不同培养方法骨髓破骨细胞样细胞分化及活性的实验观察

目的研究不同骨髓细胞培养方法对大鼠破骨细胞样细胞(OLC)诱导分化及活性的影响。方法大鼠骨髓细胞以巨噬细胞集落刺激因子(M-CSF)作用后与核因子!B受体活化因子配体(RANKL)协同诱导OLC分化,采用全骨髓细胞(A组)及密度梯度离心后骨髓单个核细胞(B组)两种培养方法,通过倒置相差显微镜及细胞染色观察两组OLC分化数量的差异,通过骨吸收陷窝分析及OLC膜表面整合素#3(CD61)表达量比较两组OLC活性的差异。结果B组培养5、7d抗酒石酸酸性磷酸酶(TRAP)染色阳性多核OLC数量及TRAP活性均高于A组,骨吸收陷窝计数亦较A组为多,差异均有统计学意义(P<0.01,P<0.05);陷窝面积比培养早期两组比较差异无统计学意义,7d时B组大于A组,差异有统计学意义(P<0.01);OLC膜表面CD61表达量及平均荧光强度0及3d时B组均较A组明显增高,随培养时间延长组间比较差异无统计学意义。结论低转速、短时间密度梯度离心后骨髓细胞培养诱导OLC分化获得的细胞数量较多且不影响其活性,是一种简便有效的OLC培养方法。     

雌激素对大鼠去卵巢后白细胞介素-6介导的骨吸收的影响

目的观察雌激素对去卵巢大鼠骨髓细胞白细胞介素6(IL6)、IL6受体、gp130基因表达以及骨髓源性破骨细胞形成的影响。方法健康3月龄雌性SD大鼠72只,随机平均分为假手术对照组、去卵巢组和雌激素组(苯甲酸雌二醇02mg/kg,皮下注射,每周1次)。分别于术后2、4、6、12周每组各取6只大鼠骨髓细胞作细胞培养和提取RNA。培养第6天计数破骨细胞数,第12周取左侧胫骨作骨形态计量学检测。结果骨形态学显示去卵巢后出现骨吸收亢进,雌激素对其有抑制作用。去卵巢后2周,去卵巢组破骨细胞形成数即多于对照组(1450±169对901±141,P<005),骨髓细胞IL6和IL6受体mRNA表达均显著升高(P<005,P<001),第4～6周,上述改变达高峰,至第12周仍呈有意义增高;从2～12周,上述各指标雌激素组均明显低于去卵巢组(P<005,P<001)。各组未见gp130基因表达水平有明显变化。结论雌激素可以抑制大鼠去卵巢后骨髓源性破骨细胞的生成,这一效应可能与其抑制骨髓细胞在去卵巢后过度表达IL6、IL6受体基因有关。     

两种大鼠骨髓源破骨样细胞诱导方法的比较

目的 比较两种不同的大鼠骨髓源破骨样细胞(Osteoclast-like cells,OLC)体外分离培养的方法.方法 收集5周龄SD大鼠骨髓细胞悬液,加入M-CSF(10 ng/ml)培养24 h后置入不同的诱导培养体系中,即A组:巨噬细胞集落刺激因子(M-CSF)+破骨细胞分化因子(RANKL),B组:M-CSF+1,25二羟基维生素D3[1,25(OH)2D3]+地塞米松(Dexamethasone),分别于培养3、5、7、9、12 d利用抗酒石酸碱性磷酸酶(TRAP)染色、骨吸收陷窝检测等对获得的OLC进行形态学和功能观察,并进行计数比较.结果 两种方法 均可诱导出TRAP染色阳性的多核细胞.两组细胞数量均于第7天达到最高峰,B组于7、9、12 d获得的细胞数量较A组多(P<0.05);B组骨吸收陷窝数于9、12 d时多于A组(P<0.05).结论 两种方法 均诱导出破骨样细胞,B组诱导OLC的细胞活性和细胞数量高于A组.     

