雌激素通过miR-146a影响间充质干细胞的免疫调节功能

本研究用混合酶法分离培养人脐带间充质干细胞(MSC),检测在不同浓度17β-雌二醇(E2)作用下MSC中MicroR-NA的表达情况。进一步通过转染使MSC高表达miR-146a,并分析转染后MSC的表型、功能及相关因子的表达变化。结果显示,E2处理的MSC中IFN-β的表达没有发生明显变化,STAT1与IDO1的表达显著下调。同时,E2处理能够上调MSC对miR-146a的表达,降低其对miR-23b和miR-21的表达。miR-146a转染不影响MSC的表型,而显著降低STAT1和IDO的表达。提示E2可能通过影响MSC内miR-146a的表达改变其免疫调节功能。     

Motherhood alters the cellular response to estrogens in the hippocampus later in life

Although controversial, estrogen replacement therapy has been implicated as a possible therapeutic agent for ameliorating age-related cognitive decline in postmenopausal women. We have shown previously that different types of estrogen promote hippocampal neurogenesis in a dose-dependent manner in young adult female rats. However, previous studies have not found a beneficial effect of 17β-estradiol in middle-aged female rats. The aim of the present study was to determine the acute effects of 17β-estradiol, 17α-estradiol, and estrone on hippocampal cell proliferation in middle-aged ovariectomized female rats and to determine whether effects are dependent on previous reproductive experience. Middle-aged multiparous female rats or age-matched virgin female rats were injected subcutaneously with vehicle or 10 μg dose of 17β-estradiol, 17α-estradiol, or estrone, and then given BrdU 30 min later and perfused 24 h later to assess cell proliferation. All estrogens significantly upregulated cell proliferation in the hippocampus in middle-aged multiparous females but none of the estrogens upregulated cell proliferation in the middle-aged virgins. Therefore, previous reproductive experience may make the older brain more responsive to estrogens later in life. We also found that 17α-estradiol upregulated cell proliferation to a greater degree than the other estrogens in the multiparous females. Together these findings may lead to the development of new therapeutic advances in the treatment of symptoms associated with menopause in women.     

Acute effect of intranasal estrogen on cerebral and cerebellar perfusion in postmenopausal women

OBJECTIVES: Estrogen action in the brain influences many neurochemical processes. The aim of the study was to evaluate the acute effect of intranasal 17beta-estradiol on cerebral and cerebellar perfusion in postmenopausal women. METHODS: The study group included 24 healthy postmenopausal women who had been in natural menopause for at least 1 year (mean age: 47.38+/-5.9 years). We conducted an experimental, randomized, placebo-controlled, cross-over, double-blind study. Cerebral and cerebellar perfusion was measured after placebo (saline serum physiologic) or intranasal 17beta-estradiol administration by Single Photon Emission Computed Tomography (SPECT) using technetium-99m-hexamethylpropylene amine oxime (Tc99m-HMPAO). Regions of interest (ROIs) were drawn manually. Cerebral and cerebellar perfusions were calculated for each ROI using average number of counts per pixel. Semiquantitative analysis was performed in bilateral frontal, temporal, parietal and occipital lobes, thalamus, putamen, hippocampus, amygdala, caudate nuclei, cerebellar region, anterior/posterior of cingulate gyrus and pons. RESULTS: After intranasal 17beta-estradiol administration, SPECT study revealed significant increases in cerebral and cerebellar perfusion compared to placebo measurements in all studied slices (p<0.05). There was a positive correlation between serum estrogen levels after 17beta-estradiol and cerebral and cerebellar perfusion. CONCLUSIONS: Administration of single dose intranasal 17beta-estradiol increases cerebral and cerebellar perfusions in healthy postmenopausal women.     

17β-雌二醇调节小鼠淋巴结树突状细胞的表型和吞噬功能

为探索17β-雌二醇(E2)对免疫应答能力的调节是否与树突状细胞(DC)的成熟和功能相关,用Metrizamide密度梯度离心方法分离BALB/c小鼠淋巴结内的淋巴细胞得到DC,不同浓度的E2处理DC 24 h后,使用PE标记的单抗CD11c和FITC标记的单抗MHC I/MHC II/CD40/CD54/CD80/CD86、IL-10/TNF-α以及右旋糖酐(dextran)双标DC,流式细胞仪分别检测其表型和胞内细胞因子的表达及吞噬能力的变化。结果显示,不同浓度的E2处理DC 24 h后,MHC分子、黏附分子CD54和共刺激分子CD86与对照组相比显著提高,而CD40显著降低,但CD80的表达无明显改变。胞内细胞因子IL-10和TNF-α的表达水平随E2的处理而显著增加,其中TNF-α的变化呈剂量依赖性。与对照组相比,DC的吞噬能力随E2的处理浓度增加而显著降低。以上结果表明,E2能够改变DC的成熟和功能,这提示E2对机体的免疫应答的影响可能通过DC的改变而改变。     

