Long-term effect of bilateral ovariectomy on endothelial function in aortic rings of spontaneously hypertensive rats: role of nitric oxide.

Endothelial dysfunction associated with both menopause and hypertension could be one of the possible explanations for increased cardiovascular morbidity and mortality in hypertensive postmenopausal women. The aim of the present study was to investigate the long-term effect of menopause (bilateral ovariectomy) on endothelial function in isolated aortic rings of spontaneously hypertensive rats (SHR). Aortic rings were suspended in organ chambers filled with physiological salt solution (95% O2, 5% CO2, 37 degrees C), and isometric tension was measured. In studies designed to assess the tone-related release of nitric oxide (NO) from phenylephrine-precontracted aortic rings, we found that vasoconstriction induced by L-NAME was greater in aortic rings from sham-ovariectomized SHR (SHAM SHR) than in those obtained from ovariectomized SHR (OVX SHR). Concentration-related relaxant responses to superoxide dismutase were significantly greater in the SHAM SHR than in the OVX SHR. In contrast, receptor-mediated release of NO was not altered by ovariectomy, as deduced from acetylcholine (ACh) concentration-responses curves. Responses to the exogenous NO donor sodium nitroprusside (SNP) were also identical in both ovariectomized and sham-ovariectomized groups, ruling out differences in smooth muscle reactivity to NO. These results show that NO release is impaired in OVX SHR, an animal model of simultaneous hypertension and menopause.     

替勃龙联合更安宁对雌性去势大鼠血管内皮细胞的保护作用

目的:观察替勃龙联合补肾中药(更安宁)在雌性去势大鼠加高脂饮食状态下,对血管内皮细胞的保护作用。方法:9月龄雌性Sprague-Dawley大鼠50只,随机分为假手术组(SHAM)、卵巢切除组(OVX)、卵巢切除后药物联合治疗组(OVX+E)、高脂血症组(OVX+HC)、高脂血症药物联合治疗组(OVX+HC+E)。在卵巢切除术后14d起,OVX组、OVX+HC组给予生理盐水灌胃,OVX+E组、OVX+HC+E组予替勃龙+补肾中药更安宁溶液灌胃,OVX+HC组、OVX+HC+E组同时喂饲高脂饮食。12周后处死大鼠,测定血浆雌二醇(E2)、高密度脂蛋白(HDL-c)、低密度脂蛋白(LDL-c)、胆固醇(Tch)、甘油三脂(TG)、一氧化氮(NO)、血管性假性血友病因子(vWF),在光镜及电镜下观察主动脉管壁结构的改变。结果:OVX+E组与OVX组相比,OVX+HC+E组与OVX+HC组相比,均表现Tch、LDL-c明显降低,HDL-c明显升高,血浆NO含量增加;OVX组存在轻度动脉粥样硬化坏死,OVX+HC组动脉管壁损伤严重,伴随vWF明显升高;OVX+E组与OVX+HC+E组动脉管壁结构正常。结论:替勃龙联合补肾中药更安宁治疗能保护血管内皮细胞,对抗高脂血症对动脉管壁的损伤。     

Ovariectomy exacerbates oxidative stress and cardiopathy induced by adriamycin.

Ovarian hormone depletion in ovariectomized experimental animals is a useful model with which to study the physiopathological consequences of menopause in women. It has been suggested that menopause is a risk factor for the induction of several cardiovascular disorders. In the present study we analyzed the effects of ovarian hormone depletion by ovariectomy (OVX) in a model of oxidative stress and cardiopathy induced by adriamycin (AD). To evaluate these effects, we measured parameters related to cardiac damage (creatinine kinase, lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase) and oxidative stress (malondialdehyde, catalase, superoxide dismutase, glutathione peroxidase, reduced glutathione, nitric oxide and carbonyl proteins) in cardiac tissue and erythrocytes. OVX was found to alter all markers of oxidative stress and cell damage in cardiac tissue. Similarly, the OVX-derived loss of ovarian hormones enhanced cardiac damage and oxidative stress induced by AD. Our results suggest that antioxidant status in cardiac tissue and erythrocytes is seriously compromised by OVX during the cardiomyopathy induced by AD in experimental animals. In conclusion, the absence of hormones caused by OVX or menopause may induce or accelerate pre-existing cardiovascular dysfunctions.     

