左归降糖解郁方对糖尿病并发抑郁症大鼠海马神经元微小兴奋性突触后电流的影响

Effects of Zuogui Jiangtang Jieyu Prescription on Miniature Excitatory Postsynaptic Current of Hippocampal Neurons in Diabetes Mellitus Rats with Depression States

目的观察左归降糖解郁方对糖尿病并发抑郁症大鼠海马神经元微小兴奋性突触后电流(mEPSC)频率及幅度的影响,探讨其对海马神经元保护作用的可能机制。方法选用17~19 d胚胎大鼠离体培养海马神经元,神经元特异性烯醇化酶(NSE)免疫细胞化学染色进行细胞鉴定。细胞随机分为空白对照组、抑郁组、糖尿病组、糖尿病并发抑郁症组、空白血清组、左归降糖解郁方组和阳性药组,培养10 d后加入皮质酮(50μmol/L)和/或高糖(15 mmol/L)建立细胞模型,给予药物血清干预18 h,采用全细胞膜片钳记录神经元mEPSC,比较各组神经元mEPSC频率和电流幅度。结果经NSE免疫细胞化学鉴定,培养7 d后神经元纯度达95%以上。与空白对照组比较,抑郁组、糖尿病组和糖尿病并发抑郁症组大鼠海马神经元mEPSC的频率和幅度均显著上升(P<0.01);与糖尿病并发抑郁症组比较,左归降糖解郁方组和阳性药组大鼠海马神经元mEPSC的频率和幅度均显著下降(P<0.01)。结论左归降糖解郁方可降低糖尿病并发抑郁症大鼠海马神经元mEPSC的频率和幅度,对海马神经元有保护作用。

Objective To investigate the effects of Zuogui Jiangtang Jieyu Prescription on the frequency and amplitude of miniature excitatory postsynaptic current(mEPSC)of hippocampal neurons cultured in diabetes mellitus with depression(DD)states;To discuss the possible mechanism for the protection of hippocampal neurons.Methods Hippocampal neurons of 17–19 d embryonic rats were cultured.Immunocytochemical staining for neuron specific enolase(NSE)was performed to identify hippocampal neurons.The cells were randomly divided into blank control group,depression group,diabetes mellitus group,DD group,blank serum group,Zuogui Jiangtang Jieyu Prescription group and positive medicine group.After culturing for 10 days,cortisone(50μmol/L)and/or D-glucose(15 mmol/L)were added to establish the cell model.The medicine serum intervened for 18 h.Whole-cell patch-clamp recording was performed to record the mEPSC of neurons.The difference of frequency and amplitude of mEPSC was compared between each groups.Results The immunocytochemical staining for NSE showed that the purity of neurons cultivated for 7 d was over 95%.Compared with the blank control group,the frequency and amplitude of mEPSC in hippocampal neurons of depression group,diabetes mellitus group and DD group were significantly increased(P<0.01).Compared with DD group,the frequency and amplitude of mEPSC in hippocampal neurons of Zuogui Jiangtang Jieyu Prescription group and the positive medicine group were significantly decreased(P<0.01).Conclusion Zuogui Jiangtang Jieyu Prescription can reduce the mEPSC frequency and amplitude of hippocampal neurons in DD states,and have a protective effect on hippocampal neurons.