| Objectives:The study was conducted to explore the effects of Baishile capsule on ethology, the morphology and cell proliferation of hippocampus, as well as Cx32, Cx43and SYP expressions of hippocampus in Chronic stress depression rats, and to explore the mechanisms of Baishile capsule remodeling hippocampus nerve of the chronic stress depression rats, which providing experimental evidences for Baishile capsule in the treatment of clinical depression drug.Methods:Seventy-two SD rats were assigned randomly into four groups (18rats/group, half male and half female), normal control group, model group, fluoxetine group (fluoxetine hydrochloride group), and Baishile group (Baishile capsule group). Rats of model group, fluoxetine group and Baishile group were oral administrated with distilled water, fluoxetine hydrochloride soup (1.8mg·kg-1·d-1) and Baishile capsule soup (contented with1.33g·kg-1·d-1), respectively, for21days. Six rats were selected randomly from each group to intraperitoneal inject BrdU (100mg·kg-1·d-1) for proliferation cell markers. All rats were weight, horizontal and vertical activities and sugar consumptions were tested before and after molding, and after oral administration. Six cerebral tissues were selected randomly to make paraffin sections from each group, and then morphology changes of hippocampal cell and functional changes of hippocampal nerve were observed using HE staining and Nissl staining, respectively, Cx32, Cx43and SYP expressions were tested using immunohistochemical staning. And6cerebral tissues were selected to make frozen sections for hippocampal cell proliferation determination via BrdU immunohistochemistry staining. Hippocampus were separated from6cerebral tissues and protein were extracted for Cx32, Cx43and SYP protein expressions analyses using Western blot.Results:(1)Compared with the normal group, the body weight, the scores of rearing and crossing and sweet water consumption of rats in model group were all decreased (P<0.01or P<0.05). The body weights of rats in Baishile group were increased (P<0.05), the horizontal (P<0.01) and vertical (P<0.05) activities remarkably improved and sweet water consumption were obviously increased (P<0.01) than that of the model group.(2) From the results of HE staining, the abnormal changes of the neurons in hippocampal CA1and CA3areas of the model group rats were observed, which exhibiting shrinking cell, irregular nuclei, scattered cell arrangement and more neuron turned degeneration, karyopyknosis and deeper color, lager intercellular gap. However, in the Baishile group, neuron morphology structure of is close to that of the normal group with integrity cell structure, neatly arranged, big and round nuclei, clear nucleolus and only a little of nuclei degeneration.(3) The results of Nissl staining showed that some neuronal cell outlines of hippocampal CA1, CA3and DG areas in the model group rats were blur, fewer Nissl bodies, lighter color, larger intercellular while neronal cells arranged in uniform, outlines were clear, the number of Nissl bodies were rich, the cytoplastic was well-distributed and has deep color in Baishile group, which is closed to normal group.(4) The results of BrdU immunohistochemistry showed that there were a certain numbers of proliferating cells in hippocampal CA1, CA3and DG region of all groups and the numbers of DG district proliferating cells were significantly larger than that of other two regions. The numbers of proliferating cells in hippocampal CA1, CA3and DG region of the Baishile group increased (P<0.05) significantly compared with the model group and fluoxetine group.(5) The results of IHC and WB showed that gap junction proteins Cx32and Cx43expression levels as well as the average optical of synaptophysin SYP density and the ratio of its optical density to β-actin in the hippocampus of model group rats were significantly lower (P<0.01or P<0.05) than that of normal group; Cx32, Cx43and SYP protein expression levels in the hippocampal of Baishile group rats were significantly increased (P<0.01or P<0.05) compared with that of model group.Conclusions:(1) Baishile capsule can increase body weight, spontaneous the activity and the ability to explore the novel environment, as well as the reaction of the happy event of the chronic stress depression rats.(2) Baishile capsules have neuroprotective effects, which reducing chronic stress-induced atrophy and apoptosis of rat hippocampal neurons and recovering hippocampal nerve function.(3) Baishile Capsules promote hippocampal nerve regeneration of chronic stress induced depression rat, and its mechanism may be related to the activation of NPCs in the hippocampal DG region.(4) The up-regulations of Cx32, Cx43and SYP expressions may be an important mechanism of Baishile Capsules improving plastic change of hippocampal neurons in the chronic stress depression rats.(5) Baishile capsule enhanced hippocampus of Cx43expression may be partly resulting to promoting nerve regeneration. |
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