17β-雌二醇对去势大鼠神经系统的作用

目的：研究补充外源性雌激素对去势大鼠海马和皮质组织中核转录因子2/血红素加氧酶1（Nrf2/HO-1）、沉默信息调节因子相关酶1（SIRT1）表达含量及大鼠海马和皮质区域神经元排列形态和数量的影响,探讨雌激素对神经系统的作用。方法：选取清洁级3个月龄Sprague-Dawley（SD）雌性大鼠40只,随机分为4组。空白组（CON组）、假手术组（SHAM组）、去势对照组（OVX组）、去势实验组（OVX+E2组）,每组10只。OVX+E2组给予17β-雌二醇（E2）灌胃,其余3组予生理盐水灌胃。16周后测定各组大鼠海马和皮质组织中的Nrf2、HO-1、SIRT1表达量,同时对大脑皮质及海马CA1区域神经元行形态学检查。结果：①OVX组大鼠海马及皮质中Nrf2、HO-1水平高于SHAM组,SIRT1水平低于SHAM组;OVX+E2组大鼠海马及皮质中Nrf2、SIRT1水平高于OVX组,HO-1水平则低于OVX组;②OVX+E2组海马及皮质组织中神经元数量及排列情况与SHAM组无明显差异,而OVX组海马及皮质尼氏体数量减少,神经元排列紊乱。结论：补充外源性雌激素可下调去势大鼠海马及皮质中HO-1表达,上调Nrf2、SIRT1表达,维持神经元数量及排列结构,从而保护神经系统。     

替勃龙上调卵巢切除大鼠腰椎护骨素mRNA的表达

目的:建立绝经后骨质疏松(postmenopausalosteoporosis,PMOP)的大鼠模型,研究替勃龙对卵巢切除大鼠腰椎组织中护骨素(OPG)基因mRNA表达水平的影响,探讨其预防和治疗PMOP的作用机制。方法:6月龄未交配健康雌性SD大鼠40只,随机分为SHAM组,OVX组,OVX+戊酸雌二醇(E2)组和OVX+替勃龙(tibolone,TIB)组。灌胃13周后处死动物,第四腰椎行骨组织形态计量指标测定,第二腰椎采用RT-PCR方法,对OPGmRNA表达水平进行检测。结果:OVX组大鼠较SHAM组TBV%显著下降;OSV%明显升高(P<0·05);E2和TIB均可使OVX大鼠的TBV%明显升高,OSV%明显下降;OPGmRNA表达水平在OVX大鼠组织中下调,与SHAM组相比有显著性差异(P<0.05),E2和TIB均可上调OVX大鼠骨组织OPGmRNA表达水平(P<0.05)。结论:E2和TIB均通过抑制骨转换预防和治疗PMOP;PMOP的发生与OPG有关,TIB和E2一样,其抗骨吸收效应很可能是通过OPG介导的。     

Mechanosensitive Vaginal Epithelial Adenosine Triphosphate Release and Pannexin 1 Channels in Healthy,in Type 1 Diabetic,and in Surgically Castrated Female Mice

BackgroundDistension of hollow organs is known to release adenosine triphosphate (ATP) from the lining epithelium, which triggers local responses and activates sensory nerves to convey information to the central nervous system. However, little is known regarding participation of ATP and mediators of ATP release, such as Pannexin 1 (Panx1) channels, in mechanisms of vaginal mechanosensory transduction and of changes imposed by diabetes and menopause, conditions associated with vaginal dysfunction and risk for impaired genital arousal.AimTo investigate if intravaginal mechanical stimulation triggers vaginal ATP release and if (a) this response involves Panx1 channels and (b) this response is altered in animal models of diabetes and menopause.MethodsDiabetic Akita female mice were used as a type 1 diabetes (T1D) model and surgical castration (ovariectomy [OVX]) as a menopause model. Panx1-null mice were used to evaluate Panx1 participation in mechanosensitive vaginal ATP release. Vaginal washes were collected from anesthetized mice at baseline (non-stimulated) and at 5 minutes after intravaginal stimulation. For the OVX and Sham groups, samples were collected before surgery and at 4, 12, 22, 24, and 28 weeks after surgery. ATP levels in vaginal washes were measured using the luciferin-luciferase assay. Panx1 mRNA levels in vaginal epithelium were quantified by quantitative polymerase chain reaction.OutcomesThe main outcome measures are quantification of mechanosensitive vaginal ATP release and evaluation of impact of Panx1 deletion, OVX, and T1D on this response.ResultsIntravaginal mechanical stimulation–induced vaginal ATP release was 84% lower in Panx1-null (P < .001) and 76% lower in diabetic (P < .0001) mice compared with controls and was reduced in a progressive and significant manner in OVX mice when compared with Sham. Panx1 mRNA expression in vaginal epithelium was 44% lower in diabetics than that in controls (P < .05) and 40% lower in OVX than that in the Sham (P < .05) group.Clinical TranslationPanx1 downregulation and consequent attenuation of mechanosensitive vaginal responses may be implicated in mechanisms of female genital arousal disorder, thereby providing potential targets for novel therapies to manage this condition.Strengths & LimitationsUsing animal models, we demonstrated Panx1 involvement in mechanosensitive vaginal ATP release and effects of T1D and menopause on this response and on Panx1 expression. A limitation is that sex steroid hormone levels were not measured, precluding correlations and insights into mechanisms that may regulate Panx1 expression in the vaginal epithelium.ConclusionsPanx1 channel is a component of the vaginal epithelial mechanosensory transduction system that is essential for proper vaginal response to mechanical stimulation and is targeted in T1D and menopause.Harroche J, Urban-Maldonado M, Thi MM, et al. Mechanosensitive Vaginal Epithelial Adenosine Triphosphate Release and Pannexin 1 Channels in Healthy, in Type 1 Diabetic, and in Surgically Castrated Female Mice. J Sex Med 2020;17:870–880.     

