针灸对围绝经期抑郁症单胺类神经递质影响的关联性研究

单胺类神经递质属于中枢神经递质,是一类生物学活性比较广泛的物质,单胺类神经递质含量的平衡失调与围绝经期抑郁症的发病密切相关。中医针灸治疗围绝经期抑郁症已取得很好的疗效,同时对单胺类神经递质具有调节作用,本文通过研究针灸、围绝经期抑郁症以及单胺类神经递质三者之间的关联性,进一步明确中医针灸治疗围绝经期抑郁症可能的机制之一,为临床治疗提供理论依据。     

安神方对大鼠抑郁模型行为学及单胺类神经递质的影响

目的探讨中药复方安神方抗抑郁作用的机制。方法以慢性中等强度应激刺激建立大鼠抑郁模型,用强迫游泳试验进行行为学评分,用高效液相电化学检测方法检测大鼠脑干内单胺类神经递质的含量,观察模型大鼠给药前后的变化。结果抑郁模型大鼠第7天与第28天水中强迫游泳不动时间分别为（133.4±24.0）s、（179.7±35.3）s,正常组则为（127.9±31.3）s、（131.8±29.3）s,模型组显著高于正常组（P〈0.05）;脑干内5-HT、NE含量显著降低,模型组为（0.093±0.031）ng/mg、（0.146±0.038）ng/mg,正常组为（0.160±0.023）ng/mg、（0.237±0.049）ng/mg（P〈0.05）;安神方高剂量组第7天与第28天水中强迫游泳不动时间分别为（121.5±27.8）s、（145.1±24.2）s,与模型组比较显著降低（P〈0.05）,大鼠脑干内5-HT、NE含量分别为（0.151±0.021）ng/mg、（0.208±0.057）ng/mg,较正常组显著提高（P〈0.05）。结论安神方具有抗抑郁作用,对中枢单胺类神经递质的调节作用是其疗效机制之一。     

载脂蛋白E基因缺陷小鼠雌激素与抑郁症及血脂相关性的初步研究

目的探讨载脂蛋白E基因缺陷（apo-/-）小鼠雌激素（E2）、血清总胆固醇水平（TC）和抑郁症之间的关系。方法 30只成年apo-/-雌性小鼠随机分为3组。A组：双侧卵巢切除（OVX）＋E2;B组：OVX＋芝麻油;C组：假手术组（Sham）。三个月后ELISA检测血清E2和TC水平,观察小鼠悬尾不动时间,分析他们之间的关系。结果小鼠悬尾不动时间和TC水平与E2呈明显的负相关,小鼠悬尾不动时间与TC有一定的正相关。结论雌激素能抑制抑郁症倾向和降低TC水平,三者关系中,雌激素起主导作用。     

雌激素对大鼠心脏雌激素受体α和β表达的影响

目的研究雌激素受体（ER）α和β在新生、成年和卵巢切除大鼠心脏的表达。方法采用半定量逆转录聚合酶链反应（RT-PCR）和Western blot方法,分析新生和成年大鼠心脏各部的ERα和βmRNA及ER蛋白质的表达,探讨不同剂量的17β雌二醇对卵巢切除大鼠心脏ERα和βmRNA、ER蛋白质的表达调控及对心房、心室重量和它们与体重比变化的影响。结果ERα mRNA在新生大鼠心脏各部表达较低,而成年大鼠心脏各部ERα mRNA的转录水平提高3～20倍,这种差别体现在ERα蛋白质的表达水平不同。反之新生大鼠心脏ERβ mRNA表达水平较高,而成年大鼠心脏ERβ mRNA的表达降低2～7倍。卵巢切除减少成年大鼠心房ERαmRNA和蛋白质的表达,减轻心房重量及心房／体重比;17肛雌二醇替代却能反转这些改变,同时随着血浆17β-雌二醇水平的增高,这种影响在一定程度上呈剂量依赖关系。卵巢切除和178一雌二醇替代并不改变心脏和心室／体重比,也不影响ERβ mRNA的表达。结论雌激素受体α和β在新生和成年大鼠心脏的表达存在差别,ERβ可能在大鼠心脏的发育过程中起重要作用,ERα足雌激素对成年大鼠心脏调控的优势受体;雌激素的主要靶器官是心房,且通过其受体的介导促进心房肌细胞的增生。     

