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
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摘要
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(50 μmol/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.
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(50 μmol/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.
引文
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[13]王宇红,杨蕙,尹玲桃,等.左归降糖解郁方对糖尿病并发抑郁症大鼠血糖血脂的影响[J].中草药,2014,45(11):1602-1606.
[14]凌佳,杨琴,刘检,等.左归降糖解郁方对模拟糖尿病并发抑郁症环境下模型大鼠海马神经元凋亡相关蛋白的影响[J].中国中医药信息杂志,2017,24(10):35-39.
[15]ZHANG J,YANG Y,LI H,et al.Amplitude/frequency of spontaneous m EPSC correlates to the degree of long-term depression in the CA1region of the hippocampal slice[J].Brain Res,2005,1050(1/2):110-117.
[16]LONSKAYA I,PARTRIDGE J,LALCHANDANI R R,et al.Soluble ICAM-5,a product of activity dependent proteolysis,increases m EPSC frequency and dendritic expression of Glu A1[J].PLo S One,2013.8(7):e69136.
[2]HUNTER J C,DEVELLIS B M,JORDAN J M,et al.The association of depression and diabetes across methods,measures,and study contexts[J].Clin Diabetes Endocrinol,2018,4:1.
[3]MOURI A,UKAI M,UCHIDA M,et al.Juvenile social defeat stress exposure persistently impairs social behaviors and neurogenesis[J].Neuropharmacology,2018,133:23-37.
[4]HO N,SOMMERS M S,LUCKI I.Effects of diabetes on hippocampal neurogenesis:links to cognition and depression[J].Neurosci Biobehav Rev,2013,37(8):1346-1362.
[5]WANG Y H,YIN LT,YANG H,et al.Hypoglycemic and anti-depressant effects of Zuogui Jiangtang Jieyu formulation in a model of unpredictable chronic mild stress in rats with diabetes mellitus[J].Exp Ther Med,2014,8(1):281-285.
[6]刘检,王宇红,徐雅岚,等.改良的胎鼠海马神经元原代培养及模拟糖尿病并发抑郁环境对其的损伤作用[J].神经解剖学杂志,2016,32(4):459-465.
[7]孟盼,赵洪庆,杜青,等.左归降糖解郁方对糖尿病并发抑郁症大鼠海马神经元凋亡的影响[J].中国中医药信息杂志,2016,23(9):78-81.
[8]李珊,胡喆,周燕,等.趋化因子MCP-1对大鼠海马区NMDA受体介导的兴奋性突触后电流的影响[J].中国药理学通报,2016,32(7):950-955.
[9]LISMAN J.Glutamatergic synapses are structurally and biochemically complex because of multiple plasticity processes:long-term potentiation,long-term depression,short-term potentiation and scaling[J].Philos Trans R Soc Lond B Biol Sci,2017,372(1715):20160260.
[10]ASALGOO S,TAT M,SAHRAEI H,et al.The psychoactive agent crocin can regulate hypothalamic-pituitary-adrenal axis activity[J].Front Neurosci,2017,11:668.
[11]WANG Y,YANG H,LI W,et al.Zuogui Jiangtang Jieyu formulation prevents hyperglycaemia and depressive-like behaviour in rats by reducing the glucocorticoid level in plasma and hippocampus[J].Evid Based Complement Alternat Med,2015,2015:158361.
[12]杨蕙,王宇红,杜青,等.左归降糖解郁方对糖尿病并发抑郁症大鼠海马星形胶质细胞神经营养的影响[J].中国中医药信息杂志,2017,24(9):43-47.
[13]王宇红,杨蕙,尹玲桃,等.左归降糖解郁方对糖尿病并发抑郁症大鼠血糖血脂的影响[J].中草药,2014,45(11):1602-1606.
[14]凌佳,杨琴,刘检,等.左归降糖解郁方对模拟糖尿病并发抑郁症环境下模型大鼠海马神经元凋亡相关蛋白的影响[J].中国中医药信息杂志,2017,24(10):35-39.
[15]ZHANG J,YANG Y,LI H,et al.Amplitude/frequency of spontaneous m EPSC correlates to the degree of long-term depression in the CA1region of the hippocampal slice[J].Brain Res,2005,1050(1/2):110-117.
[16]LONSKAYA I,PARTRIDGE J,LALCHANDANI R R,et al.Soluble ICAM-5,a product of activity dependent proteolysis,increases m EPSC frequency and dendritic expression of Glu A1[J].PLo S One,2013.8(7):e69136.