雌激素、三苯氧胺与氟化物配伍预防去势大鼠骨丢失的研究

目的　比较三苯氧胺与雌激素对骨代谢的影响 ,及两者与氟化物配伍后是否产生协同效果。方法　将 142只 6月龄雌性SD大鼠行去势手术或假手术后随机分为 7组 (每组 19～ 2 1只 ) :(1)假手术组 ;(2 )去势组 ;(3)雌激素组 ;(4 )氟化物组 ;(5 )雌激素 +氟化物组 ;(6 )三苯氧胺组 ;(7)三苯氧胺 +氟化物组。治疗 12个月 ,行骨密度、腰椎骨组织形态计量学参数 (不脱钙骨切片 )、右股骨中段三点弯曲试验观察 ,并行子宫病理及血脂检查。结果　 (1)术后 12个月时 ,全身骨密度去势组为2 79mg/cm2 、治疗组为 2 86～ 2 98mg/cm2 ,腰椎骨密度分别为 2 32mg/cm2 、2 5 1～ 2 6 6mg/cm2 (P均<0 0 5 ) ;股骨中段骨密度 ,雌激素组 2 16mg/cm2 ,明显高于三苯氧胺组 195mg/cm2 (P <0 0 5 ) ;配伍治疗组与单药治疗组无明显差异 (P >0 0 5 )。 (2 )术后 4个月 ,两个配伍治疗组最大载荷 (均为 145牛顿 )与去势组 [(118± 2 4)牛顿 ]有显著差异 (P <0 0 5 ) ;术后 12个月各治疗组最大载荷为 132～ 15 5牛顿 ,均明显高于去势组 (10 8± 13)牛顿 (P <0 0 5 ) ,雌激素组最大载荷、弹性载荷均明显高于三苯氧胺组 (P <0 0 5 )。 (3)各组骨组织形态计量学检查未发现骨矿化不良现象。结论　雌激素在维持骨量、骨强度方面优于三苯     

Effects of strontium ranelate,raloxifene and misoprostol on bone mineral density in ovariectomized rats

Objectives

To investigate the effects of strontium ranelate, raloxifene and misoprostol on bone mineral density (BMD) in ovariectomized rats to contribute to the individualization of the treatment of postmenopausal osteoporosis.

Study design

Sixty sexually mature female Sprague–Dawley rats weighing 250 g were used. The 60 rats were divided into six groups of 10 rats each: SR, MISO, RAL, SHAM, DW and OVX. All except the SHAM rats were subjected to bilateral ovariectomy. Three days after surgery, rats were administered strontium ranelate (Protelos^®, 2 g, Servier, Istanbul), 1800 mg/kg/day; misoprostol (Cytotec^®, 200 mcg, Ali Raif, Istanbul), 200 mcg/kg/day; raloxifene (Evista^®, 60 mg, Lily and Company, Istanbul), 3 mg/kg/day and 1 cc of distilled water by gavage for 8 weeks. Bone mineral density measurements were then performed.