运动对创伤后应激障碍大鼠单胺类递质和炎症的影响

目的:通过条件性恐惧模型大鼠来评估跑台运动的抗创伤后应激障碍(PTSD)作用,并探讨运动对PTSD影响的可能分子机制。方法:将50只雄性SD大鼠随机分为对照组(control)、条件恐惧模型组(FC)、消退组(EXT)、消退+运动组(EXT+EX)和消退+舍曲林组(EXT+SER)。通过条件恐惧消退保持测试来测试情境恐惧记忆;通过自发活动测试和高架迷宫测试来测试大鼠焦虑样行为;利用酶联免疫法(ELISA)检测大鼠海马肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)水平;利用高效液相色谱(HPLC)技术测定大鼠海马和前额叶皮质中单胺类神经递质及其代谢产物水平。结果:FC组大鼠的僵住时间增加(P 0.001),进入开臂次数百分比(P 0.01)和开臂时间百分比(P 0.001)减少,而运动和舍曲林均逆转了这些行为缺陷(P 0.05)。FC大鼠海马中TNF-α水平(P 0.05)和IL-1β水平(P 0.05)显著升高,FC大鼠海马和皮层中的去甲肾上腺素(NE)水平(P 0.05)以及多巴胺(DA)水平(P 0.05)显著升高。而舍曲林给药显著降低了海马中TNF-α水平(P 0.05),跑台运动和舍曲林给药显著均显著降低了海马中IL-1β水平(运动:P 0.05;舍曲林:P 0.01)和5-羟色胺(5-HT)代谢产物5-羟吲哚乙酸(5-HIAA)水平(P 0.05),同时显著降低了皮层中NE水平(P 0.01)和DA水平(P 0.05)。结论:跑台运动改善了条件恐惧模型大鼠的情境恐惧记忆和焦虑样行为。单胺类神经递质NE和DA以及炎性因子TNF-α和IL-1β可能参与了跑台运动对PTSD的缓解作用。     

卵巢切除对大鼠子宫雌激素受体亚型的影响

雌激素在生长、分化及雌性生殖功能中扮演重要角色.雌激素的作用是由靶细胞内雌激素受体(estrogen receptor,ER)所介导的,ER是类固醇受体超家族成员之一,它是通过与雌激素受体反应元件结合调控基因转录,而在靶细胞中发挥功能的.     

米氮平配伍雌激素与单纯雌激素补充治疗更年期抑郁症的对照研究

目的探讨米氮平与雌激素联合治疗更年期抑郁症的疗效.方法更年期抑郁症患者60例,随机分为两组,分别使用米氮平联合雌激素（倍美丽）治疗（观察组）,单用雌激素（倍美丽）治疗（对照组）,疗程8周.采用汉密顿抑郁量表（HAMD）评定疗效.结果治疗8周后观察组HAND评分显著低于对照组;观察组显效率显著高于对照组.结论米氮平联合雌激素治疗更年期抑郁症起效快,效果好.     

雌激素受体β在小鼠及猪卵巢中表达的比较研究

目的 研究雌激素受体β(ERβ)在小鼠和猪卵巢中的表达差异. 方法通过免疫组织化学的方法,研究ERβ在小鼠及猪卵巢中不同类型细胞中的表达量和细胞内的定位情况. 结果在小鼠卵巢中,ERβ在黄体和各级生长卵泡的卵母细胞质中均有大量表达,在各级卵泡的颗粒细胞和膜细胞中表达量很少,生殖上皮和间质细胞中没有检测到表达信号.猪卵巢中ERβ强表达于各级卵泡的膜细胞中,各种直径的腔前卵泡的颗粒细胞中也有表达,但有腔卵泡颗粒细胞有的表达,有的则未见表达,间质细胞和生殖上皮中无明显表达. 结论 ERβ在小鼠和猪卵巢中的表达和定位差异很大.表明ERβ在小鼠和猪卵巢中的作用也可能不同.     