雌激素及其拮抗剂对大鼠脂代谢和血管活性肽水平的影响

以假手术大鼠（Sham）作为对照组,切除大鼠双侧卵巢（OVX）后,血浆17β-雌二醇（17β-E2）水平下降48％。体重增加17％（P＜0．001）,血清总胆固醇（TC）含量升高约50％（P＜0．001）,甘油三酯（TG）含量降低30％（P＜0．01）。OVX后进行雌激素替代疗法（ERT）各组（ERT1-3）的TC和TG的含量分别下降（P＞0．05）到和升高（P＜0．01）至高于对照组的水平。给于雌激素受体拮抗剂──它莫西芬（TAM）的各组（TAM,OVX＋TAM,ERT3＋TAM）中,血清TC和TG含量均低于对照组（P＜0．01～P＜0．001）。此外,OVX组血浆降钙素基因相关肽（CGRP）和内皮素水平也分别降低为对照组的69％和89％（P＜0．05）。提示卵巢激素主要是雌激素通过其受体介导参与大鼠脂代谢和体重的调节,ERT和TAM对OVX大鼠血清TC浓度及体重的调节产生有利影响。OVX大鼠血浆CGRP和内皮素水平的调节可能与雌激素水平变化有关。     

17β-雌二醇通过microRNA-29b调节NK细胞干扰素γ的分泌

目的:研究17β-雌二醇(E2)对人自然杀伤细胞株NK92中干扰素γ(IFN-γ)分泌的影响及其可能的机制。方法:用E2和poly I:C处理NK92细胞,分别在4、8、12和24小时后用实时定量PCR、酶联免疫吸附法和流式细胞术测定NK92细胞中IFN-γ的表达情况,用实时定量PCR检测microRNA-29b(miR-29b)的变化。进一步,构建含有IFN-γ3’UTR的pEGFP-C1质粒,并转染pre-29b和pEGFP-IFN-γ3’UTR质粒至HEK293A细胞中,用流式细胞术检测绿色荧光蛋白(GFP)的变化。结果:E2抑制NK92细胞的IFN-γ分泌和上调miR-29b的表达;miR-29b能够直接靶向调节IFN-γ3’UTR mRNA。结论:E2可能通过调节NK92的miR-29b而影响IFN-γ的分泌水平,进而影响其免疫功能。     

Ethanol withdrawal posttranslationally decreases the activity of cytochrome c oxidase in an estrogen reversible manner

Cytochrome c oxidase (COX) is a key mitochondrial enzyme that catalyzes electron transfer at the terminal stage of respiratory chain and is composed of multisubunits. We hypothesize that ethanol withdrawal (EW) impairs the activity of COX and estrogen deprivation exacerbates this problem. Five-month-old ovariectomized rats with or without 17beta-estradiol (E2) replacement received a control dextrin or a liquid ethanol diet (6.5%, 5 weeks). They were then sacrificed either during ethanol exposure or at 24h of EW (EW group). Mitochondria of the cerebellum and cortex were processed to measure the activities of total COX, COX subunit I, and IV. The effects of EW and E2 on the protein levels of these subunits were also assessed using an immunoblotting method. As compared to the control dextrin and ethanol exposure, EW decreased the activities of total COX, COX I, and COX IV. E2 treatment prevented the effects of EW on the activities of total COX and COX IV but not COX I. Neither EW nor E2 altered the protein levels of the subunits. These findings suggest that a counteracting relationship exists between the effects of EW and E2 on the activity of COX in a subunit specific manner.     