去卵巢大鼠骨质疏松凋亡细胞及其相关因素观察

目的　探讨去卵巢大鼠骨细胞凋亡活性及其相关因素。方法　采用3′-OH末端DNA原位标记技术观察凋亡细胞活性;采用免疫组化SABC法观察bcl-2、Fas、转化生长因子（TGF）β1的表达情况。结果　去卵巢大鼠成骨细胞的凋亡细胞活性为（40.5±5.2）%,较假手术组(24.5±3.0)%与去卵巢+尼尔雌醇/左炔诺孕酮治疗组［OVX+N/L组,（26.4±2.9)%］明显增加,差异有显著性(P＜0.01);去卵巢大鼠破骨细胞的凋亡细胞活性为（8.4±1.2）%,明显低于假手术组(24.0±2.9)%与OVX+N/L组(26.5±3.1)%,差异具有显著性(P＜0.01)。3组成骨细胞与破骨细胞内bcl-2阳性表达率均较低,差异无显著性(P值均>0.05);去卵巢大鼠组成骨细胞内Fas(80.0%)高于假手术组（40.0%）和OVX+N/L组（40.0%）,而破骨细胞内Fas(20.0%)低于假手术组(70.0%)和OVX+N/L组(73.3%),后者差异有显著性(χ2=7.94,P＜0.05),OVX组成骨和破骨细胞内TGFβ1(分别为20.0%、0),均低于其他两组,前者差异有极显著意义(χ2=13.104,P＜0.01)。结论　去卵巢大鼠骨丢失主要是由于雌激素水平低下引起破骨细胞凋亡减少、成骨细胞凋亡活性增加致骨吸收功能明显增加而超过骨形成所致;TGFβ1的分泌可能需要雌激素的刺激,TGFβ1表达可能抑制成骨细胞凋亡,促进破骨细胞凋亡,Fas可能诱导破骨细胞凋亡。     

雌激素对兔动脉粥样硬化形成及某些体液因子的影响

目的　建立雌激素抗兔动脉粥样硬化模型 ,阐明雌激素作用的可能机制。　方法　 2 1只纯种新西兰兔随机分入 :A组 (切除卵巢未补充雌激素 )、B组 (切除卵巢补充雌激素 )、C组 (未切除卵巢 ) ,给予 1%胆固醇饮食 ,8周时比较动脉形态学变化 ,测定动脉一氧化氮及一些体液因子。　结果　 (1) A组兔主动脉内膜面积〔(1.35± 0 .44 ) m m2 〕高于 B、C组〔(0 .2 0± 0 .2 9)及 (0 .34± 0 .2 9) mm2 〕(P<0 .0 5 ) ;(2 ) A组兔内皮素 - 1高于 B组及 C组 (P<0 .0 5及 P<0 .0 1) ,血浆肾素活性、血管紧张素 高于 C组 (P<0 .0 5 ) ;(3) B、C组血浆雌二醇〔(2 12± 78)及 (2 2 9± 10 3) ng/ L〕和主动脉一氧化氮〔(0 .41± 0 .18)及 (0 .44± 0 .16 ) nmol/ m g〕高于 A组〔(10 3± 6 9) ng/ L及 (0 .2 5± 0 .0 6 ) nm ol/ m g〕(P<0 .0 5 )。　结论　雌激素可明显抑制动脉粥样硬化形成 ,可能与促进一氧化氮生成、抑制内皮素生成、抑制肾素血管紧张素系统活性有关。     