雌激素受体抑制剂对小鼠囊胚形成的影响

目的观察雌激素受体（estrogen receptor,ER）抑制剂对昆明（Kunming,KM）小鼠体外囊胚形成的影响,为进一步探讨雌激素受体在小鼠囊胚形成中的作用和机制提供实验依据。方法以空白KSOM培养液为对照组,KSOM中添加浓度为1、2、3、4、5p~mol／L的ER仪抑制剂MPP,以及浓度为1、10、50、100μmol／L的ERB抑制剂PHTPP为实验组,收集KM小鼠8．细胞胚进行体外连续培养,计数各组发育至桑葚胚和囊胚两个阶段的数目,比较各组发育到囊胚的比率。结果添加浓度为1~mol／L、2Ixmol／L的MPP实验组囊胚发育率分别为86．36％、86．40％,与对照组（82．26％）相比无明显差别（P〉O．05）,而添加浓度为3μmol／L、4μmol／L、5μmol／L的MPP实验组囊胚发育率分别为46．58％、7．50％、0,发育率显著下降（P〈0．01）。添加浓度为1μmol／L、10μmol／L、50~mol／L的PHTPP实验组囊胚发育率分别为82．61％、87．04％、83．98％,与对照组（82．09％）比较没有明显差异（P〉O．05）,而添加高浓度（100μmol／L）的PHrrPP实验组囊胚发育率（51．39％）较明显下降。结论低浓度（5μmol／L）的ERo的抑制剂MPP可完全抑制KM小鼠8一细胞胚体外发育到囊胚,而50~mol／L的ERB抑制剂PHTPP对KM小鼠囊胚形成无明显影响,高浓度（100μmol／L）的PHTPP才可部分抑制囊胚形成。提示在小鼠囊胚形成中,ERα起着更为主要的作用。     

葛根素对围绝经期抑郁症模型小鼠的神经保护作用及机制研究

目的:探讨葛根素对围绝经期抑郁症模型小鼠行为影响及神经生物学机制。方法:清洁级昆明小鼠饲养1周以适应环境。旷场实验(OFT)筛选后,随机分为假手术组、围绝经期抑郁症模型组、氟西汀组(FLU,3mg/kg)、雌激素组(E,0.15 mg/kg)和葛根素组(92 mg/kg),每组10只。采用小鼠双侧卵巢切除(OVX)联合慢性不可预知性温和应激(CUMS)法建立围绝经期抑郁症动物模型。各给药组于每日应激前1 h灌胃给药,其余各组给予等体积生理盐水,共计21 d。观察小鼠动情周期变化,测定行为学变化,观察海马组织形态学改变,测定脑组织单胺类递质含量,测定海马组织c AMP反应元件结合蛋白(CREB)-脑源性神经营养因子(BDNF)信号通路主要蛋白表达。结果:与假手术组比较,各OVX组小鼠连续7 d内连续监测未见动情周期变化,证明去势成功;围绝经期抑郁症模型组小鼠呈现抑郁样行为,表现为自发活动及探索行为减少、行为绝望时间增加、体质量增长缓慢(P0.01或P0.05),海马神经元损伤、萎缩、数量减少、尼氏体减少及早期凋亡变化,脑组织五羟色胺(5-HT)、去甲肾上腺素(NE)及多巴胺(DA)含量减少(P0.01);海马组织CREB-1、磷酸化c AMP反应元件结合蛋白(p-CREB-1)、BDNF及酪氨酸蛋白激酶B(Trk B)蛋白表达减少(P0.01)。与围绝经期抑郁症模型组比较,给予葛根素(Pue)治疗后,可改善围绝经期抑郁症模型小鼠抑郁样行为,表现为增加自发活动及探索行为、减少行为绝望时间、体质量增加迅速(P0.01或P0.05);减轻海马组织神经元损伤及凋亡细胞,增加脑组织5-HT、NE及DA含量(P0.01),增加海马组织CREB-1、p-CREB、BDNF及Trk B蛋白表达(P0.01),差异具有统计学意义。结论:采用小鼠双侧OVX联合CUMS可成功制备围绝经期抑郁症动物模型。葛根素具有神经保护及抗围绝经期抑郁症作用,其机制主要通过减轻海马神经元损伤、抑制神经元早期凋亡、增加脑组织单胺类递质含量及上调脑组织CREB-BDNF信号通路主要蛋白表达而发挥的。     