Synaptic plasticity in the substantia gelatinosa in a model of chronic neuropathic pain

Chronic neuropathic pain (CNP) is common after peripheral nerve injuries (PNI), but is rather refractory to available anti-pain medication. Advances in neuropathic pain research have identified cellular and molecular cues triggering the onset of neuropathic pain, but the mechanisms responsible for maintenance of chronic pain states are largely unknown. Structural changes such as sprouting of injured A-fibres into the substantia gelatinosa of the dorsal horn in the spinal cord have been proposed to relate to neuropathic pain in partial PNI models. Structural changes in central pain networks may also underlie the more persistent CNP following complete sectioning of a peripheral nerve, because this type of injury results in continuous and spontaneous afferent input to the spinal cord, which can trigger central sensitization. In the present study, the left sciatic nerve was completely sectioned and a 1-cm segment was removed to maintain a chronic pathology, whereas the right sciatic nerve was left intact. Mechanical allodynia was measured up to 84 days after injury, after which synaptic changes were studied in the lumbar substantia gelatinosa. The numbers of larger sized synaptophysin-immunoreactive presynaptic boutons were found to be increased in the substantia gelatinosa ipsilateral to the nerve injury. From these data we conclude that structural synaptic changes within the substantia gelatinosa are present months after complete nerve injury and that this plasticity may be involved in maintaining neuropathic pain states.     

Sustained acceleration of colonic transit following chronic homotypic stress in oxytocin knockout mice

A sex difference has been reported in the responsiveness of the vomeronasal (VN) system to pheromones. In the present study, to clarify a direct and acute influence of 17β-estradiol (E2) on the accessory olfactory bulb (AOB) neurons, we investigated the effect of E2 on dendritic spines in cultured AOB cells derived from male and female neonatal rats. After 17-18 days in vitro (DIV), cultured AOB cells were transfected with GFP expression vectors. At 21-23 DIV, cells were treated with E2, and time-lapse images of transfected AOB neurons identified as granule cells were taken under a confocal laser scanning microscope for 3h. The dendritic spine head area of granule cells was quantitatively evaluated, and spine heads were classified into larger (≥ 1 μm2) and smaller (<1 μm2) ones before E2-treatment (0 h). In cultured cells derived from both sexes, the larger spines were not significantly changed at 1, 2 and 3 h after E2-treatment. In contrast, E2-treatment significantly enlarged the head area of the smaller spines of granule cells derived from the female, whereas E2 did not cause any significant effects on those from the male. Our results provide evidence for the sexually-dimorphic effect of E2 on spine development in AOB granule cells.     

Effect of OS-0689, a novel SERM, on periarterial nerve function in tail arteries of ovariectomized rats

OBJECTIVE: We have previously shown that OS-0689 attenuates the rise in tail skin temperature of ovariectomized rats, which is believed to be relevant to human symptoms of hot flush. In this study, we elucidate the mechanism underlying the ameliorating effects of OS-0689 on elevated tail skin temperature. METHODS: Female Sprague-Dawley rats were ovariectomized and orally treated with OS-0544 (1 mg/kg), OS-0689 (3 mg/kg; (+)-enantiomer of OS-0544) or 17beta-estradiol (3 mg/kg; E2) for 1 week. At 1, 3 or 6 weeks after ovariectomy, the vasoconstrictions and vasorelaxations induced by periarterial nerve stimulation (PNS), l-noradrenaline (NA), and rat calcitonin gene-related peptide (CGRP) in isolated tail arteries were compared between OVX and sham-operated rats. RESULTS: Three weeks after ovariectomy, vasoconstrictions in response to PNS and NA in the arteries of OVX rats were markedly less than those in the arteries of sham-operated rats. However, at 1 and 6 weeks after ovariectomy the stimuli-induced vasoconstrictions in the arteries of OVX rats were greater than those of sham-operated rats. Moreover, NA reactivity was not attenuated in the mesenteric arteries at 3 weeks after ovariectomy. OS-0544, OS-0689 and E2 prevented the decrease in vasoconstrictions in the tail arteries. Vasorelaxations in response to PNS and rat CGRP were significantly greater in the arteries of OVX rats than in those of the sham-operated rats. OS-0689 inhibited the increase in vasorelaxation induced by both stimuli, whereas E2 had no effects. CONCLUSIONS: Ovariectomy not only decreases adrenergic function but also enhances CGRPergic function in rats' tail arteries. OS-0689 improves both impairments and thereby improves on rat hot flush.     