激素替代治疗对去卵巢大鼠动脉雌激素受体表达的影响

目的　研究雌、孕激素替代治疗时去卵巢大鼠动脉壁雌激素受体 (ER)的表达。　方法　将 40只成年雌性大鼠随机分为 4组 :假手术对照组、去卵巢组、去卵巢 雌激素治疗组、去卵巢 雌孕激素治疗组。给予正常饮食 ,2个月后处死大鼠。用放免法检测血清中雌二醇和孕酮浓度 ,用放射配体结合分析法检测大鼠主动脉ER的含量。　结果　 (1)正常雌性成年大鼠 (假手术对照组 )主动脉中存在ER ,解离常数 (Kd)为 (3 0± 0 7)× 10 -9,最大结合容量 (Bmax)为 (37 4± 7 9)pmol/g蛋白 ;(2 )去卵巢组大鼠动脉ER的Bmax〔(10 4± 5 9)pmol/g蛋白〕显著低于假手术组 (P <0 0 1) ;(3)去卵巢 雌激素治疗组与去卵巢 雌孕激素治疗组动脉ER的Bmax差别无显著性〔(分别为 (40 7± 7 5 )和(35 1± 4 5 )pmol/g蛋白〕 ,二者都显著高于去卵巢组 ;各组间Kd值差别无显著性。　结论　去卵巢2个月后大鼠主动脉的ER表达显著减少 ,补充雌激素或补充雌、孕激素能使去卵巢大鼠主动脉ER的含量维持在正常水平 ,这可能是雌激素对绝经后妇女心血管保护作用的机制之一。     

补肾中药对去卵巢大鼠骨髓IL-6、IL-6R基因表达及骨髓源性破骨细胞形成的影响

目的　观察补肾中药对去卵巢大鼠骨髓细胞表达白介素 6 (IL 6 )、IL 6受体 (IL 6R)、gp130基因的影响及其与骨髓干细胞诱导分化形成破骨细胞能力变化的关系。　 方法　健康 3月龄雌性SD大鼠 ,随机分为假手术对照组、去卵巢组、雌激素组、中药组。取骨髓细胞作细胞培养和提取RNA。细胞培养第 6天染色 ,以TRAP染色 (+)以及胞核≥ 3为破骨细胞。骨髓细胞用TRIZOL提取总RNA后进行RT PCR。　结果　去卵巢组术后 6周及 12周 ,骨髓破骨细胞形成数明显多于对照组 (P <0 0 5 ) ,补肾中药与雌激素均能抑制破骨细胞的形成 (P <0 0 5 )。去卵巢后 6周骨髓细胞IL 6和IL 6RmRNA表达显著增高 (P <0 0 5及 0 0 1) ,第 12周时 ,IL 6、IL 6R基因表达虽有下降 ,但仍然高于对照组。补肾中药与雌激素均显示可减少上述基因过度表达 ,以第 6周时作用最显著(P <0 0 5及 0 0 1)。以上各组全程未见 gp130基因表达水平有明显变化。　 结论　本方所用补肾药物可以抑制大鼠去卵巢后骨髓IL 6、IL 6R基因过度表达 ,这一效应与药物抑制骨髓源性破骨细胞生成的作用密切相关。     

吡格列酮对体外培养大鼠破骨细胞样细胞分化及活性的影响

目的 观察吡格列酮对大鼠破骨细胞样细胞(OLC)分化及活性的影响,探讨PPARγ2活化与破骨细胞之间的关系.方法 用NF-кB受体活化因子配体(RANKL)及巨噬细胞集落刺激因子(M-CSF)协同诱导大鼠骨髓单个核细胞(BMC)向OLC分化,同时加入1、5、10μmol/L盐酸吡格列酮干预.应用RT-PCR分析OLC分化过程中PPARγ2及NF-кB受体活化因子(RANK)mRNA的表达,结合细胞染色及抗酒石酸酸性磷酸酶(TRAP)活性观察吡格列酮对OLC分化的影响,通过骨吸收陷窝分析及整合素β3(CD61)平均荧光强度的检测比较吡格列酮对OLC活性的影响.结果 (1)对OLC分化的影响:培养7 d时10μmol/L吡格列酮组与其它组相比OLC数量及TRAP活性明显减少(P<0.01或P<0.05),表明吡格列酮部分抑制OLC分化且此作用与剂量相关;RT-PCR结果显示在诱导分化早期,不同浓度吡格列酮呈剂量依赖性上调PPARγ2的表达水平,但随OLC的成熟其表达渐减弱,而RANK表达在5及10 p,mol/L吡格列酮干预组培养的各时间点均明显下调,与对照及1μmol/L干预组相比有显著差异(P<0.05或P<0.01);(2)对OLC活性的影响:骨吸收陷窝数量及吸收面积在吡格列酮5、10μmol/L组明显减少,与另两组相比差异显著(P<0.05或P<0.01),培养早期这两组细胞CD61平均荧光强度均明显下调(P<0.05或P<0.01).结论 吡格列酮激活PPAR田γ2转录活性可部分抑制大鼠骨髓OLC的分化及活性.     

