补肾中药复方改善骨质疏松模型大鼠皮质骨生物力学性能的实验

背景：骨的生物力学性能优劣，取决于骨的结构和骨的材料。片面追求骨矿密度的提高，可能会引起骨生物力学性能的恶化。目前对骨质疏松治疗作用的不少研究过分强调了药物对骨矿密度的影响，对骨质疏松部位骨的整体生物力学性能的变化及其机制的研究还有待深入。设计：以实验动物为研究对象的完全随机对照的实验研究。目的：探讨补肾中药复方对去卵巢骨质疏松模型大鼠皮质骨(股骨)生物力学性能的作用及其相关的机制。单位：河南省洛阳正骨研究所生物医学工程实验室。材料：实验于2000-11／2001-06在河南省洛阳正骨研究所生物医学工程实验室完成。10月龄Wistar雌性大鼠50只，体质量(350&;#177;20)g。方法：将50只10月龄Wistar雌性大鼠随机分为5组：正常对照组、模型对照组、密骨胶囊组、仙灵骨葆组和倍美力组，每组10只。正常对照组做假手术，其余4组做卵巢切除术。术后正常饲养90d，第91天开始给药，连续用药90d，处死，测定股骨的骨矿密度、几何尺寸和股骨的生物力学性能。主要观察指标：①主要结局为股骨力学性能参数的比较。②次要结局为股骨几何结构参数、皮质骨面积和股骨各平面骨矿密度的变化。结果：模型大鼠股骨的生物力学性能明显下降，骨强度降低；股骨干外径变细，股骨中段骨皮质面积减少，骨矿密度有所下降。补肾中药复方组(密骨胶囊组和仙灵骨葆组)大鼠股骨的生物力学性能明显提高，骨强度增加；股骨中段外径增粗，骨皮质面积增加，骨矿密度提高。结论：补肾中药复方能明显改善去卵巢骨质疏松模型大鼠皮质骨(股骨)的生物力学性能。其主要作用机制是：补肾中药复方能够提高大鼠皮质骨(股骨)宏观结构的生物力学应答调节机能，使股骨干外径增粗，皮质骨面积增加，骨矿密度有所提高。     

补肾中药复方改善去卵巢骨质疏松模型大鼠松质骨骨生物力学性能的作用及机制探讨

目的探讨补肾中药复方改善去卵巢骨质疏松模型大鼠松质骨(腰椎)生物力学性能的作用及其相关的机制.方法将50只10月龄Wistar雌性大鼠随机分为正常对照组、模型对照组、密骨胶囊组、仙灵骨葆组和倍美力组.正常对照组做假手术,其余4组做卵巢切除术.术后正常饲养90d,第91天开始给药,连续用药90d,处死,电镜观察腰椎骨小梁的形态结构,测定腰椎骨质密度、骨质中钙、磷、有机质含量和生物力学性能.结果用药组大鼠股骨的生物力学性能明显提高,骨强度增加;骨质密度明显提高,骨质中钙、磷含量增加,骨有机质含量向正常组靠近,骨小梁的空间立体结构的破坏得到修复.结论补肾中药复方能明显改善去卵巢骨质疏松模型大鼠松质骨(腰椎)的生物力学性能.其主要作用机制是补肾中药复方能够修复松质骨被破坏的微细的空间立体形态结构,增加骨质密度,提高钙、磷含量,纠正骨代谢紊乱,调整骨质成分构成比例的平衡.     

