Ovariectomy in the rat induces a rapid increase in the urinary excretion of hydroxylysine glycosides and non-reducible crosslink residues

The ovariectomized rat is the most commonly used animal model of human postmenopausal osteoporosis, exhibiting a high rate of bone turnover with resorption exceeding formation. At present, bone turnover is quantified directly by dynamic histomorphometry. The aim of the present study was to determine whether the measurement of the urinary output of some specific bone collagen catabolites — pyridinolines and hydroxylysine glycosides — could be used to indirectly monitor the initial phase of bone turnover increase in ovariectomized 90-day-old rats. Ninety-day-old female rats were randomly divided into three groups (n=6): ovariectomized, sham-operated and non-treated controls. Urine samples (24 h) were collected 6 days before surgery and twice weekly for the 4 weeks following ovariectomy. Urinary excretion of pyridinoline (PYD), deoxypyridinoline (DPD), glucosyl-galactosyl-hydroxylysine (GGHYL) and galactosyl-hydroxylysine (GHYL) were measured. As expected, ovariectomy was associated with a significant decrease in bone mineral density in both the proximal tibial and distal femoral metaphysis. Compared with both sham-operated and control animals, ovariectomized rats showed significant increases in PYD, GGHYL and GHYL urinary output 8 days after surgery and in DPD output after 15 days. These changes were maintained throughout the study. The results confirm that measurement of the urinary excretion of pyridinolines and hydroxylysine glycosides represents a powerful tool for detecting the onset of bone turnover in ovariectomized 90-day-old rats.     

1α-Hydroxyvitamin D2 Partially Dissociates Between Preservation of Cancellous Bone Mass and Effects on Calcium Homeostasis in Ovariectomized Rats

Vitamin D metabolites can prevent estrogen depletion-induced bone loss in ovariectomized (OVX) rats. Our aim was to compare the bone-protective effects of 1α,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃), 1α,25-dihydroxyvitamin D₂ (1,25(OH)₂D₂), 1α-hydroxyvitamin D₃ (1α(OH)D₃), and 1α-hydroxyvitamin D₂ (1α(OH)D₂) in OVX rats. 1α(OH)D₃ and 1α(OH)D₂ are thought to be activated in the liver to form 1,25(OH)₂D₃ and 1,25(OH)₂D₂, respectively. Forty-four 12-week-old female Fischer-344 rats were either OVX or sham-operated (SHAM). Groups of OVX rats (n = 7 each) received vehicle alone, 1,25(OH)₂D₃, 1,25(OH)₂D₂, 1α(OH)D₃, or 1α(OH)D₂, starting 2 weeks after surgery. All vitamin D metabolites were administered orally at a dose of 15 ng/day/rat. Urine and blood samples were collected 6, 9, 12, and 16 weeks after surgery. Serum samples were analyzed for total calcium and phosphate. Calcium, phosphate, creatinine, and free collagen cross-links (ELISA) were determined in urine. After tetracycline double labeling, the rats were sacrificed 16 weeks postsurgery, and the proximal tibiae and the first lumbar vertebrae were processed undecalcified for static and dynamic bone histomorphometry. 1,25(OH)₂D₃ and, to a slightly lesser extent, 1,25(OH)₂D₂ elevated vertebral cancellous bone mass in OVX rats to a level beyond that observed in SHAM animals, and both compounds increased serum calcium and urinary calcium excretion to similar extents. 1α(OH)D₃ and 1α(OH)D₂ resulted in a 64% and 84%, respectively, inhibition of ovariectomy-induced vertebral cancellous bone loss. In the proximal tibial metaphysis, all vitamin D metabolites tested could only partially prevent post-OVX trabecular bone loss, with a tendency for 1α(OH)D₃ to be the least active compound. The effects of 1α(OH)D₃ and 1α(OH)D₂ on calcium homeostasis differed markedly, however. The mean increase in urinary calcium excretion over the whole experiment was fivefold for 1α(OH)D₃, whereas the corresponding increase for 1α(OH)D₂ was only twofold. We conclude that, compared with 1α(OH)D₃, 1α(OH)D₂ combined at least equal or higher bone-protective activity in OVX rats with distinctly less pronounced effects on calcium homeostasis. This effect was not due to a differential action of the corresponding main activation products, 1,25(OH)₂D₃ and 1,25(OH)₂D₂. Received: 2 May 1996 / Accepted: 18 October 1996     

Effects of Clodronate on Cortical and Trabecular Bone in Ovariectomized Rats on a Low Calcium Diet

