左归降糖解郁方对模拟糖尿病并发抑郁症环境下海马神经元的影响

目的观察左归降糖解郁方对模拟糖尿病并发抑郁症(DD)环境下海马神经元的影响,探讨其可能的作用机制。方法海马神经元原代细胞取自受孕18 d SD大鼠胎鼠,葡萄糖联合皮质酮干预构建DD海马神经元细胞模型。将培养的海马神经元随...展开更多 目的观察左归降糖解郁方对模拟糖尿病并发抑郁症(DD)环境下海马神经元的影响,探讨其可能的作用机制。方法海马神经元原代细胞取自受孕18 d SD大鼠胎鼠,葡萄糖联合皮质酮干预构建DD海马神经元细胞模型。将培养的海马神经元随机分为正常组、空白血清组、模型组、阳性药(二甲双胍+氟西汀)药物血清组、中药(左归降糖解郁方)药物血清组和阻断剂组。正常组和模型组给予等量培养液,阻断剂组给予促肾上腺皮质激素释放激素1型受体(CRHR1)阻断剂安塔拉明200μmol/L,其余组给予体积分数为10%的药物血清或空白血清,造模干预18 h后,尼氏染色检测海马神经元损伤情况,高内涵细胞成像分析(HCA)技术检测CRHR1、突触可塑性相关蛋白切丝蛋白(Cofilin)和肌动蛋白(Actin)蛋白的表达,实时荧光定量PCR检测CRHR1、Cofilin和Actin mRNA的表达。结果与正常组及空白血清组比较,模型组海马神经元神经网络、树突棘断裂或消失,尼氏小体数量显著减少,CRHR1表达明显上调(P<0.01),Cofilin和Actin表达明显下调(P<0.05);与模型组比较,阳性药药物血清组、中药药物血清组和阻断剂组可一定程度上逆转上述变化(P<0.05,P<0.01)。结论左归降糖解郁方对模拟DD环境下海马神经元具有保护作用,其机制与调控CRHR1、Cofilin、Actin的表达有关。收起 Objective To explore the effects ofZuogui Jiangtang Jieyu Prescription on hippocampal neurons in simulated diabetes mellitus with depression (DD);To discuss its possible mechanism.Methods Hippocampal neurons from fetal rats of 18 d pregnant rats were primitively cultiva...MORE Objective To explore the effects ofZuogui Jiangtang Jieyu Prescription on hippocampal neurons in simulated diabetes mellitus with depression (DD);To discuss its possible mechanism.Methods Hippocampal neurons from fetal rats of 18 d pregnant rats were primitively cultivated,and the glucose combined with corticosterone was used to establish DD hippocampal neuronal cell model.Cultured hippocampal neurons were randomly divided into normal group,blank serum group,model group,positive drug (metformin plus fluoxetine) serum group,TCM (Zuogui Jiangtang Jieyu Prescription)serum group and blocker group.The normal group and the model group were given equal amount of culture,the blocker group was given 200μmol/L Antalarmin,and the remaining groups were given 10% volume of drug serum or blank serum.After modeling intervention for 18 h, Nissl staining was used to detect hippocampal neurons injury.High content analysis (HCA)was used to detect the expressions of CRHR1,Cofilin and Actin protein,and real-time PCR was used to detect expressions of CRHR1,Cofilin and Actin mRNA.Results Compared with the normal group and blank serum group,neural network and dentritic spines ruptured or disappeared and the quantity of Nissl bodies decreased significantly,and expression of CRHR1 was significantly up-regulated (P<0.01),while the expressions of Cofilin and Actin were significantly down-regulated (P<0.05).Compared with the model group,positive drug serum group,TCM serum group and blocker group could partially reverse the above process (P<0.05,P<0.01).Conclusion Zuogui Jiangtang Jieyu Prescription has protective effects on hippocampal neurons in simulated DD environmen,and its mechanism is related to the regulation expressions of CRHR1,Cofilin and Actin.FEWER

作者 金狮 黄会珍 凌佳 刘检 柳卓 罗薇絮 吴梦瑶 王宇红

Effects of Zuogui Jiangtang Jieyu Prescription on Hippocampal Neurons in Simulated Diabetes Mellitus with Depression

JIN Shi;HUANG Huizhen;LING Jia;LILT Jian;LIU Zhuo;LUO Weixu;WU Mengyao;WANG Yuhong(Hunan Key Laboratory of Power and Innovative Drugs State Key Laboratory of Ministry Training Bases, Hunan University of Chinese Medicine,Changsha 410208,China;Central Laboratory,First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)