Results

The strontium ranelate (SR) group had significantly higher vertebral BMD than all other groups. Femoral density in the SR group was also significantly higher than in other groups and there was no difference between femoral density in the strontium ranelate and sham groups.

Conclusions

Strontium ranelate, raloxifene and misoprostol can prevent bone loss in the vertebrae, whereas strontium ranelate can also prevent bone loss in the femur of ovariectomized rats. Strontium ranelate increases greater than raloxifene and misoprostol BMD in the vertebrae.

Condensation

Strontium ranelate may increase both vertebral and femur BMD in ovariectomized rats while raloxifene and misoprostol may only increase lumbar spine BMD.     

Protective effects of beta glucan in brain tissues of post-menopausal rats: a histochemical and ultra-structural study

Decline of estrogen during menopause has been associated with numerous significant changes that have been linked to many pathophysiological complications. In addition, ovarian hormone deficiency increases the production of reactive oxygen radicals which could result in oxidative stress and cell damage. While estrogen therapy is often considered to overcome the behavioral and physiological shortcomings, antioxidants are gaining popularity for their beneficial property. For this purpose, in the present study, utilizing the antioxidant properties of beta glucan has been examined in treatment of menopause induced oxidative stress in cerebral neurons. Four groups of female Wistar rats were used: control, ovariectomy, ovariectomy?+?estrogen treated and ovariectomy?+?beta glucan treated. We observed a significant increase in neural degeneration in ovariectomized rats as compared to controls. Moreover, increased oxidative stress in the brains of the ovariectomized rats has been detected by performing immunohistochemical analysis. A large number of immuno-positive cerebral neurons have been observed in ovariectomy group rat brains. Interestingly, providing beta glucan treatment to ovariectomized rats reduced the number of degenerated neurons. Our study is the first to examine light and electron microscopic examination and immunohistochemical and stereological analysis of estrogen depletion in rats and to test protective role of beta glucan in the experimental study.     

Ultra-structural changes and apoptotic activity in cerebellum of post-menopausal-diabetic rats: a histochemical and ultra-structural study

Diabetes mellitus (DM) is one of the most common and chronic diseases, especially in post-menopausal periods. Neuro-degeneration occurs more frequently in post-menopausal diabetics. Therefore, we investigated ovariectomized rats cerebellar cortex response to the estradiol deficiency and hyperglycemia. For the ovariectomy, the rats were bilaterally ovariectomized, and then DM induced by a single dose of Alloxan monohydrate injection in ovariectomy or/and diabetic groups. During light and electron microscopic examination, degenerated Purkinje cells membrane, swollen organelles, degenerated mitochondria, edema formation and vacuolization were seen in the ovariectomy and ovariectomy-diabetic groups sections. In addition, increased apoptotic activity was observed in the ovariectomy and ovariectomy-diabetic groups compared to the control group. We demonstrated that estradiol and insulin deficiency can affect the cerebellar cortex, which support the hypothesis that the execution of neuronal damages in post-menopausal diabetics. Also, diabetes and menopause are major risks factors for many disorders including nervous system and the number of post-menopausal-diabetics are increasing world-wide.     

替勃龙对卵巢切除大鼠膀胱功能及结构的影响

目的:通过比较正常、卵巢切除及卵巢切除后补充替勃龙大鼠的膀胱功能和组织形态,探讨替勃龙对膀胱的影响及其作用机制。方法:30只雌性成年SD大鼠分为3组:正常对照组、切除双侧卵巢组(OVX组)、切除双侧卵巢后补充替勃龙组(OVX+Tib组)。用药12周后测定膀胱压力容积,Masson染色膀胱石蜡切片分析胶原纤维(CF)和平滑肌(SM)的面密度及二者比值。结果:(1)OVX组膀胱最大容量(0.32±0.20m l)、顺应性(0.012±0.006m l/cmH2O)和最大收缩力(1.4±0.4cmH2O)相对于正常对照组(分别为1.11±0.09m l、0.026±0.003m l/cmH2O和4.4±0.3cmH2O)明显减少,有统计学差异(P<0.01)。OVX+Tib组膀胱顺应性(0.022±0.003m l/cmH2O)与正常对照组比较无统计学差异(P>0.05),膀胱最大容量(0.87±0.26m l)及最大收缩力(3.3±1.0cmH2O)比正常对照组减少(P<0.05),但比OVX组增加(P<0.01);(2)CF面密度、CF面密度与SM面密度比值:OVX组(0.2180±0.0407和0.5396±0.0837)比正常对照组(0.1598±0.0387、0.3199±0.0860)增加,有统计学差异(P<0.01)。OVX+Tib组此两值(0.1893±0.0251、0.4249±0.0646)介于OVX组与正常对照组之间。结论:大鼠切除双侧卵巢后膀胱功能降低,补充替勃龙在一定程度上改善了膀胱功能,这可能是与它对胶原的抑制有关。     