雌激素对体外培养破骨细胞功能的影响

目的　探讨雌激素对体外培养破骨细胞功能的影响。　方法　利用酶动力学法测定培养基中酸性磷酸酶 (ACP)和抗酒石酸酸性磷酸酶 (TRAP)活性 ;共聚焦显微镜观察破骨细胞内氢离子随雌激素浓度的变化情况 ;LeicaQuantimet 5 0 0图像分析系统对骨吸收陷窝进行图像分析 ;扫描电镜观察骨吸收陷窝变化。　结果　随着雌激素浓度的升高 ,处理组与对照组ACP和TRAP的活性分别从(1 6 9± 0 13)U/L和 (1 6 0± 0 14)U/L降低到 (1 16± 0 31)U/L和 (0 93± 0 34)U/L(均为P <0 0 1) ,骨吸收陷窝的数目与面积从 (13 2 5± 1 5 2 )个 /片和 (4 82 2 7± 2 2 5 2 3) μm2 减少到 (4 6 8± 0 2 4)个 /片与(35 6 35± 145 41) μm2 ,处理组与对照组比较差异具有显著性意义 (P <0 0 1)。破骨细胞相对荧光值经过短暂的下降后持续上升 ,在第 12 0 0s时达到最高值。　结论　雌激素能够抑制氢离子释放 ,同时能改变ACP和TRAP活性 ,因而能直接抑制破骨细胞的骨吸收功能。     

Light诱导类风湿关节炎滑膜细胞向破骨细胞转化

目的 观察Light在类风湿关节炎(RA)滑膜细胞向破骨细胞转化过程中的作用.方法 取8例RA患者滑膜组织,胶原酶消化获取滑膜细胞,每例滑膜细胞分成5份培养,第1组加入巨噬细胞集落刺激因子(MCSF)作阴性对照,第2组加入MCSF和LIGHT,第3组加入MCSF和核因子(NF)-κB受体激动剂配体(RANKL),第4组加入MCSF、LIGHT和RANKL,第5组加入LIGHT.体外培养2周后,行抗酒石酸酸性磷酸酶(TRAP)染色,F肌动蛋白(F-actin)染色以及象牙片上骨吸收陷窝观察破骨细胞的形成和活性.结果 第1组和第5组TRAP(-),F-actin(-),象牙片上无骨吸收陷窝形成;第2组TRAP(+),F-actin(+),骨陷窝形成(+),多核破骨细胞呈圆形和椭圆形,体积较小,骨陷窝分散,体积较小;第3组TRAP(++).F-actin(++),骨陷窝形成(++),多核破骨细胞体积大,骨陷窝较多,体积大,形态不规则;第4组TRAP(+++),F-actin(+++),骨陷窝形成(++++),多核破骨细胞更多,骨陷窝大且有融合趋势.结论 Light能诱导RA滑膜细胞向破骨细胞转化,并能促进RANKL诱导滑膜细胞向破骨细胞转化的能力.     

Effects of chronic prednisolone treatment on bone resorption and bone composition in intact and ovariectomized rats and in ovariectomized rats receiving beta-estradiol

To examine the interactions between estrogen deficiency and glucocorticoid excess on bone metabolism the osteopenic effects of a standard dose of prednisolone (2 mg/kg BW.day) were studied in sham-ovariectomized (Sham-OVX), ovariectomized (OVX), and OVX rats given replacement beta-estradiol (OVX + E2). For 12 weeks six groups of female albino rats aged 4 months which had their skeletons labeled with 45Ca were fed matched amounts of low-calcium (0.1% Ca) hydroxyproline-free diet. The six treatment groups were: group 1, Sham-OVX; group 2, Sham-OVX + prednisolone; group 3, OVX; group 4, OVX + prednisolone; group 5, OVX + E2; group 6, OVX + E2 + prednisolone. Bone resorption was estimated by studying the urinary excretion of hydroxyproline and 45Ca. Parathyroid function was assessed indirectly from urinary cAMP excretion. Treatments did not influence parathyroid activity or serum levels of calcium or 1,25-dihydroxyvitamin D. However, ovariectomy increased bone resorption and induced osteopenia whereas prednisolone decreased bone resorption and formation and caused osteopenia. Ovariectomy increased the rate of bone resorption in prednisolone-treated rats; prednisolone lowered the rates of bone resorption and formation in OVX rats. The osteopenic effects of prednisolone and ovariectomy were additive and independent. E2 protected bone from the osteopenic effects of ovariectomy but did not affect bone loss induced by prednisolone. These results suggest prophylactic estrogen should help to avoid bone loss from estrogen deficiency in patients requiring chronic high dose glucocorticoid treatment.     