Macamide B对缺氧缺血性脑损伤新生小鼠脑中雌激素受体表达的影响

目的:观察玛卡酰胺B(Macamide B)对缺氧缺血性脑损伤(HIBD)新生小鼠脑组织中雌激素受体α(ER α)和雌激素受体β(ER β)表达的影响。方法:选择出生7 d C57BL/6小鼠30只并将其分为3组:假手术组(sham)、缺氧缺血性脑损伤组(HIBD)、玛卡酰胺B处理组(Macamide B+HIBD),每组10只。采用Rice法建立新生小鼠HIBD模型,采用免疫荧光组织化学染色对ER α和ER β在新生小鼠HIBD脑内的细胞定位进行分析,Western Blot方法检测Macamide B对新生鼠HIBD脑内ER α和ER β表达的影响。结果:ER α和ER β在HIBD新生小鼠脑内神经元和星形胶质细胞中均有表达;与sham组相比,HIBD组ER α和ER β的表达量均明显降低;与HIBD组相比,Macamide B可显著上调ER α和ER β的表达。结论:Macamide B可通过上调新生小鼠脑内ER α和ER β表达表达水平对HIBD发挥神经保护作用。     

Effect of estrogen on the uterus of various strains of mice

Laboratory of Carcinogens, Research Institute of Carcinogenesis, All-Union Oncologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. N. Trapeznikov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 112, No. 12, pp. 631–633, December, 1991.     

Alterations in the phenotype and function of immune cells in ovariectomy-induced osteopenic mice

BACKGROUND: Within the last few years, much evidence has been presented on the involvement of the immune system in certain types of bone loss, such as activated T cells in rheumatoid arthritis and in periodontitis. Estrogen deficiency induces bone loss; however, how this deficiency affects the immune system has not been sufficiently studied. METHODS: To evaluate the effects of estrogen withdrawal on the status and functionality of the immune system, mice were ovariectomized or sham-operated, and 5 weeks after surgery, when osteopenia had developed, several parameters were analysed in spleen and in bone marrow. We analysed bone turnover, cell phenotype by flow cytometry, cell function by cell proliferation assays, and the expression of several genes related to the process. RESULTS: Five weeks after ovariectomy, augmented osteoclastogenesis persisted in the bone marrow. In addition, the ovariectomized mice had more B-cells and CD3+ T-cells expressing the receptor activator of NF-kappaB ligand (CD3+/RANKL+). The ovariectomized mice had lower serum alkaline phosphatase activity, a normal amount of T cells, lower percentages of CD11b+ and CD51+ cells in the bone marrow, and a lower serum interferon-gamma level compared with sham-operated controls. CONCLUSIONS: The data suggest that, 5 weeks after ovariectomy, bone turnover remains imbalanced, with increased osteoclastogenesis and a decreased rate of bone formation. Moreover, there is an increase in B-cell formation, with normal and decreased percentages of T cells and myelomonocytic cells (CD11b+), respectively, in the bone marrow. Decreased serum interferon-gamma levels could be involved in the increased osteoclastogenesis found in the present work.     

