己烯雌酚对去卵巢大鼠密质骨影响的定量研究

将２７只３月龄ＳＤ雌大白鼠随机分为基础对照组（Ａ）、年龄对照组（Ｂ）、去卵巢组（Ｃ）和去卵巢加己烯雌酚（ＤＥＳ）治疗组（Ｄ）。Ｂ组和Ｃ组用生理盐水５ｍｌ·ｋｇ－１／ｄ．ｉｇ，Ｄ组用浓度为４．５ｍｇ／Ｌ的己烯雌酚按５ｍｌ·ｋｇ－１／ｄ．ｉｇ，每周６次。１２周后，对各组大鼠胫骨中段不脱钙骨片进行骨计量学分析。去卵巢大鼠由于骨吸收大于骨形成，密质骨变薄，骨髓腔扩大，出现高转换型骨质疏松，己烯雌酚能明显抑制去卵巢后的骨高转换，保持密质骨厚度，维持骨量的正常。     

氟化钠对去卵巢大鼠胫骨作用的骨形态计量学研究

目的 了解卵巢切作大鼠胫骨对氟化钠的治疗反应。方法 将２４只雌性大鼠随机分成３组,单纯切卵巢组、对照组和氟化钠治疗组。对单纯切卵巢组和氟化钠治疗组大鼠行卵巢切除手术,建立骨质疏松动物模型,氟化钠治疗组于卵巢切除术后１个月予氟化钠,给药３个月后处死。用骨形态计量学方法检测卵巢切除大鼠无后肢胫骨对氟化钠的治疗反应。结果 单纯卵巢切除组大鼠骨小梁体积,平均骨小梁密度较对照组显著较减少（Ｐ〈０．０５）,四     

何首乌煎剂对去卵巢大鼠骨质丢失的防治作用

目的探讨何首乌煎剂对去卵巢大鼠骨丢失的骨组织形态计量学改变及预防作用。方法大鼠双侧卵巢去除术(OVX)后预防用药90d。骨标本行不脱钙骨制片,骨组织形态计量学测量胫骨近心端松质骨静态和动态参数。结果去卵巢大鼠骨小梁面积百分率(%Tb.Ar)减少(P<0.05),出现高转换型骨质疏松:己烯雌酚可完全对抗去卵巢大鼠的体重增加、骨转换率增高和骨量丢失;何首乌煎剂使去卵巢大鼠的%Tb.Ar增加(P<0.05)、骨吸收减少、骨转换率呈下降趋势,不抑制藕联的骨形成。结论何首乌煎剂对去卵巢大鼠骨丢失有一定的预防作用,且对子宫无明显刺激。     

尼尔雌醇对去卵巢大鼠骨代谢影响的定量研究

实验用３月龄雌ＳＤ大鼠，分基础对照组、年龄对照组、去卵巢组及去卵巢加尼尔雌醇治疗组（ＣＥＥ１ｍｇ·ｋｇ－１，每周ｉｇ１次），１２ｗ后，对各组大鼠胫骨近端不脱钙骨进行骨组织形态计量学分析。结果表明：去卵巢大鼠骨形成和骨吸收均明显增加，但骨吸收大于骨形成，出现高转换型骨质疏松。尼尔雌醇通过明显抑制去卵巢大鼠的骨吸收和骨形成，阻止骨高转换，保持骨量的正常。     

