Study On Apoptosis Of Hippocampus Neuron And The Interventional Mechanism Of Chinese Medicinal In Diabetes Mellitus With Depression Rats

Objective:1.To investigate the molecular mechanism of hippocampal apoptosis in diabetes mellitus with depression rats,preliminary explore the pathogenesis of diabetes mellitus with depression.2. To explore the effect of Zuo Gui Jiang Tang Jie Yu prescription., what was a traditional Chinese medicine compound prescription on the animal model of diabetes mellitus with depression, and further to analysis the effect of its anti-apoptotic hippocampal. This study will lay the foundations for diabetes mellitus with depression experimental study.Method:1.The study of hippocampal apoptosis in diabetes mellitus with depression rats:SD rats were received a high fat diet for14days before the intraperitoneal injection of streptozotocin (38mg/kg). The rats were randomly divided into2groups according to their behavior, the diabetes group and diabetes mellitus with depression group; the normal rats were also randomly divided into2groups according to their behavior, normal group and depression group. Assayed the body weight of rats in each group after28days chronic stress.Blood glucose and Glycated hemoglobin were measured by biochemical techniques. Open-field test and Morris water maze were used to evaluate the behaviors of rats. HE staining was used to observe the hippocampus pathological changes of rats in each group. Western-blotting was used to detect the expression of p-NR2A、NR2A、 p-NR2B、NR2B、p-JNK (Thr183)、JNK1/2、Bax, Bcl-2、Caspase8and Caspase3proteins on hippocampal.RT-PCR was used to detect the expression of NR2A、NR2B、JNK、Bax, Bcl-2、 Caspase8genes on hippocampal. 2.The effect of Zuo Gui Jiang Tang Jie Yu prescription. DD was established by the combination of high-fat diet,injection of Streptozotocin(STZ, mg-kg-1), and chronic stress.The model rats were randomly divided into five groups,the diabetes mellitus with depression group, metformin combined fluoxetine group, Zuo Gui Jiang Tang Jie Yu prescription high, medium and low dose group. After4weeks, the the above indicators of established evaluation system were used to measured for rats in each group.Results:1. Compared with the normal group, the body weight of diabetes mellitus with depression rats were significantly reduced (P＜0.01), the locomotor activity of model group were significantly reduced (P<0.01), the escape latency was significantly longer in the Morris water maze test, as well as the space exploration time was significantly shortened(P <0.05).Blood glucose and Glycated hemoglobin were significantly higher (P＜0.01), HE staining observed the hippocampus existed the significant pathological changes. Compared with the normal group, The p-NR2A、 NR2A、p-NR2B、NR2B、p-JNK (Thr183)、JNK1/2、Bax、Caspase8. Caspase3expression of hippocampal were significantly increased(P<0.01) and Bcl-2was reduced(P<0.05) in model group through Western-blotting. The NR2A、NR2B、JNK、Bax、Caspase8mRNA expression of hippocampal were significantly increased(P<0.01) and Bcl-2mRNA was reduced (P＜0.01) in model group through RT-PCR.2. Compared with the normal group, the above indicators of model group were significantly abnormal(P＜0.05). Compared with the model group,the body weight of high dose group rats with Zuo Gui Jiang Tang Jie Yu prescription were significantly increased(P<0.05),the locomotor activity were significantly increased(P<0.01), the spatial leaning and memory ability to be a certain degree of recovery(P<0.05), the blood glucose and Glycated hemoglobin were significantly stable(P<0.05),and the hippocampal pathological changes was mitigated.The p-NR2A、NR2A、 p-NR2B、NR2B、p-JNK1(Thr183)×JNK1/2Bax、Caspase8and Caspase3expression were decreased (P<0.05),as well as Bcl-2was increased(P<0.05). The NR2A、NR2B、JNK、Bax、Caspase8mRNA expression of hippocampal were significantly reduced (P<0.05) and Bcl-2mRNA was increased(P<0.05) through RT-PCR.The middle dose and low dose of Zuo Gui Jiang Tang Jie Yu prescription had a weak effect than the high dose group.Conclusion:1. Hippocampus was serious injury, and there were elevation in expression levels of p-NR2A、NR2A、p-NR2B、NR2B protein in the hippocampus of diabetes mellitus with depression rat,which suggests that NR might be the key targets of hippocampus neuronal apoptosis lesion of diabetes mellitus with depression.2.The mainly possible mechanism of DD is the over-expression of NR,which could activated JNK and JNK downstream apoptotic pathway,Mediated hippocampus apoptosis and futher leads to depression.3. The general symptoms could be improved by ZJJF,which might be correlated to decrease NR receptor,Bax and caspase8proteins over expression, inhibition of neuronal apoptosis for protecting the hippocampal.