Analysis of the effect of estrogen/androgen perturbation on penile development in transgenic and diethylstilbestrol‐Treated mice

Because both androgens and estrogens have been implicated in penile morphogenesis, we evaluated penile morphology in transgenic mice with known imbalance of androgen and estrogen signaling using scanning electron microscopy (SEM), histology, and immunohistochemistry of androgen and estrogen receptors α/β. Penises of adult wild‐type, estrogen receptor‐α knockout (αERKO), estrogen receptor‐β knockout (βERKO), aromatase knockout (Arom‐KO), and aromatase overexpression (Arom+) mice were evaluated, as well as adult mice treated with diethylstilbestrol (DES) from birth to day 10. Adult penises were examined because the adult pattern is the endpoint of development. The urethral orifice is formed by fusion of the MUMP (male urogenital mating protuberance) with the MUMP ridge, which consists of several processes fused to each other and to the MUMP. Similarly, the internal prepuce is completed ventrally by fusion of a ventral cleft. In adult murine penises the stromal processes that form the MUMP ridge are separated from their neighbors by clefts. αERKO, βERKO, and Arom‐KO mice have penises with a MUMP ridge clefting pattern similar to that of wild‐type mice. In contrast, Arom+ mice and neonatally DES‐treated mice exhibit profound malformations of the MUMP, MUMP ridge clefting pattern, and internal prepuce. Abnormalities observed in Arom+ and neonatally DES‐treated mice correlate with the expression of estrogen receptor‐beta (ERβ) in the affected structures. This study demonstrates that formation of the urethal orifice and internal prepuce is due to fusion of separate epithelial‐surfaced mesenchymal elements, a process dependent upon both androgen and estrogen signaling, in which ERβ signaling is strongly implicated. Anat Rec, 296:1127–1141, 2013. © 2013 Wiley Periodicals, Inc.     

抑郁小鼠模型脑突触体中突触蛋白表达的变化

目的:研究抑郁症发病过程中突触蛋白表达的变化。方法:小鼠随机分为对照组和造模组,使用社会失败模型对造模组小鼠造模,之后将其分为对抑郁症敏感组及对抑郁症不敏感组。将以上3组小鼠分别断头取前额叶皮层、海马、纹状体并提取突触小体,通过免疫印迹检测突触小体内突触后致密蛋白95(PSD-95)、桥尾蛋白(gephyrin)及突触蛋白1(synapsin-1)蛋白表达的变化。结果:与对照组相比,在前额叶皮层,对抑郁症敏感组及对抑郁症不敏感组小鼠的PSD-95水平下降,而对抑郁症敏感组小鼠的synapsin-1表达增多;在海马,对抑郁症敏感组及对抑郁症不敏感组小鼠的PSD-95水平下降;在纹状体,对抑郁症不敏感的小鼠gephyrin、synapsin-1蛋白表达显著增加。结论:在抑郁症发病中突触体内兴奋性及抑制性蛋白发生了变化,这些可能与突触功能的紊乱相关。     

雌激素对去势大鼠髁突软骨雌激素受体基因表达的影响

目的探讨不同浓度的雌二醇(E2)对去势大鼠髁突软骨雌激素受体(ER)基因ERα和ERβ表达的影响。方法将SD大鼠分为对照组(control)、假手术组(sham)、去势组(OVX)、及时和延迟替代治疗组(OVX/17β-E2),每天静脉注射不同浓度雌二醇(5×10、5×10~2和5×10~3μg/kg),阴道上皮脱落细胞涂片监测雌激素水平,用Western blot和real-time PCR法检测大鼠颞下颌关节髁突软骨ERα和ERβ的蛋白及mRNA表达。结果去势大鼠阴道上皮细胞MI指数180/12/8,表明雌激素水平低下(P0.05)。及时补给生理浓度雌二醇(5×10~2μg/kg)能维持髁突软骨ER于正常范围,延迟补给生理浓度雌二醇不能恢复或改善ER表达;及时和延迟补给高浓度(5×10~3μg/kg)及低浓度(50μg/kg)E2均抑制ER蛋白和mRNA合成(P0.05),且高浓度效应显著。结论及时补充合理浓度的雌激素对去势大鼠颞下颌关节的正常生理功能具有重要意义。     