Exogenous testosterone and estrogen affect bladder tissue contractility and histomorphology differently in rat ovariectomy model

IntroductionChanges in sex hormone levels may play a role in the etiology of lower urinary tract dysfunction of aging women where the possible role of testosterone is overlooked.AimTo determine the effect of testosterone with/without estrogen replacement on histological and functional deterioration in ovariectomized rat bladder tissue.MethodsA total of 54 female Sprague Dawley rats were divided into 6 groups. Except sham operated (control group), all others underwent bilateral ovariectomy. No further treatment was given to the ovariectomy-only group (OVX group). At the third week of ovariectomy treatments were started; vehicle agent (VA group), estradiol (E2 group), testosterone undecanoate (T group), and estradiol + testosterone undecanoate combination (E2 + T group) in physiological doses. Nine weeks after ovariectomy, bladder strips were harvested for isometric tension and histopathological studies.Main Outcome MeasuresTo assess the effect of testosterone/estradiol on ovariectomized rat bladder tissue function and histomorphology.ResultsOVX and VA groups showed statistically significant histological changes such as urothelial damage, inflammatory cell infiltration, increase in collagen fibers and muscular atrophy compared with the control group. Both E2 and T reversed these changes but best histomorphological restoration was observed in E2 + T group. In isometric tension studies, ovariectomy tended to increase contractile responses which were normalized after E2 treatment. In contrary to E2, T significantly increased contractile responses that were normalized with combination treatment. During relaxation studies statistically significant higher relaxation responses were observed in ovariectomized rats. Although both exogenous testosterone and estradiol tended to reverse this effect, a statistically significant difference was found only after testosterone treatment.ConclusionEither estradiol or testosterone replacement alone or in combination prevents significant alterations in bladder tissue histology following ovariectomy whereas both affect the bladder tissue contractility. Thus, combination treatment appears to be the best method to restore both contractility and histomorphology of bladder tissue after ovariectomy. Tanidir Y, Ercan F, and Tarcan T. Exogenous testosterone and estrogen affect bladder tissue contractility and histomorphology differently in rat ovariectomy model.     

Effects of Ovariectomy and Dehydroepiandrosterone (DHEA) on Vaginal Wall Thickness and Innervation

IntroductionOne mechanism by which low sexual steroid activity observed after menopause could cause sexual dysfunction is by deficient vaginal innervation. Recently, it has been shown that intravaginal administration of dehydroepiandrosterone (DHEA) could produce beneficial effects on sexual dysfunction in postmenopausal women.AimThe goal of this study was to determine if DHEA could modify innervation in the rat vagina.Main Outcome MeasuresThe area occupied by the nerve fibers immunoreactive for protein gene product 9.5 (PGP 9.5), a panneuronal marker or tyrosine hydroxylase (TH), a sympathetic nerve fiber marker, in the lamina propria and muscular layers, respectively, as well as the total area of each of these 2 layers were measured by stereological analysis.MethodsThe innervation of the rat vagina was examined 9 months after ovariectomy (OVX) compared to intact animals and treatment of OVX animals with DHEA (80 mg/kg). Four sections from each vagina (5 animals/groups) were immunostained.ResultsIn OVX animals, the lamina propria area was decreased to 44%, an effect which was reversed by DHEA to 69% of the intact value. OVX also caused a 59% decrease in the area of PGP 9.5 fibers, an effect which was prevented by DHEA, thus showing a 68% stimulatory effect of DHEA on the density of PGP 9.5 fibers in the lamina propria compared to OVX animals. Following OVX, the muscular layer area was decreased by 61%. DHEA treatment induced 118% and 71% increases in TH fiber area compared to OVX and intact animals, respectively. The density of TH fibers was 182% increased over intact controls by DHEA treatment of OVX animals.ConclusionsThe relatively potent stimulatory effect of DHEA on intravaginal nerve fiber density provides a possible explanation for the beneficial effects of intravaginal DHEA on sexual dysfunction observed in postmenopausal women. Pelletier G, Ouellet J, Martel C, and Labrie F. Effects of ovariectomy and dehydroepiandrosterone (DHEA) on vaginal wall thickness and innervation.     