骨灵片对去卵巢骨质疏松大鼠腰椎骨形态计量学及生物力学的影响

目的观察补肾中药——骨灵片对去卵巢（OVX）骨质疏松（OP）大鼠腰椎的骨组织形态计量学以及生物力学影响,进一步探讨补肾中药治疗OP的机制。方法选择3月龄雌性SD大鼠36只,随机分为假手术组、OVX组、骨灵片低、中、高剂量组和雌激素组,每组各6只。试验4个月后采用骨形态计量学检测腰椎骨的动静态参数,生物力学凹入法检测腰椎的最大载荷和刚度。结果①与OVX组比较,骨灵片各剂量组和雌激素组的骨小梁分离度、骨形成率（BFR/BS、BFR/BV）、每mm破骨细胞数均有显著下降（P〈0.01）;骨灵片中剂量组BFR/TV和中高剂量组的骨矿化沉积率（MAR）减少有显著差异（P〈0.01）;骨灵片中高剂量组和雌激素组骨小梁面积百分数、厚度和数量显著升高且中剂量组的骨小梁厚度比雌激素组有显著增加（P〈0.05）;②生物力学上,与去卵巢组比较,雌激素组和骨灵片各剂量组的刚度以及骨灵片中高剂量组的最大载荷均有显著升高（P〈0.01）。骨灵片高剂量组在最大载荷上的提高较雌激素组有显著差异（P〈0.05）。结论补肾中药骨灵片具有促进骨形成和抑制骨吸收的双重作用,降低骨转换,使骨结构得到改善,并明显增加骨生物力学强度。     

强骨宝对去卵巢大鼠骨生物力学及骨量指标的影响

目的观察强骨宝复方制剂干预去卵巢骨质疏松模型大鼠股骨骨生物力学性能参数和骨矿元素钙、镁、磷及羟脯氨酸含量的变化,并探讨两者变化的相互关系。方法取32只Wistar雌性大鼠随机分为4组,按要求行去势手术后分别灌胃给药,8w后处死,取大鼠左侧股骨进行三点弯曲实验,测定股骨生物力学性能参数;取右侧股骨烘干测定钙、镁、磷及羟脯氨酸含量。结果强骨宝可使去卵巢大鼠股骨生物力学性能参数结构和材料力学指标均显著增加（P〈0.01,P〈0.05）;骨干重、羟脯氨酸、钙、镁等骨量指标亦明显提高（P〈0.01,P〈0.05）。结论强骨宝能促进骨羟脯氨酸的合成,钙、镁、磷的吸收和沉积,导致骨量增加;同时能促进骨的结构和材料力学特性的改善,增加骨对外界应力的对抗作用,而且骨生物力学参数与骨量指标两者的变化统一。     

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。     

氟与羟乙膦酸钠联合用药对去卵巢大鼠骨计量学和生物力学性质的影响

目的评价抗骨吸收药物与促进骨形成药物联合应用对去卵巢大鼠骨质疏松的潜在治疗作用。方法观察单独用羟乙膦酸钠（１ｍｇ·ｋｇ－１·ｄ－１）和氟钙剂（０４５ｎｇＦ－＋１３５６ｍｇＣａ２＋）·ｋｇ－１·ｄ－１与两者联合用药对去卵巢大鼠胫骨近端次级松质骨骨计量学指标和股骨中段骨生物力学性质的影响。结果与去卵巢不处理组比较，去卵巢氟钙剂处理组、去卵巢羟乙膦酸钠处理组及两者联合用药组小梁骨面积百分比分别高出４２％、１０９％、１１０％；小梁骨的数量分别多２４％、９４％、９０％；小梁骨分离度分别低３５％、１４８％、１３８％；吸收侵蚀表面分别少４５％、５０％、６３％；刺激频率在后两组分别低６１％、５７％。与假手术组比较，上述三种处理使矿化延迟时间分别延长１６％、４２％、２９％；联合用药组股骨干力学强度明显增加。上述诸参数的改变差异均有显著性（Ｐ＜００５）。结论羟乙膦酸钠能显著抑制骨吸收，降低骨转换，其保护小梁骨显微结构作用优于单用氟钙剂处理，与氟钙剂合用明显增加股骨的力学强度。但三种处理均有潜在的抑制矿化作用     

去卵巢大鼠股骨和腰椎骨生物力学特性及骨矿物含量的变化

目的比较去卵巢大鼠股骨和腰椎的骨生物力学特性及骨矿物含量的不同。方法10.5月龄未交配的雌性SD大鼠40只,随机分为10.5月龄基础对照组;13月龄假手术组;13月龄去卵巢10 w组;16月龄假手术组;16月龄去卵巢22 w组。大鼠行双侧卵巢去除术后,然后灌喂生理盐水5 m l.kg-1.d-1。大鼠处死后采用三点弯曲试验测定股骨和压缩试验测定腰椎的骨生物力学指标,然后分别测量股骨和腰椎的骨矿物质含量。结果去卵巢大鼠腰椎的骨破坏荷载、破坏应力、弹性模量均明显降低及骨钙、骨磷、骨镁含量均明显降低,但股骨生物力学参数和矿物质含量均无明显变化。结论去卵巢大鼠腰椎的骨生物力学和骨矿物含量均明显降低,而股骨变化不明显。     

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.     