The aim of this study was to evaluate the contribution of a low calcium diet to the cortical and trabecular osteoporosis seen in ovariectomized rats after 7 weeks on a low calcium diet and to investigate the effects of the bisphosphonate clodronate on this development of osteoporosis. Thirty-six mature, female Wistar rats were randomized into four groups: Ovx−B (bisphosphonate) and Ovx−C (control) were ovariectomized, and Sham−Ca (low calcium) and Sham+Ca (normal calcium) were sham operated. The first three groups were fed a low calcium diet (0.01%) and Sham+Ca normal rat chow (Ca 1.1%). The Ovx−B received 10 mg/kg s.c. clodronate daily for nine weeks, and Ovx−C, Sham−Ca, and Sham+Ca received the same volumes of saline. Bone mineral turnover measured as ⁸⁵Sr-uptake was increased in all low calcium groups compared to Sham+Ca. The Sham+Ca femora had higher dry weight and ash weight than the other groups, and Ovx−C had higher dry weight compared with Ovx−B and Sham−Ca. Calcium content was lower in both Ovx groups compared to both Sham groups. Magnesium was lower in all groups compared to Sham+Ca and higher in Ovx−B compared with Ovx−C. In the femoral shaft, Sham+Ca had significantly higher ultimate bending moment, energy absorption, and deflection compared to the other three groups. Ultimate bending moment was higher in Sham−Ca than in Ovx−C. Stiffness was increased in both Sham+Ca and Ovx−B compared to Ovx−C. The maximum stress in the femoral midshaft was higher in Sham+Ca than in the other groups, and higher in Ovx−B than in Ovx−C. Histomorphometry showed increased medullary area in all low calcium groups compared to Sham+Ca and larger cortical area in Sham+Ca and Ovx−B compared to Ovx−C. Compared to Sham+Ca the trabecular bone volume was decreased to 30% in Sham−Ca and to 9% in Ovx−C, but was unchanged in Ovx−B. The low calcium diet generally increased bone mineral turnover and reduced the tibial bone volume. Femoral changes led to a reduction of cortical fracture strength and maximal stress. Ovariectomy in addition to a low calcium diet reduced femoral strength even more. Daily injections of clodronate to ovariectomized rats on a low calcium diet increased femoral shaft stiffness and maximum stress, and clodronate preserved both trabecular and cortical tibial bone volume completely. Received: 11 June 1996 / Accepted: 5 March 1997     

中药骨康对去卵巢大鼠腰椎骨形态计量学的影响

目的 探讨中药骨康对卵巢切除大鼠骨质疏松症的治疗作用。方法 选择6m龄SD大鼠48只。随机分为假手术模型组（Sham）、手术模型组（OVX）、尼尔雌醇组（E2）、中药骨康组。切除大鼠卵巢3m后。大鼠骨质疏松症模型制备成功，分别给予尼尔雌醇、中药骨康灌胃治疗，并与Sham组和OVX组对照。治疗3m后，体内双荧光标记，取第2腰椎包埋切片。全自动图像分析及松质骨骨形态计量学软件处理。观察中药对骨形态计量学参数的影响。结果 卵巢切除后大鼠骨小梁面积百分数（％Tb．Ar）下降35．84％，骨小梁数量（Tb．N）下降16．60％，骨小梁宽度（Tb．Th）下降25．79％，表明绝经后骨质疏松症动物模型成立。骨康治疗3个月后，与OVX组相比，Oc．N／mm^2下降42．80％，有显著性差异（P〈0．01）；％Tb．Ar、Tb．Th、Tb．N和MAR有上升趋势（P〉0．05）；15．Sp，％L．Pm、BFR／BS、BFR／TV和Oc．N／mm，BFR／BV有下降趋势（P〉0．05）。结论 中药骨康具有降低骨转换以及促进骨形成和抑制骨吸收的双重作用，说明中药骨康对骨质疏松有明显的治疗作用。     

App17肽防治去卵巢大鼠海马神经细胞的凋亡

目的:观察去卵巢大鼠几种神经元凋亡相关蛋白的表达,并用TUNEL试剂盒检测,研究缺乏雌激素是否引起神经元凋亡;评估App17肽是否能改善去卵巢大鼠的神经细胞凋亡,初步探讨App17肽的神经保护作用机制。方法:雌性Wistar大鼠随机分3组:假手术组(shamcontrol组),卵巢去势对照组(OVX组),App17肽实验组(17P+OVX组)。Sham组做假手术,OVX组和17P+OVX组做双侧卵巢切除术。17P+OVX组从卵巢去势第7周开始用App17肽治疗6周,用免疫组化和Westernblot方法观察AIF、Bax、Bcl-2的表达,用TUNEL法检测凋亡情况。结果:免疫组织化学和Westernblot结果表明App17肽实验组AIF、Bax表达明显少于去卵巢组;Bcl-2在App17肽实验组的表达明显高于去卵巢组。TUNEL结果表明App17肽实验组凋亡细胞明显少于去卵巢组。结论:雌激素缺乏引起去卵巢大鼠海马和皮层神经细胞凋亡相关蛋白改变和出现凋亡细胞,App17肽具有明显的防治作用。     