Estrogen modulates the recruitment of myelopoietic cell progenitors in rat through a stromal cell-independent mechanism involving apoptosis

Loss of ovarian function leads to a significant increase in the number of bone-resorbing osteoclasts. Estrogen replacement is known to manifest bone protective effects in the treatment of postmenopausal osteoporosis. In the present study, we used ovariectomized rats to examine the effects of estrogen loss at the osteoclast progenitor colony forming unit-granulocyte macrophage (CFU-GM) level. A significant increase in CFU-GM number was observed as early as 7 days following ovariectomy, and correlated directly with an increase in the number of osteoclast-like cells generated in marrow cultures. The increase in CFU-GM following ovariectomy was abrogated in animals that received estrogen treatment in vivo. A similar suppressive effect was observed on CFU-GM number when ovariectomized rat marrow was treated with estrogen in vitro. This effect was blocked in the presence of the estrogen antihormone ICI 164,384. Thus, the data suggest the possibility that estrogen exerts a direct effect on osteoclast progenitors, and does so through the estrogen receptor-mediated mechanism. Ovariectomy also led to an increase in the early hematopoietic stem/progenitor cell population (Thy 1.1+ cells) as determined by FLOW cytometry methods. Morphological changes as well as terminal deoxynucleotidyl transferase assays revealed that estrogen treatment negated growth factor-induced proliferation of these early progenitors by promoting apoptosis. The cellular effects of estrogen in vitro together with the immunocytochemical detection of the estrogen receptor in these cells, strongly support the contention that in addition to osteoclast progenitors such as CFU-GM, earlier hematopoietic progenitors are also unique cellular targets for estrogen action.     

Ovariectomy augments hypertension in aging female Dahl salt-sensitive rats

The ovariectomized (OVX) Dahl salt-sensitive (DS) rat fed a low-salt diet is a model of postmenopausal hypertension. In addition to estrogen loss, aging can also contribute to postmenopausal hypertension. We hypothesized that: (1) female DS rats on a low-salt diet become hypertensive with age; (2) ovariectomy accelerates age-dependent hypertension in the DS rat caused by estrogen depletion; and (3) this hypertension correlates with increased type 1 angiotensin receptor (AT1R) number (Bmax). Blood pressure was monitored by telemetry from 3 to 12 months and AT1R Bmax was determined by Scatchard analysis in glomeruli and adrenal cortex. Three groups of DS rats were studied: intact, OVX, and 17beta-estradiol-replaced OVX (OVX+E). In intact rats, aging to 12 months resulted in hypertension (159+/-6 mm Hg) and an 82% decrease in estrogen. Blood pressure in OVX was significantly higher than OVX+E through 12 months of age (173+/-4 versus 150+/-8 mm Hg). At 4 months, OVX increased AT1R Bmax compared with intact and OVX+E in both glomeruli and adrenal cortex. Aging also increased AT1R Bmax in these tissues in intact rats. In summary, female DS rats fed a low-salt diet have hypertension develop with age, that is accelerated by OVX and attenuated by estrogen replacement. Concurrently, AT1Rs are upregulated by age and OVX, which is prevented by estrogen replacement. This study suggests that an increased activity of the renin angiotensin system contributes to the development of hypertension, and estrogen protects against this process.     