IL-23在慢性乙型肝炎免疫调节中的作用研究

目的 探讨IL-23在慢性乙型肝炎免疫调节中的作用。方法 选取我院收治的32例HBeAg阳性慢性乙型肝炎患者为研究对象,根据ALT水平分为ALT≥120 IU/ml患者16例,ALT＜120 IU/ml患者16例,另外选择我院健康体检中心20例体检者作为健康对照组,采用酶联免疫分析法（ELISA）检测血清IL-23水平,采用流式细胞仪检测Th17细胞百分率。结果 与健康对照组相比,HBeAg阳性慢性乙型肝炎患者血清IL-23表达及外周血Th17细胞百分率均升高,差异有统计学意义（P＜0.05）; ALT≥120 IU/ml患者血清IL-23浓度（394.81±101.84）pg/ml高于ALT＜120IU/ml的（283.69±85.65）pg/ml,ALT≥120 IU/ml患者Th17细胞百分率（3.25±0.70）%高于ALT＜120IU/ml的（2.68±0.61）%,差异均有统计学意义（P＜0.05）;慢性乙型肝炎患者外周血Th17细胞百分率、血清IL-23浓度与ALT程度呈正相关（P均＜0.05）;Th17细胞百分率与血清IL-23浓度呈正相关（P＜0.05）。结论 IL-23可能通过影响Th17细胞的免疫调节参与慢性乙型肝炎患者炎症,为临床提供了新的靶标。     

17β-Estradiol regulates insulin-degrading enzyme expression via an ERβ/PI3-K pathway in hippocampus: Relevance to Alzheimer's prevention

Insulin-degrading enzyme (IDE), an enzyme that primarily degrades insulin, has recently been demonstrated to play a significant role in the catabolism of amyloid β (Aβ) protein in the brain. Reduced IDE expression and/or activity have been associated with the etiology and development of Alzheimer's disease (AD). Using three model systems, the present investigation provides the first documentation indicating that estrogen robustly regulates the expression of IDE in normal, menopausal and early-stage AD brains. In vitro analyses in primary cultures of rat hippocampal neurons revealed that 17β-estradiol (17β-E2) increased IDE in both mRNA and protein levels in a time-dependent manner. Further pharmacological analyses indicated that 17β-E2-induced IDE expression was dependent upon estrogen receptor (ER) β and required activation of phosphatidylinositol 3-kinase (PI3-K). In vivo analyses in adult female rats revealed a brain region-specific responsive profile. Ovariectomy (OVX) induced a significant decline in IDE expression in the hippocampus, which was prevented by 17β-E2. Neither OVX nor 17β-E2 affected IDE expression in the cerebellum. In vivo analyses in triple transgenic AD (3xTg-AD) female mice revealed an inverse correlation between the age-related increase in Aβ load and the decrease in IDE expression in the hippocampal formation. Treatment with 17β-E2 attenuated Aβ accumulation/plaque formation and elevated hippocampal IDE expression in 12-month-old 3xTg-AD OVX mice. Collectively, these findings indicate that 17β-E2 regulates IDE expression in a brain region-specific manner and such a regulatory role in the hippocampus, mediated by an ERβ/PI3-K pathway, could serve as a direct mechanism underlying estrogen-mediated preventative effect against AD when initiated at the onset of menopause.     

Synaptic pathology in Alzheimer's disease: a review of ultrastructural studies

Morphologic studies of the neuropathology in Alzheimer’s disease (AD) have demonstrated significant loss of synaptic connectivity in many regions of the neocortex and hippocampus. The strongest correlation with cognitive decline in AD is with the synaptic density. This article discusses the ultrastructural studies that have documented changes in synaptic numbers in many areas of association cortex and in the hippocampal dentate gyrus molecular layer. Changes in the synaptic complex are discussed as a possible compensatory mechanism in response to synapse loss and a model is proposed to help relate the significance of these synaptic changes. Comparisons are made between results observed with ultrastructural technique and those utilizing immunohistochemistry to assess changes in synaptic pathology. Possible reasons underlying the synaptic neuropathology are discussed.     