卵巢切除对大鼠子宫雌激素受体亚型表达的影响

目的 通过对去卵巢大鼠雌激素受体亚型在子宫表达的观察，研究卵巢切除对大鼠子宫影响的机制。方法 选择成年Wistar大鼠30只，10只为正常组，10只为假手术组，10只为模型组(去卵巢组)。6周后处死大鼠，分离摘取子宫，常规石蜡包埋、切片，分别进行ERα、ERβ抗体的免疫组织化学染色。结果 1．ERα在正常组、假手术组及模型组子宫的上皮细胞、间质细胞及肌细胞核均有表达，其中以腺上皮细胞表达最强。图像分析表明，模型组腺上皮ERα表达较其他2组弱。2．ERα在去卵巢子宫中未见明显表达，只在正常组和假手术组子宫腺上皮细胞核中表达，且较ERα表达弱。结论卵巢切除后明显影响大鼠子宫ERα、ERα的表达，尤其对ERα的影响较为显著。     

17βE2对切除卵巢兔动脉粥样硬化与血液流变学的作用

目的: 探讨17βE₂对卵巢切除(OVX)兔的AS斑块、血脂代谢及血液流变学的影响。方法: 34只成熟未孕雌兔分为4组:A为NC;B为Sham+CHO;C为OVX+CHO;D为OVX+CHO+17βE₂。喂养12周,喂养前与12周后测定血脂TC、TG、HDL-C、LDL-C、ApoA1、ApoB;测定全血粘度、血浆粘度、AIRC及纤维蛋白原。喂养12周后处死,测定AS斑块面积与主动脉总面积百分比。结果: ①AS斑块面积与主动脉总面积之比,A、B、C、D组分别为0.02±0.00、0.42±0.15、0.67±0.23、0.12±0.11,B组小于C组(P＜0.05)、D组明显小于C组(P＜0.01)。②血脂TC、TG、LDL-C、ApoB,B、D组明显小于C组(P＜0.01)。③血液流变学:全血粘度、血浆粘度AIRC与纤维蛋白原D组明显低于C组(P＜0.01)。结论: 17βE₂能抑制脂质斑块沉积,改善血脂代谢,减轻高脂血症,改善血液流变学。这可能是雌激素抑制AS形成的部份机理。     

Genetic obestiy: estrogenic influences on the body weight and food intake of lean and obese adult Zucker (fa/fa) rats

The effects of chronic estrogen withdrawal and subsequent hormone replacement on the feeding and body weight of adult lean and genetically obese Zucker rats were investigated. Following confirmation of a delay in the vaginal canalization of the fatty rat, subgroups of each genotype received either ovariectomy or sham surgery (Experiment 1). One hundred days later all subjects were injected subcutaneously (SC) with 1.0 microgram of estradiol benzoate (EB) daily for 16 treatment days (Experiment 2A). A second series of daily 2.0 microgram EB injections was administered intraperitoneally (IP) for 1 week (Experiment 2B). The first experiment revealed that ovariectomy produced overeating and similar weight gains in both genotypes. In the second experiment, SC hormone treatment completely reversed ovarian obesity in lean animals but failed to alter the food intake or weight gain of fatty rats. IP administration of EB depressed the feeding of fatty and lean animals to a comparable degree but a reduction in weight gain was observed only in the lean rats. These findings are discussed in light of current theories of estrogenic modulation of energy balance.     

Organization and expression of agonistic and socio-sexual behavior in golden hamsters over the estrous cycle and after ovariectomy

Paris of hamsters were housed in large enclosures that contained separate male and female living areas and observed over the 4-day estrous cycle and after ovariectomy. Agonistic elements exhibited frequently by females included on-back, boxing, lateral posturing, and biting, whereas males engaged frequently in boxing and on-back patterns of behavior. Furthermore, on-back and boxing by females were significantly higher on estrus than on any other day of the estrous cycle. Agonistic acts performed after ovariectomy did not differ in occurrence from those shown by animals on diestrus and proestrus. Vaginal marking increased during diestrus and attained a peak 24 hr prior to sexual receptivity. Both vaginal marking and mating occurred more frequently in the female's than male's home area suggesting that vaginal marks on days preceding behavioral estrus serve to attract males to the nest of females. Males also organized their marking patterns by location as shown by more flank marking in their own than their partner's area, albeit the significance for this difference in location is not known. The results demonstrate that when heterosexual paris of hamsters are tested in large and partially familiar habitats, a wide range of behavior is exhibited and organized in a manner that is not observed in small and unfamiliar cages.