雷洛昔芬对去卵巢大鼠骨密度及血E2、IL-6水平影响的研究

目的观察雷洛昔芬对去势大鼠骨密度、血清E2、IL-6及子宫内膜的影响，探讨雷洛昔芬抗骨质疏松的作用机制。方法48只3个月龄雌性SD大鼠，随机分为SHAM组（假手术），0VX组（单纯去势），0VX＋E2组（去势＋雌激素），0VX＋R组（去势＋雷洛昔芬），每组12只。3个月后处死采集血，用放射免疫分析法检测并比较各组E2及IL-6的表达，取子宫内膜用HE染色法观察各组子宫内膜的形态变化并对子宫内膜腺体数目进行统计学分析。结果雷洛昔芬组大鼠骨密度显著高于去卵巢组（P＜0．05），血清IL-6浓度值显著降低，雌激素浓度的改变差异无统计学意义（与去卵巢组比较P＞0．05），对子宫内膜无明显影响（P＞0．05）。结论雷洛昔芬治疗骨质疏松的作用与血清中IL-6水平降低相关，与雌激素无显著相关，对子宫内膜无显著影响。     

Endothelium-dependent and independent enhancement of vascular contractility in the ovariectomized rabbit

OBJECTIVE: Estrogen suppresses contractile response and increases vasodilator response, partly by modulating endothelial function. However, the effect of estrogen on the contractility of vascular smooth muscle remains to be elucidated. We investigated the effect of a long-term estrogen deficiency on vascular contractility and the Ca(2+) sensitivity of the contractile apparatus in arterial smooth muscle. METHODS: Female rabbits were divided into the following three groups: control group, an ovariectomized group (OVX), and a group supplemented with 17beta-estradiol after ovariectomy (OVX+E2). Twelve weeks later, the mesenteric artery was isolated, and the vascular contractility was evaluated. RESULTS: In OVX, the contractile responses to phenylephrine and 118 mM potassium were enhanced, and the basal release of nitric oxide decreased in the strips with endothelium compared with either OVX+E2 or control. An enhancement of contraction was also observed in the strips without endothelium. However, the extent of enhancement was smaller than that observed in the presence of endothelium. The simultaneous measurement of calcium ([Ca(2+)](i)) and tension revealed no significant difference in the [Ca(2+)](i) elevations induced by phenylephrine among the three groups. In the alpha-toxin permeabilized strips, the Ca(2+)-tension relationships obtained both with and without phenylephrine and guanosine triphosphate were similar among the three groups. No difference in the myosin expression and the histology of vascular tissue was observed among the three groups. CONCLUSION: Long-term estrogen deficiency increased the vascular tone mainly by enhancing smooth muscle contractility. Endothelial dysfunction is considered to play a minor role in the augmentation of vascular tone.     

胰岛素抵抗及子宫内膜增殖大鼠模型中脂肪因子和PI3K/AKT信号通路相关蛋白的表达

目的:观察有胰岛素抵抗及子宫内膜增殖大鼠模型中血浆脂联素(Adipo)等脂肪因子的水平,检测Adipo和脂联素受体(AdipoR)在大鼠子宫组织中的表达,并探讨其分子生物学机制。方法:选取8周龄Sprague-Dawley(SD)大鼠45只(每组9只),分别给予普通饮食(StD组)和高脂饮食(HFD组)40周。将StD组分为对照C组(普通饮食+假去势手术+对照溶剂)、NO组(普通饮食+去势手术+对照溶剂)、NE组(普通饮食+去势手术+17β-雌二醇灌胃),HFD组分为FO组(高脂饮食+去势手术+对照溶剂)和FE组(高脂饮食+去势手术+17β-雌二醇灌胃)。应用酶联免疫吸附(ELISA)检测小鼠血浆中脂肪因子水平,实时荧光定量PCR、Western Blotting和免疫组化检测子宫内膜中脂肪因子的水平,免疫组化检测子宫内膜中PTEN、AMPK和PI3K/AKT信号通路相关蛋白的表达。结果:①与NE组大鼠相比,FE组大鼠子宫腔上皮细胞和腺上皮细胞的高度、肌层的厚度显著增加(P<0.05)。②与对照C组相比,子宫内膜增生组(NE组和FE组)大鼠血浆Adipo水平显著降低(P<0.05),FO组大鼠血浆Adipo显著升高(P<0.05)。③在雌激素的作用下(NE组和FE组)子宫组织中Adipo mRNA的表达增多:与NO组大鼠相比,NE组大鼠子宫组织中Adipo mRNA水平显著升高(P<0.05);与FO组大鼠相比,FE组大鼠Adipo mRNA水平也显著升高(P<0.05)。大鼠子宫中AdipoR1 mRNA和AdipoR2 mRNA水平在各组之间差异无统计学意义(P>0.05)。与对照C组相比,FE组大鼠子宫组织中Adipo蛋白的表达明显降低(P<0.05)。④FE组子宫PTEN和p-AMPK蛋白的表达显著低于对照C组(P<0.05),PI3Kp85α、p-AKT蛋白表达显著高于对照C组(P<0.05)。结论:长期高脂饮食诱导大鼠的胰岛素抵抗可协同17β-雌二醇刺激子宫内膜增殖;胰岛素抵抗和雌激素影响血浆Adipo的水平;胰岛素抵抗和雌激素的协同作用使大鼠子宫内膜PTEN的表达降低、AMPK通路可能被抑制、PI3K/AKT信号通路可能被激活。     