红花水溶性成分抗氧化作用的研究

本文观察了红花水提液及其醇沉所得组分的抗氧化作用，光谱扫描及TLC分析结果表明，红花黄色素主要集中于醇沉所得沉淀中，上清液仅存少量极性较强的红花黄色素。邻二氮菲-Fe2+氧化法及硫代巴比妥酸比色法检测结果表明，红花水提液可清除羟自由基，抑制小鼠肝匀浆脂质过氧化。以上述两指标比较研究的结果表明，水提液醇沉后抗氧化活性主要集中于醇沉所得红花黄色素中，提示红花水溶性抗氧化有效组分可能为红花黄色素。     

Separate and combined effects of growth hormone and parathyroid hormone on cortical bone osteopenia in ovariectomized aged rats.

The focus of this study is on whether cortical osteopenia occurs in ovariectomized aged female rats, and if so, whether growth hormone (GH) and parathyroid hormone (PTH) independently or together (GH+PTH) can rebuild the lost cortical bone. Tibio-fibula junction was analyzed by histomorphometry and peripheral quantitative computerized tomography (pQCT) densitometry. Significant loss of cortical bone area (Ct. BAr), cortical bone mineral content (Ct. BMC), cortical thickness (Ct. Th) and increase of endocortical perimeter occurred 4 months after ovariectomy. The rats were given GH, PTH, GH+PTH or vehicle for 2 months and sacrificed. GH, PTH and GH+PTH increased Ct. BAr, Ct. BMC, Ct. Th, periosteal perimeter, periosteal double-labeled perimeter, mineral apposition rate, and bone formation rate, but decreased marrow area. PTH and GH+PTH decreased endocortical perimeter, and increased endocortical double labeled perimeter and bone formation rate. In conclusion, ovariectomy induced cortical bone loss in aged rats by increasing endocortical bone resorption. Growth hormone increased periosteal bone formation, while PTH stimulated endocortical bone formation and in combination GH+PTH produced complementary effects thereby reversing osteopenia.     

Endocrine and pharmacological suppressors of bone turnover protect against osteopenia in ovariectomized rats

This study was designed to test the hypothesis that endocrine and pharmacological suppressors of bone turnover prevent the development of osteopenia during estrogen deficiency. Sham-operated control and ovariectomized (OVX) rats were treated intermittently with vehicle alone, estrogen, or the diphosphonate compounds etidronate disodium (EHDP) and NE-58095 [2-(3-pyridinyl)2-hydroxyethylidene-1,1-bisphosphonate disodium] for 35 or 70 days after surgery. Their proximal tibiae were processed undecalcified for quantitative bone histomorphometry. Vehicle-treated OVX rats were characterized by decreased cancellous bone volume and 3- to 4-fold increases in osteoblast surface, osteoclast surface, bone formation rate, and bone resorption rate. Treatment of OVX rats with estrogen and NE-58095 provided complete protection against bone loss and significantly depressed all of the above indices of bone turnover. OVX rats treated with EHDP exhibited at least partial protection against bone loss and decreased bone turnover. EHDP induced a mild mineralization defect, as indicated by a prolonged mineralization lag time at the tibial endocortical surface. The new diphosphonate compound NE-58095 did not impair bone mineralization. Our results indicate that endocrine and pharmacological suppressors of bone turnover prevent the development of osteopenia during the early stages of estrogen deficiency. If confirmed by clinical trials in humans, diphosphonate compounds may prove to be an alternative to estrogen for the prevention of postmenopausal bone loss.     