Ovarian hormone modulates 5-hydroxytryptamine 3 receptors mRNA expression in rat colon with restraint stress-induced bowel dysfunction

AIM: To examine the effects of ovarian hormone on the expression of 5-hydroxytryptamine 3 receptors (5-HT3R) in rat colon of restraint stress-induced bowel dysfunction. METHODS: Twenty-four female Sprague-Dawley rats were randomly divided into three groups of 8 each: sham operation, ovariectomy (OVX) and ovariectomy with estrogen (E2) and progesterone (P) replacement therapy (OVX+E2+P). The rats were subjected to 1-h restraint stress 4 wk after operation. The changes of defecation were monitored by collection of fecal pellets. The gonadal steroids were measured in duplicate by radioimmunoassay (RIA). The expression of 5-HT3R mRNA in the colon was studied by RT-PCR. RESULTS: Compared with sham group and OVX+E2+P group, OVX group showed increase in fecal pellets and decrease in the time of vitreous pellets excretion (P<0.01). Serum levels of E2 and P were suppressed in OVX group and restored following treatment with ovarian steroids (P<0.01), and the levels of 5-HT3R mRNA in the colon of ovariectomized rats were significantly increased, the expression of 5-HT3R mRNA was significantly decreased in hormone replacement therapy group (P<0.01). CONCLUSION: Ovarian hormone plays a role in the regulation of 5-HT3R expressions in restraint stress-induced bowel dysfunction of rats. The interactions between ovarian steroids and gastrointestinal tract may have major pathophysiological implications in 5-HT-related disorders, such as irritable bowel syndrome (IBS).     

Increased B-lymphopoiesis by interleukin 7 induces bone loss in mice with intact ovarian function: Similarity to estrogen deficiency

Estrogen deficiency caused by ovariectomy (OVX) results in a marked bone loss due to stimulated bone resorption by osteoclasts. During our investigations of the pathogenesis of bone loss in estrogen deficiency, we found that OVX selectively stimulates B-lymphopoiesis which results in marked accumulation of B220-positive pre-B cells in mouse bone marrow. To examine the possible correlation between stimulated B-lymphopoiesis and bone loss, 8-week-old female mice were treated with interleukin (IL) 7, which stimulates B-lymphopoiesis in bone marrow. We also examined bone mass in IL-7 receptor-knockout mice that exhibit marked suppression of B-lymphopoiesis in the bone marrow. The increased B-lymphopoiesis induced by IL-7 administration resulted in marked bone loss by stimulation of osteoclastic bone resorption in mice with intact ovarian function. The changes in both B-lymphopoiesis and bone mass in IL-7-treated female mice were similar to those in age-matched OVX mice. In contrast, the trabecular bone volume of the femur was greatly increased in both female and male IL-7 receptor-knockout mice when compared with the respective wild-type and heterozygous littermates. These results show that the perturbation of B-lymphopoiesis in the bone marrow is closely linked to the change in bone mass. We propose here that the increased B-lymphopoiesis due to estrogen deficiency is involved in the mechanism of stimulated bone resorption.     