卵巢切除和雌激素补充治疗对大鼠膀胱功能及结构的影响

目的通过对不同雌激素水平大鼠进行膀胱功能、组织形态和超微结构的比较,探讨雌激素对膀胱功能的影响及其作用机制。方法30只雌性成年SD大鼠均分为3组:OVX+E组(切除双侧卵巢后补充戊酸雌二醇0·8mg·kg-1·d-1,溶于0·5%羧甲基纤维素钠,每日灌胃1次)、OVX组(切除双侧卵巢)、正常对照组(未切除卵巢),后两组大鼠每日给予0·5%羧甲基纤维素钠灌胃1次。3组大鼠用药12周后行膀胱压力容积测定,并用切除膀胱的石蜡切片分析胶原纤维和平滑肌的面密度及两者比值,透射电镜下观察逼尿肌的超微结构。结果(1)OVX组膀胱最大容量(0·32±0·20)ml、顺应性(0·012±0·006)ml/cmH2O(1cmH2O=0·098kPa)和最大收缩力(1·4±0·4)cmH2O,相对于正常对照组[分别为(1·11±0·09)ml、(0·026±0·003)ml/cmH2O和(4·4±0·3)cmH2O]明显减少,差异均有统计学意义(P<0·05)。OVX+E组膀胱最大容量为(0·83±0·10)ml,相对于正常对照组减少(P<0·05),而膀胱顺应性(0·029±0·003)ml/cmH2O、最大收缩力(4·8±1·4)cmH2O与正常对照组比较,差异无统计学意义(P>0·05)。(2)胶原纤维面密度、胶原纤维面密度/平滑肌面密度比值,OVX组分别为0·218±0·041和0·54±0·08,相对于正常对照组(0·160±0·039、0·32±0·09)明显增加,差异有统计学意义(P<0·05);而OVX+E组(0·178±0·027、0·38±0·06)与正常对照组比较,差异无统计学意义(P>0·05)。(3)电镜观察OVX组逼尿肌超微结构出现退行性改变,其他两组无类似变化。结论大鼠切除双侧卵巢后膀胱功能明显降低,补充雌激素有利于改善膀胱功能,这一作用可能是通过抑制胶原增生,保护细胞器来实现的。     

雌激素对去卵巢大鼠骨丢失的骨形态计量学及组织学的影响

OBJECTIVES: To demonstrate the histomorphometric and histological changes of bone 3 weeks after bilateral ovariectomy in rats and to investigate the impacts of 4 different hormone replacement therapies on the bone histomorphometric, histological appearances. METHODS: Bilateral ovariectomies were done on 41 female rats and sham operations on other 9 (sham group) respectively. After 3 weeks, 4 different treatments: i.e. Livial, Gevrine, Premarin, Weinian were initiated separately on each 8 ovariectomized rats for another 3 weeks. The remaining 9 were served as controls (OVX group). All rats were sacrificed either 3 weeks after ovariectomy/sham operation or at the end of hormone therapies. Their femoral bone mineral density (BMD) were measured by dual energy X-ray absorptiometry (DEXA). Specimens of proximal femur were embedded undecacifide for histomorphometric analysis and of distal femoral metaphysics were procured for scanning electron microscope (SEM) and pathologic examinations. RESULTS: (1) Three weeks after OVX, the femoral BMD, mean cortical thickness decreased significantly while the number of osteoclast increased significantly as compared with sham group. The trabecular became thinner and irregular which changed the bone microstructure in three dimension. (2) After treatment of 4 different preparations, the above parameters restored to various extents to the sham operation levels. Among them, there was greater increase of femoral BMD on the Livial and Gevrine group as compared with Premarin and Weinian group (P < 0.05). CONCLUSIONS: Bilateral ovariectomy induced increased osteoclast activity and bone turnover, therefore caused accelerated bone loss. Treatment with combined sex hormones preparation could inhibit bone absorption and stimulate bone formation, especially those containing androgenic activity could increase the BMD.     

