Establishment Of The Protective Effect Of Traditional Chinese Medicine In Patients With Depression Diabetes Cell Model

Objective:1. To select the appropriate intervent concentration of glucose combined with corticosterone, and establish a cell model of diabetes mellitus with depression, and establish a model evaluation system from the cell morphology and neurochemistry.2. To explore the therapeutic mechanism of Zuo Gui Jiang Tang Jie Yu prescription, what was a traditional Chinese medicine compound prescription on the cell model of diabetes mellitus with depression, and to further clarify the effect of its hypoglycemic and anti-depression. This research will reveal the foundations of diabetes mellitus with depression for experimental study.Method:1. To build a cell model of diabetes mellitus with depression.(1) To select the inductor of diabetes mellitus with depression. In vitro culture of primary hippocampal neurons7d, by the difference of inductor were randomly divided into nine groups. Through the survival rate of hippocampal neurons to choose the best intervent concentration of glucose combined with corticosterone.(2) To build a cell model of diabetes mellitus with depression. According to the best combination of (1), establish the DD cell model validation experiments. By the difference of interventional drugs were randomly divided into four groups: normal control group (no treatment); glucose group (75mM/L glucose intervention); corticosterone group (100μM/L corticosterone intervention); glucose combined corticosterone group (75mM/L glucose combined100μM/L corticosterone intervention). After drug intervention24h, morphological evaluation, identification of NSE nerve cells, MTT cell viability, TUNEL staining was measured apoptotic bodies, glucose oxidase-peroxidase (GOD-POD) method to detect glucose consumption hippocampal neurons value; ELISA assay neurotransmitter serotonin (5-HT) immunocytochemistry to detect the hippocampal neurons neurotrophic factor CREB and BDNF and key proteins NR2A, NR2B expression.2. The effect of Zuo Gui Jiang Tang Jie Yu prescription. The diabetes mellitus with depression hippocampal neurons were randomly divided into four groups: normal group, model group, model+(metformin+Prozac) drug-containing plasma group, model+Zuo Gui Jiang Tang Jie Yu prescription drug-containing plasma group. After drug intervention24h, the indicators of established evaluation system were used to measured for hippocampal neurons in each group.Results:1.(1) Compared with the normal group, cell survival rate decreased significantly in the corresponding time points and smooth of the seventh group (75mM/L glucose combined with100μM/L corticosterone).(2) Compared with the normal group, the hippocampal neuron viability of Glucose (75mM/L) combined with corticosterone (100μM/L) group were decreased significantly(P <0.01); the number of apoptotic bodies were significant increased; glucose consumption values were decreased significantly (P<0.01); the5-HT was decreased significantly(P<0.01); intracellular neurotrophic factor CREB and BDNF were significantly lower (P<0.05).2. Compared with the normal group, the indicators of model group were significantly abnormal. Compared with the model group, hippocampal neurons of Zuo Gui Jiang Tang Jie Yu prescription drug-containing plasma group in cell survival were increased significantly after24h (P<0.05); the number of apoptotic bodies were decreased significantly; glucose consumption values were increased significantly (P<0.01) after12h; the levels of5-HT, NE and DA were increased significantly (P<0.01); CREB and BDNF were significantly increased (P<0.05); the expression of NR2A of key proteins were decreased, NR2B expression were increased (P<0.01).Conclusion:1. Combining literature research and experimental results, the choice of75mM/L glucose combined100μM/L corticosterone as the best intervent concentration of glucose combined corticosterone.2. Combination the clinical features of diabetes mellitus with depression, GOD-POD method to detect glucose consumption values, neurotransmitters and neurotrophic factor can create an evaluation system of diabetes mellitus with depression animal models.3. The composite method of glucose (75mM/L) combined corticosterone (100μM/L) intervention in primary hippocampal neurons mature cells24hours can be successfully prepared animal models of diabetes mellitus with depression.4. Hippocampal neurons of diabetes mellitus with depression were serious injury-and theirs neurotransmitter secretion disorders, and the content of corticosterone abnormally elevated lead to hippocampal neurons’pathological changes.The down regulation of CREB and BDNF in the intracellular neurotrophic factor are likely to participate in the damage of hippocampus, and to promote the development of diabetes mellitus with depression.5. The Zuo Gui Jiang Tang Jie Yu prescription can increase the survival rate of hippocampal neurons, reducing the number of apoptotic bodies, regulate glucose consumption, improve its neurotransmitter secretion disorders, increased expression of neurotrophic factors, such as, Its therapeutic effect may be associated with the increased the key protein expression of NR2A and NR2B. The treatment of Zuo Gui Jiang Tang Jie Yu prescription for diabetes mellitus with depression to be worthy of further research and development.