降糖解郁方高、低剂量组在造模第2周后连续灌胃相应药物,正常组、模型组灌胃等体积蒸馏水,连续14 d;CD300f阻断剂组、激动剂组则进行海马区微量注射给药,分别注射髓系细胞触发受体抑制因子(CLM1,2μg/kg)和免疫球蛋白Fc段表面蛋白(Fcγ,5μg/kg),每周1次。干预结束后,采用旷场实验、强迫游泳实验检测大鼠抑郁样行为;生化分析检测大鼠海马组织葡萄糖、乳酸含量及腺苷二磷酸(ADP)/三磷酸腺苷(ATP);采用酶联免疫吸附测定法检测大鼠海马组织胰岛素、5-羟色胺(5-HT)和多巴胺(DA)水平;采用免疫荧光检测海马组织CD300f、GLUT1、突触相关前膜蛋白3(RIMS3)、突触后膜相关蛋白102(SAP102)平均荧光强度;蛋白质印迹法检测大鼠海马组织CD300f、GLUT1、RIMS3、SAP102蛋白表达情况;采用尼氏染色和透射电镜观察大鼠海马组织病理改变。结果与正常组比较,模型组大鼠旷场总活动路程减少、强迫游泳不动时间增加,海马组织葡萄糖、乳酸含量及ADP/ATP升高、胰岛素水平降低、5-HT和DA水平下降,海马组织CD300f、GLUT1、RIMS3、SAP102平均荧光强度及蛋白相�Objective To explore the protective mechanism of Zuogui Jiangtang Jieyu Formula(ZGF)on synaptic damage of hippocampal neurons based on leukocyte mono-immunoglobulin-like receptor 3(CD300f)/glucose transporter 1(GLUT1)signal-mediated microglial glucose metabolism in rats with diabetes-related depression.Methods Eighty male SD rats were randomly selected using random number table method,with 10 rats serving as the normal group.The remaining 70 rats were fed a high-fat diet for 4 weeks and then injected once with 38 mg/kg of streptozotocin via the tail vein to replicate the diabetes rat model.Sixty rats were screened and successfully modeled,which were randomly divided into the model,CD300f blocker,CD300f agonist,metformin+fluoxetine(metformin 0.18 g/kg+fluoxetine 1.8 mg/kg),and ZGF high-and low-dose(20.52 and 10.26 g/kg,respectively)groups using random number table method.In addition to the normal group,the rats in the other groups underwent chronic unpredictable mild stress combined with isolation feeding for 28 days to replicate the diabetes-related depression rat model.The metformin+fluoxetine and ZGF high-and low-dose groups were subjected to continuous intragastrial administration for 14 days after the second week of modeling.The normal and model groups were administered an equal amount of distilled water by gavage.The CD300f blocker group and agonist group received microinjection into the hippocampus,with injection of myeloid cell trigger receptor inhibitory factor(CLM1,2μg/kg)and immunoglobulin Fc surface protein(Fcγ,5μg/kg)once a week,respectively.Depression-like behavior in rats was evaluated using open-field and forced swimming tests after the intervention.Biochemical analysis was used to detect the glucose,lactic acid,and adenosine diphosphate(ADP)/adenosine triphosphate(ATP)ratio contents.The insulin,5-hydroxytryptamine(5-HT),and dopamine(DA)levels in the hippocampus were detected using an enzyme-linked immunosorbent assay.Immunofluorescence was used to detect the average fluorescence intensity of CD

关 键 词：糖尿病 抑郁症 左归降糖解郁方 小胶质细胞 糖代谢 白细胞单免疫球蛋白样受体3/葡萄糖转运体1信号通路 海马神经元 突触损伤 大鼠 

分 类 号：R285.5[医药卫生—中药学]