雌激素对中脑多巴胺能神经元的保护作用及其机制分析-protective effect of estrogen on dopaminergic neurons in midbrain and its mechanism analysis.docxVIP

第三军医大学硕士学位论文第三军医大学硕士学位论文33effects of estrogen on dopaminergic neurons. Adult（3-4 month）female C57B/L6 mice wererandomized into four groups : control group (intact mice), sham group , vehicle group（ovariectomized mice with sesame oil ip injections）and 17β-estradiol group（ovariectomized mice with 17β-estradiol ip injections ） . The neuroprotection effects of estrogen ondopaminergic neurons were investigated by measurement of DAT using RT-PCR and immunohistochemistry staining of tyrosine hydroxylase(TH) ， changes of astrocyte andinterneurons were detected by immunohistochemistry staining of glial fibrillary acidic protein(GFAP) and Calretinin. The molecules involed the effect of estrogen such as ERK and AKT were detected with Western blot .The results were as follows:Immunohistochemistry demonstrated that both of the estrogen receptor subtypes were expressed in the substantia nigra, ERβ appears to be the predominant subtype in this area.The gene expression level of ERβ significantly decreased in ovariectomized mice comapared with control group, treatment of ovariectomized mice with estrogen can prevented the reduction. The gene expression level of ERα shows no change in all groups.The number of TH-IR neurons and TH-calretinin double staining neurons significantly decreased in ovariectomized mice comapared with control group mice, treatment of ovariectomized mice with estrogen can prevent the reduction of the positive neurons.Comapared with control group,the gene expression level of DAT、SYN、BDNFsignificantly decreased in ovariectomized mice, treatment of ovariectomized mice with estrogen can prevented the reduction.Comapared with control group, the number of astrocyte did not changed in four groups and no morphological changes were observed.The protein expression level of ERK and phosphorylated ERK was evaluated by Western blot ,but no significant changes were observed in four groups.Comapared with control group, estrogen treatment can increase