左归降糖解郁方对模拟DD环境致胎鼠海马神经元凋亡保护作用的研究

Zuoguijiangtangjieyu Formulation Prevents Apoptosis of Hippocampal Neurons Induced by Mimetic DD Condition

目的研究左归降糖解郁方对模拟糖尿病并发抑郁症(diabetes mellitus with depression,DD)环境下海马神经元凋亡的保护作用。方法从胎鼠海马组织中分离、纯化和培养海马神经元,采用神经元特异性烯醇化酶(NSE)鉴定海马神经元。采用高糖(150m M)联合皮质酮(200μM)诱导复制模拟DD环境海马神经元损伤模型。实验分为空白对照组、空白血清组、高糖联合皮质酮组、二甲双胍0.18g/kg+氟西汀1.8mg/kg(M+F)组、左归降糖解郁方32.82g/kg(ZGJTJYF)组,造模及药物干预18h,采用MTT法,流式细胞术,高内涵成像分析系统检测各组海马神经元损伤及干预后活力及凋亡的变化情况。结果与空白对照组比较,高糖联合皮质酮组海马神经元胞体破裂,神经网络断裂,细胞活力减小,凋亡显著(P<0.05);ZGJTJYF组海马神经元胞体圆润,神经网络丰富,细胞活力较强,凋亡率小,与模型组比较,差异有统计学意义(P<0.05)。空白血清组细胞状态和细胞活力与正常对照组比较,无明显差异,与模型组比较,差异有统计学意义(P<0.05)。M+F组细胞状态和细胞活力与模型组比较有明显差异(P<0.05),与ZGJTJYF组比较不具有统计学意义。结论左归降糖解郁方能增强模拟DD环境下海马神经元的活力,减少其凋亡,对模拟DD环境下海马神经元具有保护作用。

Objective To investigate the Zuoguijiangtangjieyu Formulation（ZGJTJYF） on apoptosis of hippocampal neurons in- duced by mimetic diabetes mellitus with depression （DD） condition. Methods Hippocampal neurons isolated from fetal rat hippo- campus, and then we purified and cultured them. Neuron -specific enolase （NSE） was usd identified hippocampal neurons. High concentrations of glucose （150raM） combined with corticosterone （200p, M） induced damage of hippocampal neurons in mi- metic DD condition. Hippocampal neurons were divided into control group , blank control serum group, model group , Metformin ＋ Fluoxetine group （M ＋ F） and ZGJTJYF group. Hippocampal neurons were induced injury and drug intervention for 18 hours. MTT assay, flow cytometry, high - content image analysis system were used to detect changes in hippocampal neurons in each group after injury and intervention vitality and apoptosis. Results Compared with the control group, hippocampal neurons were rupture, neural network fault, neurons viability decreased and apoptosis significantly in model group （ P 〈 0. 05 ）. Compared with the model , hippocampal neurons in ZGJTJYF group were mellow, had rich neural networks and small apoptosis rate, and cell via- bility is high, the difference was statistically significant （ P 〈 0.05 ）. Cell morphology and cell viability of Hippocampal neurons in blank control serum group had no significant difference compared with the control group. There difference was statistically sig- nificant when it compared with the model group （ P 〈 O. 05 ）. Conclusion ZGJTJYF can enhance the vitality of damaged hipp- ocampal neurons in mimetic DD condition , reduce apoptosis, and has a protective effect on hioooeamoal neurons.