Effect of immune stress on body weight regulation is altered by ovariectomy in female rats

It has been suggested that obesity and loss of ovarian function alter the inflammatory response to immune stress. Ovariectomized (OVX) rats, which are used as a model of human menopause, exhibit both hyperphagia-induced obesity and gonadal steroid deficiency. To evaluate the effects of ovariectomy on inflammatory responses, we compared the anorectic response to LPS in OVX rats and gonad intact female rats. As leptin and hypothalamic interleukin-1β (IL1β) play pivotal roles in the anorectic response to immune stress, these factors were also measured. It was found that the OVX rats exhibited an increased anorectic response to LPS compared with the sham-operated rats. The OVX rats showed higher serum leptin concentrations and a greater increase in hypothalamic IL1β mRNA expression after LPS injection. In addition, in order to determine whether gonadal steroid deficiency contributes to the changes in the inflammatory responses of OVX rats, we compared responses between OVX rats treated with gonadal steroids and untreated OVX rats. There were no differences in appetite, the serum leptin level, and hypothalamic IL1β mRNA expression between the two groups after LPS injection. These findings suggest that the loss of ovarian function increases the induction of leptin and hypothalamic IL1β synthesis and consequently increases the anorectic response under immune stress conditions. It is possible that these alterations are caused by OVX-induced obesity rather than the direct effects of gonadal steroid deficiency.     

戊酸雌二醇对去卵巢大鼠骨质疏松的治疗作用

目的评价国产戊酸雌二醇对去卵巢大鼠骨质疏松的治疗作用.方法雌性健康未交配Wistar大鼠42只,于6月龄时分别行去卵巢和假手术,分组并行如下处理(1)治疗前对照组(8只)于去卵巢后6周处死;(2)治疗末对照组(8只);(3)假手术组(8只);(4)去卵巢17β雌二醇治疗(O+E)组(9只)去卵巢后6周予17β雌二醇20μg@kg-1@d-1皮下注射;(5)去卵巢戊酸雌二醇治疗(O+EV)组(9只)去卵巢后6周予戊酸雌二醇800μg/kg@d-1经胃肠道给药,后4组于去卵巢后14周处死,留取右侧胫骨、股骨及血标本,处死前收集24h尿标本,分别进行骨组织计量学、外周骨定量CT检查、股骨远端凹入实验,以及骨吸收指标脱氧吡啶啉等测定.结果去卵巢后,骨矿盐含量(骨量)减少,骨小梁结构稀疏,骨吸收和骨形成指标均增加.股骨远端松质骨骨量和骨矿盐密度(骨密度)显著降低,松质骨最大载荷和刚度均明显下降.与治疗末对照组相比,O+E和O+EV组骨吸收指标尿脱氧吡啶啉分别下降54.6%和77.4%(P均＜0.01),骨转换受抑制,胫骨近端次级骨小梁的体积比率分别增加122.3%和119.7%[(12.9±0.8)%和(12.8±0.9)%与(5.8±1.3)%比较,P＜0.01],骨小梁的结构得到改善.O+E和O+EV组股骨远端松质骨骨矿盐密度较治疗末对照组分别增加99.5%和128.4%,分别为(258.9±16.9)和(269.2土24.3)mg/cm2比(117.8±30.4)mg/cm3(P＜0.01).O+E组的股骨远端松质骨最大载荷和刚度均较治疗末对照组有显著增加(P均＜0.05),以上各指标在0+E和0+EV组之间差异无显著性.结论戊酸雌二醇能有效地抑制去卵巢大鼠过高的骨转换,使小梁骨骨量增加,结构改善,松质骨的力学特征得到改善.