Sevelamer restores bone volume and improves bone microarchitecture and strength in aged ovariectomized rats

Sevelamer hydrochloride, a noncalcium phosphate binder, has been shown to reduce coronary artery and aortic calcification, and to improve trabecular bone mineral density in hemodialysis patients with chronic kidney disease. Here, we examined whether sevelamer given orally for 12 wk with normal food could restore bone volume (BV) and strength in aged ovariectomized (OVX) rats starting at 4 wk after OVX. Dual-energy x-ray absorptiometry, microcomputerized tomography, and bone histomorphometry analyses showed that OVX animals receiving sevelamer had increased trabecular BV (51%), trabecular number (43%), trabecular thickness (9%), cortical thickness (16%), mineral apposition rate (103%), bone formation rate (25%), and enhanced cortical and trabecular bone mechanical strength as compared with OVX rats. Sevelamer decreased collagen C telopeptide, increased osteocalcin levels, and decreased phosphate and magnesium levels without affecting calcium levels in the blood. Although sevelamer was not absorbed systemically, it stimulated osteoblast differentiation in BM-derived mesenchymal stem cell cultures, as evaluated by alkaline phosphatase positive colony-forming units, and inhibited recombinant human soluble receptor activator of nuclear factor-kappaB ligand-induced osteoclast differentiation, as evaluated by tartrate-resistant acid phosphatase positive cells in bone mineral-hematopoietic stem cell cultures. Surface enhanced laser desorption/ionization time-of-flight mass spectrometry analysis revealed that 69 proteins were differently expressed after OVX, of which 30% (20 of 69) were reversed to sham activity after sevelamer intake. PTH, fibroblast growth factor-23, and cytokine profile in serum were not significantly changed. Together, these results suggest that sevelamer in food increases the BV and improves biomechanical properties of bone in OVX rats.     

抗骨松颗粒对去卵巢大鼠骨质含量和生物力学的影响

目的 探讨抗骨松颗粒对去卵巢大鼠骨质疏松模型骨质含量和骨生物力学性能的影响.方法 40只10月龄Wistar雌性大鼠随机分为对照组、模型组、抗骨松组和倍美力组,每组10只.正常对照组做假手术,其余3组做卵巢切除术.术后3个月开始给药,连续用药满90 d,处死动物,取出第2腰椎和股骨,测定骨密度、钙、磷、锰、镁、羟脯氨酸、碱性磷酸酶和有机质含量;取出第3腰椎,测骨质含量和生物力学性能.结果 抗骨松颗粒能明显提高骨质疏松大鼠的骨密度、钙、锰、镁、羟脯氨酸和有机质含量,增强腰椎骨的抗压生物力学性能.结论 应用抗骨松颗粒可以明显改善去卵巢大鼠骨质疏松模型骨质含量和骨的生物力学性能.     

Total administered dose of ibandronate determines its effects on bone mass and architecture in ovariectomized aged rats

OBJECTIVE: Ibandronate is a highly potent nitrogen-containing bisphosphonate that can prevent bone loss in various animal models as well as in clinical trials. We evaluated the effects of different doses and treatment schedules in ovariectomized aged rats, a model of human osteoporosis. METHODS: Eight-month-old female Wistar rats were ovariectomized or sham operated (n = 15/group). Doses of 0.1 to 30 microg/kg/day ibandronate were administered subcutaneously over a period of 20 weeks with or without a 5 times higher single loading dose starting one day postsurgery. In a subsequent experiment, the optimal preventive dose, which is the lowest dose that prevented bone loss completely, or a suboptimal preventive dose were administered over the same period, either daily or by 3 cyclical intermittent regimens (on/off weeks = 1/2, 1/4, and 1/6), resulting in the same cumulative total dose. RESULTS: Ovariectomy induced significant bone loss in the following primary endpoints: femoral radiographic density, dry weight/tissue volume, and calcium content/tissue volume. Histomorphometry in the tibia resulted in reduced trabecular bone mass, thickness, and number, and increased separation. The optimal dose was 1.0 microg/kg/day, while 0.1 microg/kg/day was suboptimal. Higher doses resulted in a plateau. The loading dose had no effect on the results. Cyclical intermittent administration dose-dependently prevented bone loss, providing equivalent results per total dose, irrespective of the administration schedule. CONCLUSION: There were no differences between the various regimens, suggesting that it is the total dose of ibandronate rather than the treatment schedule that is important for efficacy, at least within the tested dosing intervals.     

