Studies On The Dynamic Protein Expression Profiles In Whole Brain Of Mice Subjeted To Chronic Mild Stress And Antidepressant-Anxiolytic Effects Of Wulingjun Powder

Depressive disorders are devastating metal illness that can lead to deterioration in the social and occupational functioning of affected individuals. Better understanding the mechanism of depressive disorder will facilitate the research and development of new drug. Adult BALB/c mice were exposed to chronic mild stress (CMS) for different periods and Differential2D gel electrophoresis (DIGE) approach was employed to the brain tissue to explore the molecular disease signatures.Wulingjun Capsule was approved as Class Ⅰ new drug by the Food and Drug Administration of China for the treatment of neurosis, insomnia, and morbid forgetfulness in adult. Systemic and scientific studies on the anxiolytic and anti-depressive effects of Wulingun Powder effects have not been reported. In this study, we aimed to extensively investigate the anxiolytic and antidepressive effects of Wulingjun Powder in behavioral mouse models. Meanwhile, the mechanism of Wulingun Powder was further investigated by cDNA microarray methods.Section I:Dynamic proteomic analysis of protein expression profiles in whole brain of Balb/c mice subjected to unpredictable chronic mild stress1. Adult BALB/c mice were exposed to the CMS for different periods and Differential2D gel electrophoresis (DIGE) approach was employed to the brain tissue to explore the molecular disease signatures.2. Sustained elevation of corticosterone level was observed Behavioral results indicated the depressive alterations of the mice exposing to CMS.3. The altered proteins identified by proteomics showed that abnormal energy mobilization under stress condition was accompanied by overproduction of reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress. Cytoskeleton protein and anti-oxidant enzymes were also changed by CMS treatment.4. These results indicated that the insufficiency of ATP synthesis, overwhelming ROS production and ER stress subsequently contributed to the cytoskeletal damage and inhibition to expression of some anti-oxidant proteins, which might ultimately bring functional neuron to apoptosis or death. Proteins whose expression is affected may provide tools for potential treatment strategies.Section II:Anxiolytic effects of Wulingjun Powder in behavioral models of anxiety1. Anxiolytic effect of Wulingjun Powder was assessed in the elevated plus-maze, the staircase and convulsion test. Male ICR mice were p.o.treated with Wulingjun Powder (0.25,0.50,1.00g kg-1bw.).2. Wulingjun Powder treatment elevated the%entries into open arms and%time on open arms compared with the control group In the elevated plus-maze test. Number of rearings in the staircase test was significantly decreased by Wulingjun Powder treatment. Wulingjun Powder (0.50g kg-1bw.) significantly elevated the latency of convulsion and depressed the seizure intensity induced by pentylenetetrazole (PTZ)(p<0.01). High dosage of Wulingjun Powder significantly lengthened the latency of seize inducd by picrotoxin (PTX).3. The current work demonstrates that treatment with Wulingun Powder can improve the anxiety symptom in male ICR mice.Section Ⅲ:Antidepressant effects of Wulingjun Powder in mice behavioral model1. Anti-depressant effect of Wulingjun Powder is assessed in the experimental animal model of depression, where male BALB/c mice were subjected to a CMS procedure involving9kinds of stressor for consecutive10weeks. Sucrose intake measurement, forced swimming test, Morris water maze, locomotor activities were employed. Additionally, the number of neuron and astrocyes was analyzed by immunohistochemistry method.2. Exposure to CMS procedure significantly reduced the sucrose intake, elevated the immobility time in the forced swimming test and escape latency in Morris water maze test. Meanwhile, the number of neron and astrocyte declined after CMS exposure. Wulingjun Powder treatment can reverse the changes mentioned aboved.3. The present results suggest that Wulingjun Powder exerts the antidepressant effects in mice. The machansim of Wulingjun Powder need futher investigation.Section Ⅳ:Anti-depressant mechanisms of Wulingjun powder revealed by gene microarray analysis1. An experimental depression model was established by exposing the mouse to aCMS procedure. The total RNA was extracted reverse-transcripted and hybrided to the mouse cDNA microarray. The difference of expression profiles were analyzed by the Image2.1software.2.130genes were significantly altered in stress group compared with the control groups. Among them,116genes were up-regulated and14genes were down-regulated. Meanwhile,85genes were significantly changed in the Wulingjun powder treated group with34genes up-regulated and51genes down-regulated compared with the model groups. For the Fluoxetine-treated group,133genes were significantly changed with35genes up-regulated and98genes down-regulated compared with the model groups.3. Multiple genes were affected by the stress exposure. Altered changes of some genes were normalized by Wulingjun powder and Fluoxetine treatment. In general, the mechanisms of Wulingjun powder and Fluoxetine are similar, but also with minor difference.