小鼠室旁核内雌激素受体与催产素、加压素表达的免疫组化研究

目的研究雌激素的核受体ER-α和ER-β以及催产素、加压素在成年雌性小鼠下丘脑室旁核内的表达。方法采用硫酸镍铵增强显色的免疫组化SP法检测ER-α、ER-β、催产素和加压素在室旁核内的表达。结果雌激素的两种核受体在室旁核内都有表达,但是以ER-β为主（P〈0.001）,其免疫阳性产物均在细胞核内,未见核外免疫阳性反应。催产素的免疫阳性产物主要在细胞核周围的胞浆即核周质内,而加压素的免疫阳性物质除了在核周质内有很强的反应外,在突起内也可见很强的免疫反应。ER-α的免疫阳性胞体主要在大细胞部内侧,而ER-β、催产素和加压素的免疫阳性胞体主要在背侧帽部或大细胞部的外侧。结论室旁核内两种雌激素受体可能都参与了对催产素和加压素的调节,但ER-β可能发挥了主要的调节作用。     

Postmenopausal effects of intrastriatal estrogen on catalepsy and pallidal electroencephalogram in an animal model of Parkinson's disease

The main objective was to test the preventive and treatment effects of central injection of estrogen (ES) on muscular rigidity and pallidal EEG in menopausal rats' model of Parkinson's disease (PD). We hypothesized that intrastriatal pretreatment and post-lesion treatment by estrogen improve the pallidal local EEG and muscular stiffness in animal model of menopause with PD. Forty-eight female Wistar rats (300-350 g) were ovariectomized (OVX) and divided into two main groups: Non-pretreated subgroups; sham (S), lesioned (L), post-lesion treated (LT) and pretreated subgroups; pretreated (Pt), pretreated and then lesioned (PtL), pretreated and post-lesion treated (PtLT). Pallidal local EEG was recorded by a bipolar recording electrode and muscle stiffness was scored by Dekundy's test in freely moving rats. Muscle stiffness and pallidal local EEG were indicated as main outcome measures. In pretreated group the local pallidal EEG was decreased in sham-operated rats compared with non-pretreated group (P<0.01), and SNc lesioning decreased EEG in the non-pretreated (P<0.01), while it increased the EEG in the pretreated group (P<0.01). In both groups administration of ES restore the EEG to the respective sham-operated group (P<0.01). Regarding muscle stiffness, it increased after SNc lesioning in both pretreated and non-pretreated groups and ES administration decreased the rigidity significantly (P<0.05, P<0.001 respectively). However, the lesion-induced rigidity in pretreated groups was significantly less than non-pretreated groups (P<0.05). Because of its modulatory effect estrogen may protect dopaminergic neurons from injury and may interfere with the uptake of 6-hydroxydopamine (6-OHDA) into the nigral dopaminergic neurons or alter dopamine release and uptake in remaining neurons.     

Effects of a7nAChR agonist on the tissue estrogen receptor expression of castrated rats

Osteoporosis is one common disease in postmenopausal women due to depressed estrogen level. It has been known that inflammatory factors are involved in osteoporosis pathogenesis. One regulator of inflammatory cascade reaction, a7-nicotinic acetylcholine receptor (a7nAChR), therefore, may exert certain role in osteoporosis. This study thus investigated this question on an osteoporosis rat model after castration. Rats were firstly castrated to induce osteoporosis, and then received a7nAChR agonist (PNU-282987), diethylstilbestrol or saline via intraperitoneal injection. After 6 or 12 weeks, bone samples were collected for counting osteoblast number, bone density and estrogen receptor (ERα and ERβ) expression, in addition to the serum laboratory of inflammatory factors. Bone density, osteoclast number, ERα and ERβ expression level were significantly depressed in model group, and were remarkable potentiated in the drug treatment group (P<0.05). The levels of BGP and PTH in drug treatment group were decreased compared to diethylstilbestrol group, while E2 and IGF-1 showed up-regulation. Agonist of a7nAChR can up-regulate estrogen receptor expression and may prevent the occurrence and development of osteoporosis.