Melatonin increases oestradiol-induced bone formation in ovariectomized rats

To assess the effect of melatonin on bone metabolism in ovariectomized rats, receiving oestradiol therapy or not, melatonin was administered in the drinking water (25 microg/mL water) and oestradiol (10 microg/kg body weight) or vehicle was given subcutaneously 5 days/week for up to 60 days after surgery. Urinary deoxypyridinoline (a marker of bone resorption) and circulating levels of bone alkaline phosphatase activity (a marker of bone formation), as well as serum calcium and phosphorus levels, were measured every 15 days. Bone area (BA), bone mineral content (BMC), bone mineral density (BMD) and total body fat (expressed as 100 g body weight) were measured by dual-energy X-ray absorptiometry at the end of the experiment. Body weight and total body fat were augmented after ovariectomy, and decreased after melatonin or oestradiol treatment. The effect of melatonin on body weight was seen in sham-operated rats only. Ovariectomy augmented, and melatonin or oestradiol lowered, urinary deoxypyridinoline excretion. This effect of melatonin and oestradiol was seen mainly in ovariectomized rats. The efficacy of oestradiol to counteract ovariectomy-induced bone resorption was increased by melatonin. Melatonin or oestradiol lowered serum bone alkaline phosphatase activity. Melatonin inhibition was seen mainly on the increase of bone alkaline phosphatase activity that followed ovariectomy. Serum phosphorus levels decreased after melatonin administration and were augmented after oestradiol injection; overall, melatonin impaired the increase of serum phosphorus caused by oestradiol. Ovariectomy decreased, and oestradiol increased, serum calcium levels while melatonin augmented serum calcium in sham-operated rats only. On day 60 after surgery, BMD and content decreased after ovariectomy and were increased after oestradiol injection. Melatonin augmented BA of spine and BMC of whole of the skeleton and tibia. The highest values observed were those of rats treated concurrently with oestradiol and melatonin. The present results indicate that: (i) melatonin treatment restrained bone remodelling after ovariectomy; (ii) the effect of melatonin required adequate concentrations of oestradiol; (iii) melatonin augmented oestradiol effects on bone in ovariectomized rats; (iv) a counter-regulation by melatonin of the increase in body fat caused by ovariectomy was uncovered. The melatonin doses employed were pharmacological in terms of circulating melatonin levels but not necessarily for some other fluids or tissues.     

铁调素对去卵巢大鼠骨代谢的影响

目的观察铁调素对去卵巢大鼠骨代谢的影响。方法将30只10周龄雌性Wistar大鼠分为假手术(sham)组、铁调素(hepc)组和生理盐水对照(OVX)组,每组10只。Hepc和OVX组行双侧卵巢摘除术后,分别经腹腔注射750μg/kg铁调素(hepcidin)溶液和等量生理盐水。9周后经颈静脉取血、处死后留股骨。检测股骨骨密度(BMD)、骨生物力学、血清骨代谢标志物及血清铁水平。结果与sham组相比,OVX组BMD及骨最大载荷、弯曲应力、弹性载荷、弹性模量均降低,差异均有统计学意义(P0.05),同时血清骨形成蛋白-2水平下降,而骨吸收指标Ⅰ型胶原交联C端肽(CTX-1)水平上升,血清铁浓度升高,差异均有统计学意义(P0.05);经铁调素干预的hepc组骨密度较OVX组增加了21.7%,上述骨生物力学指标也有明显改善,差异均有统计学意义(P0.05),虽然血清骨形成蛋白-2水平无明显升高,差异无统计学意义,但血清CTX-1水平下降33.5%,血清铁浓度降低17.2%,差异均有统计学意义(P0.05)。结论铁调素能改善绝经后骨质疏松大鼠模型骨代谢和力学特性,该作用可能通过改善体内铁超载、降低骨吸收实现。     

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.