雌激素对去卵巢大鼠内脏脂肪细胞脂肪因子表达水平的影响

目的 观察雌激素对去卵巢大鼠内脏脂肪细胞瘦素、脂联素、抵抗素和肿瘤坏死因子-α(TNF-α)表达水平的影响,探讨雌激素对体脂分布的影响机制.方法 6周龄Sprauge-Dawley雌性大鼠30只,采用随机数字表法分成3组:假手术组、去卵巢组和去卵巢+戊酸雌二醇组(OVX+E2组),每组10只.术后1周,OVX+E2组大鼠每天按1 mg/kg体重灌胃戊酸雌二醇水溶液,其他组大鼠灌胃等体积蒸馏水.连续给药12周后,腹主动脉取血,迅速剥离内脏脂肪组织.采用全自动生化分析仪检测血脂、血糖.采用免疫组化染色、实时荧光定量RT-PCR和Western印迹检测脂肪细胞瘦素、脂联素、抵抗素和TNF-α的表达.结果 3组血清瘦素、脂联素和抵抗素水平差异无统计学意义(P均＞0.05),但去卵巢组TNF-α水平显著高于假手术组(F=4.785,P＜0.05).免疫组化显示,与假手术组相比,去卵巢组内脏脂肪组织中瘦素表达明显减弱,而脂联素、抵抗素和TNF-α表达明显增强;与去卵巢组相比,OVX+E2组内脏脂肪组织中瘦素表达明显增强,脂联素、抵抗素和TNF-α表达明显减弱(F =3.712 ～5.198,P均＜0.05).3组内脏脂肪细胞瘦素mRNA和蛋白表达水平差异无统计学意义(P均＞0.05);去卵巢组内脏脂肪细胞脂联素、抵抗素和TNF-α的mRNA和蛋白表达水平显著高于假手术组,而OVX+E2组内脏脂肪细胞脂联素、抵抗素和TNF-α的mRNA和蛋白表达水平显著低于去卵巢组(F=3.175～5.342,P均＜0.05).结论 雌激素可通过下调去卵巢大鼠内脏脂肪细胞脂联素、抵抗素和TNF-α的表达,进而影响去卵巢大鼠体脂再分布.     

Estrogen treatment prevents osteopenia and depresses bone turnover in ovariectomized rats

Female Sprague-Dawley rats were subjected to bilateral ovariectomy (OVX) or sham surgery (control). Groups of ovariectomized (OVX) and control rats were injected daily with low, medium, or high doses of 17 beta-estradiol (10, 25, or 50 micrograms/kg BW, respectively). An additional group of OVX and control rats was injected daily with vehicle alone. All rats were killed 35 days after OVX, and their proximal tibiae were processed undecalcified for quantitative bone histomorphometry. Trabecular bone volume was markedly reduced in vehicle-treated OVX rats relative to that in control rats (12.1% vs. 26.7%). This bone loss was associated with a 2-fold increase in osteoclast surface and a 4-fold increase in osteoblast surface. The bone formation rate, studied with fluorochrome labeling, was also significantly elevated in vehicle-treated OVX rats (0.111 vs. 0.026 micron3/micron2.day). In contrast, treatment of OVX rats with the three doses of estradiol resulted in normalization of tibial trabecular bone volume and a decline in histomorphometric indices of bone resorption and formation. Our results indicate that estrogen treatment provides complete protection against osteopenia in OVX rats. The protective mechanism involves estrogenic suppression of bone turnover. These findings are consistent with the skeletal effects of estrogen therapy in postmenopausal women.     

仙灵骨葆对去势大鼠骨量和生物力学性能的影响

目的探讨仙灵骨葆对骨质疏松(OP)大鼠骨量、骨代谢和生物力学性能的影响。方法 3月龄雌性SD大鼠24只分为3组,每组8只:正常对照组(N)、卵巢切除组(OVX)、卵巢切除+仙灵骨葆治疗组(XLGB)。除N组外,其余两组行卵巢切除术,6 w后XLGB组给予药物干预:250 mg.kg-1.d-1,OVX组给予等量生理盐水,8 w后处死所有大鼠。留取尿液、血清检测血PINP值、尿DPYD/Cr、NTX/Cr值。取左侧股骨行骨密度测定,取左侧胫骨制备硬组织不脱钙切片,备行骨组织形态计量学检测,取右侧股骨行三点弯曲试验,检测其最大载荷。结果 OVX组血PINP、尿DPYD/Cr、尿NTX/Cr值显著高于N组,XLGB能显著降低血PINP、尿DPYD/Cr、尿NTX/Cr值,但仍显著高于N组。OVX组股骨全长及近、中、远三段骨密度均显著低于N组,XLGB组近、远端骨密度显著高于OVX组。BV/TV在OVX组显著低于N组,XLGB组显著高于OVX组;OVX、XLGB组骨吸收指标Oc.N、Er.Pm均显著高于N组,XLGB组Oc.N、Er.Pm显著低于OVX组,BFR/BV显著高于OVX组。最大载荷三组之间无显著差别。结论仙灵骨葆灌胃可抑制卵巢切除大鼠骨量丢失,其机制与促进骨形成、抑制骨吸收,降低骨转换水平,进而维持骨量及微观结构有关。