左归降糖解郁方对糖尿病并发抑郁症大鼠海马神经元GR、NR2A及NR2B的影响

The Effects of Zuogui Jiangtang Jieyu Formulation on hippocampus neuron GR,NR2A and NR2B duplicating diabetes mellitus complicated depression cell model

目的研究左归降糖解郁方(ZGJTJYF)对模拟糖尿病并发抑郁症(diabetes mellitus with depression,DD)环境下大鼠胎鼠海马神经元糖皮质激素受体(Glucocorticoid Receptor,GR)、N-甲基-D-天冬氨酸受体(N-methyl-D-aspartate,NR)亚基NR2A和NR2B的影响。方法分离、纯化、培养并鉴定胎鼠海马神经元(neuron,NE),将培养的海马神经元随机分为5组:空白组、空白血清组、模型组、阳性药物(二甲双胍+氟西汀)含药血清组和ZGJTJYF含药血清组。模型组、阳性药物组、ZGJTJYF组分别给予葡萄糖(150 mmol·L^(-1))和皮质酮(200μmol·L^(-1))用以构建模拟糖尿病并发抑郁症(DD)环境下海马神经元体外模型,阳性药物组和ZGJTJYF组分别给予10%相应含药血清,另空白血清组加入10%空白血清,干预18 h。采用高内涵细胞成像分析技术(High Content Analysis,HCA)检测GR、NR2A和NR2B蛋白的表达。结果与空白血清组相比,模型组GR蛋白表达降低,NR2A、NR2B蛋白表达升高;与模型组相比,阳性药物组和ZGJTJYF组GR表达上升,NR2A、NR2B蛋白的表达明显降低。结论左归降糖解郁方对模拟DD环境下海马神经元GR、NR2A及NR2B蛋白的调控作用,可能是抗DD大鼠神经元损伤的保护机制之一。

Objective To research the effects of Zuogui Jiangtang Jieyu Formulation（ ZGJTJYF） on Glucocorticoid Receptor（ GR） and N-methyl-D-aspartate Receptor（ NR） subunits NR2A,NR2B in fetal rat hippocampus neuron duplicating diabetes mellitus complicated depression（ DD）. Methods Separate,purify,culture and identify the neurons（ NE）,then divide them into five groups randomly： the blank group,blank serum group,model,melbine fluoxetine serum and ZGJTJF serum group. Exccept for the blank and blank serum groups,the other there groups apply for glucose（ 150 mmol·L^-1） and corticosterone（ 200μmol·L^（-1）） to build the model of diabetes mellitus complicated depression,offer 10% serum including medicine to melbine fluoxetine group and ZGJTJYF group,and offer blank serum group 10% serum,for 10 h. High content analysis tests the value of GR,NR2A and NR2B. Results Compared with blank serum group,the content of GR significantly cut down,the NR2A and NR2B raised in model group. Compared with model group,the content of GR increased,the NR2A and NR2B decreased in ZGJTJYF group. Conclusion ZGJTJYF could regulate the expression of GR,NR2A and NR2B in DD fetal rat hippocampus neuron,and the effect of ZGJTJYF may be one of the protective mechanisms of anti-apoptotic for DD hippocampus neuron.