雌二醇对大鼠颌下腺GnRH、GH生成的影响

目的　研究雌二醇 (estradiol- 17β,E2 )对颌下腺促性腺激素释放激素 (gonadotropin releasing hor-mone,Gn RH)和生长激素 (growth factor,GH )生成的影响。方法　采用免疫组织化学的 ABC法。结果　雌二醇促进颌下腺 Gn RH的生成而对 GH的产生有抑制作用。结论　雌二醇可能对颌下腺 Gn RH和 GH的合成起重要的调节作用     

Estrogen Regulation of Lactoferrin Expression in Human Endometrium

PROBLEM: Lactoferrin is an iron-binding glycoprotein that has been shown to be overexpressed in human endometrial carcinomas. The purpose of our present study is to investigate the possible role of estradiol in the expression of lactoferrin. METHOD OF STUDY: We investigated 1) serum levels of lactoferrin in five women during normal ovulatory cycles, 2) serum levels of lactoferrin during ten human menopausal gonadotropin induced cycles when estradiol levels are high, and 3) lactoferrin expression in five proliferative and five secretory phase endometrium by immunohistochemical studies. The serum concentrations of lactoferrin were measured by a peroxidase-based enzyme-linked immunosorbent assay. RESULTS: In normal ovulatory cycles, the mean serum lactoferrin concentration during the proliferative phase (0.4013 ± 0.0242 μg/mL) was significantly higher (P<0.02) than in the secretory phase (0.3468 ± 0.0209 μg/mL). In induced cycles, there was gradual increase in lactoferrin levels with increasing estradiol concentrations. Peak lactoferrin levels in induced cycles (0.7495 ± 0.1148 μg/mL) were significantly higher (P<0.003) than the midcycle levels (0.423 ± 0.0424 μg/mL) in normal cycles. Immunohistochemical analysis of the endometrium revealed greater expression of lactoferrin in proliferative endometrium (50.7 ± 13%, range 28–72%) than in secretory endometrium (19.2 ± 4%, range 7–31%). CONCLUSION: These results indicate that estradiol may play a role in the regulation of lactoferrin expression in human endometrium.     

缺氧缺血性脑损伤新生大鼠海马突触体素的表达及当归调控作用

目的探讨脑缺氧缺血对新生大鼠海马齿状回突触体素表达的影响及当归注射对其表达的调控作用。方法取7日龄健康SD新生大鼠33只,随机分为对照组、缺氧组和当归组各11只。缺氧组和当归组新生大鼠在无菌环境下结扎左侧颈总动脉,术后护理2 h后置于三气培养箱持续缺氧2 h,制作新生鼠缺氧缺血性脑损伤（HIBD）模型,对照组仅行假手术,不结扎左侧颈总动脉、不缺氧。术后第8 d开始,缺氧组和对照组大鼠经腹腔注射生理盐水（8ml/kg）,连续7 d;当归组用等量当归注射液（250 g/L）代替生理盐水。于生后第22 d取大鼠脑组织,常规石蜡包埋、经海马切片,行突触体素（SY）免疫组化染色,图像分析海马齿状回SY阳性细胞的积分光密度（IOD）值。结果缺氧组大鼠海马齿状回SY阳性细胞的IOD值较对照组降低,而当归组SY阳性细胞的IOD值较缺氧组增高。结论脑缺氧缺血可降低新生大鼠海马齿状回SY的表达,而当归注射液对缺氧缺血性脑损伤新生大鼠可能具有保护作用。     

Effects of Estrogen Treatment on Aging in the Rat Epididymis

Previous studies from our laboratory demonstrated that estrogen signaling in the testis contributes to maintaining spermatogenesis in adult rats, and that estrogen treatment attenuated the age-associated decline in sperm production. The purpose of this study was to determine if epididymal function is also altered with age, and what effects estrogen treatment may have on the epididymis during aging. We compared untreated rats at 3 and 15 months of age to 18-month-old vehicle-treated and estrogen treated rats. In all four groups, tubule and lumen diameter of the cauda was significantly larger than more proximal regions of the epididymis. In the 3-, 15-, and 18-month-old treated animals, the epithelial cell height of the cauda was significantly shorter than that of more proximal regions. The caput cell height was shorter at 18 months compared to 3 months but this was not seen in estrogen treated animals. Thus, estrogen appears able to prevent some age related changes in epididymal morphology. Sperm transit time through the distal cauda was significantly delayed with aging. Estrogen treatment prevented this delay, indicating that sperm transit through the epididymis is an estrogen regulated function. Differences in estradiol and testosterone concentrations were observed between 3- and 15-month-old animals, but no further differences were noted between treated or untreated animals at 18 months. Interestingly, expression of androgen receptor and estrogen receptor alpha were similar between ages and treatments. Collectively, these results suggest epididymal morphology and function are affected by aging and estrogen treatment. Anat Rec, 302:1447–1457, 2019. © 2018 Wiley Periodicals, Inc.