锁阳总黄酮治疗围绝经期综合征的分子机制研究

Perimenopausal (Climacteric)Syndrome is a kind of clinical common diseases.Modern medicine is more used hormone replacement therapy to treat Perimenopausal Syndrome,Chinese medicine emphasize the tonifying kidney yang and filling essence and blood as the main therapeutic.Cynomorium is the product for tonifying kidney yang and benefiting essence and blood.Cynomorium is commonly used for curing Perimenopausal Syndrome,and according to report it has good resemble estrogen effect.The prophase research which we did shows that there is exactly curative effect on Cynomorium for Perimenopause (Climacteric) model.Total Flavonoids of Cynomorium is the main active site. This study aims to extraction and purification the Cynomorium to get the total Flavonoids of Cynomorium.Select not fully castrated rats to copy Perimenopause model and not fully castrated combined with chronic unforeseeable stress methods to copy the rats on Perimenopausal depression model as the research object.To observe the effect of total Flavonoids of Cynomorium on level of estradiol(E2),testosterone(T) in blood serum,ER and AR of relevant tissue, the influence of ERmRNA expression and the expression of Bcl-2、Bax on the hypothalamus and the pituitary,determine the level ofβ-EP in the plasma and gonadotropin releasing hormone（GnRH） and follicle stimulating hormone (FSH) and luteinizing hormone(LH) and interleukin-2(IL-2) and bone gla protein (BGP) in blood serum ,the influence of norepinephrine(NE) and 5-hydroxytryptamine (5-HT) and dihydroxyphenyl ethylamine(DA)，the tissue form and ultrastructure on the hypothalamus and pituitary and ovarian and uterus etc.In order to explore the related characteristics about the imbalance of the level of male and female hormone and corresponding receptor with related index,and reveal the molecular mechanism on the total Flavonoids of Cynomorium for treating Perimenopausal Syndrome.

围绝经期（更年期）综合征是临床常见病症之一，现代医学多采用激素替代疗法，中医以补肾阳、填精血为主要治则，锁阳为补肾阳、益精血之品，常用于围绝经期综合征的治疗，有报道其有好的拟雌激素样作用；我们前期研究表明，锁阳对围绝经期（更年期）模型疗效确切，锁阳总黄酮为其活性部位。本研究旨在从锁阳中提取纯化得到锁阳总黄酮，选择不完全去势造大鼠围绝经期模型及不完全去势结合慢性不可预见应激造大鼠围绝经期抑郁模型为研究对象，观察锁阳总黄酮对血清E2、T水平及相关组织ER、AR水平的影响，对下丘脑、垂体ERmRNA表达及Bcl-2、Bax表达的影响，对血浆β-EP及血清GnRH、LH、FSH、IL-2、BGP水平的影响，对脑中NE、5-HT、DA水平及下丘脑、垂体、卵巢、子宫等组织的形态及超微结构的影响。探讨雌雄激素及相应受体水平的失调与相关指标的关联特点，揭示锁阳总黄酮治疗围绝经期综合征的分子机制。

围绝经期综合征（PMS）是临床常见病。中医以补肾阳、填精血为主要治则，锁阳补肾阳、益精血，常用于PMS的治疗。前期研究发现锁阳对PMS模型疗效确切，锁阳总黄酮为其活性部位，但机制不清。. 受国家基金支持，从锁阳中提取纯化得到71％的锁阳总黄酮，继而开展药效研究。不完全去势造大鼠PMS模型成功。锁阳总黄酮可显著改善模型大鼠的行为学、生化、病理及基因表达相关指标，明显或显著升高模型大鼠降低的糖水消耗量、水平-垂直积分、站立次数、子宫（胸腺、脾脏）指数、显著缩短模型延长的强迫游泳不动时间，明显或显著升高模型降低的血清E2、IL-2、BGP水平，显著降低模型升高的血清FSH、LH水平；明显或显著升高模型降低的下丘脑、垂体中AR的表达，下丘脑、垂体、子宫、卵巢中ER的表达和下丘脑ERmRNA表达；显著升高模型降低的下丘脑、垂体Bcl-2表达，降低模型升高的Bax表达；显著改善模型导致的子宫、胸腺、脾脏、下丘脑的病理变化。. 不完全去势结合慢性不可预见刺激造大鼠围绝经期合并抑郁模型成功。锁阳总黄酮可显著改善模型大鼠的行为学、生化、病理及基因表达相关指标，明显或显著升高模型降低的大鼠水平运动距离、站立次数、糖水消耗量、子宫（胸腺、脾脏）指数，显著缩短模型延长的强迫游泳不动时间；明显或显著升高模型降低的血清E2、T、BGP、IL-2、血浆β-EP水平，脑匀浆NE、5-HT、DA水平；明显或显著降低模型升高的FSH、LH、GnRH水平；显著升高模型降低的下丘脑ERmRNA表达，下丘脑、垂体AR、Bcl-2表达、下丘脑、垂体、子宫、卵巢ER表达和下丘脑ERmRNA的表达； 显著升高模型降低的下丘脑、垂体Bcl-2表达、显著降低模型升高的Bax表达；显著改善模型导致的下丘脑、垂体、子宫、卵巢、胸腺、脾脏的病理变化。. 本研究揭示了锁阳总黄酮治疗PMS的分子机制。发表核心期刊论文5篇、SCI收录论文4篇，EI收录1篇，出版学术专著2部，培养研究生3名，获得2项发明专利，新申请1项发明专利。研制了治疗PMS的归锁胶囊医院制剂，为PMS的治疗提供一种新思路。圆满完成原定目标任务。