仙灵骨葆治疗骨质疏松大鼠:血清学及骨组织形态计量学评价

背景:目前对于骨质疏松的治疗多为抑制骨吸收,有研究报道仙灵骨葆可促进骨形成,但具体机制不清.目的:通过干预去卵巢大鼠骨质疏松动物模型,探讨仙灵骨葆对骨质疏松大鼠骨量、生物力学性能及骨代谢的影响.方法:将24只大鼠随机分为正常对照组、模型组和仙灵骨葆组.除正常对照组外,其他2组行卵巢切除.6周后仙灵骨葆组给予仙灵骨葆干预,模型组给予等量双蒸水.4周后各组留取尿液、血清检测血Ⅰ型原胶原氨基端延长肽、尿脱氧吡啶诺啉排泄率和Ⅰ型胶原氨基末端肽排泄率.取各组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,行骨组织形态计量学检测.结果与结论:与模型组比较,仙灵骨葆组血Ⅰ型原胶原氨基端延长肽、尿脱氧吡啶诺啉排泄率、尿Ⅰ型胶原氨基末端肽排泄率,破骨细胞数、骨吸收周长百分数显著降低(P < 0.05),而远端骨密度和骨小梁体积均显著增高(P < 0.05),结果证实,仙灵骨葆可抑制骨吸收,促进骨形成,降低去卵巢大鼠骨转换水平,进而部分阻止其骨量丢失.     

补肾健骨中药对去势后大鼠骨质疏松症激素水平表达和骨密度影响的研究

目的:探究对去势后大鼠应用补肾健骨中药治疗骨质疏松症的疗效与作用机制。方法:研究用大鼠为Wister普通级大鼠,造模方法为将大鼠进行去卵巢处理来复制大鼠绝经后骨质疏松的实验模型,对实验大鼠进行随机分组:补肾健骨中药组、假手术组、雌二醇组、仙灵骨葆胶囊组、模型空白五组。补肾健骨中成药:仙灵骨葆胶囊。补肾健骨中药方剂治疗15周进行预防,阳性对照组使用尼尔雌醇、仙灵骨葆胶囊进行治疗,实验对照组设为模型空白对照组、假手术组。大鼠血清中促黄体生成素(LH)、骨钙素(BGP)、促卵泡激素(FSH)、雌激素(E2)水平检测运用放免方法检测;并检测大鼠骨密度(BMD),应用双能X线骨密度仪并取大鼠股骨与胫骨部位检测。结果:实验大鼠血清学指标结果比较:治疗后与空白对照组进行比较,补肾健骨中药组的E2的水平显著提高(P0.05),而BGP水平显著下降(P0.05),而激素FSH、LH水平均下降。实验大鼠经治疗后骨密度结果比较:治疗后与空白对照组进行比较,补肾健骨中药组变化差异最大(P0.05)。结论:补肾健骨中药治疗绝经后骨质疏松效果明显,可有效预防绝经后骨质疏松的发生。     

仙灵骨葆对大鼠骨质疏松性骨折愈合的影响

背景：骨质疏松可延迟骨折愈合,仙灵骨葆对骨质疏松性骨折愈合的影响及其机制尚有待研究。目的：观察骨质疏松对大鼠股骨干骨折愈合的影响以及仙灵骨葆的干预效果及其作用机制。方法：50只3月龄雌性SD大鼠随机分成5组：正常对照组、骨质疏松组、正常骨折组、骨质疏松性骨折组、治疗组,前2组每组6只,后3组每组12只。去除大鼠双侧卵巢制备骨质疏松模型,8周后制备右侧股骨干中段横行骨折模型。治疗组给予仙灵骨葆灌胃250mg/（kg·d）,正常骨折组、骨质疏松性骨折组灌胃等量生理盐水,灌胃6周。结果与结论：去卵巢后8周骨质疏松组的骨密度显著低于正常对照组（P〈0.05）,说明骨质疏松模型制作成功;灌胃6周,骨质疏松性骨折组骨密度、X射线摄片评分、最大载荷均显著低于正常骨折组和治疗组（P〈0.05）,但给予仙灵骨葆的治疗组仍显著低于正常骨折组（P〈0.05）。以上结果提示骨质疏松大鼠骨折愈合过程延迟,仙灵骨葆对骨质疏松大鼠骨折愈合具有积极促进作用。     

仙灵骨葆对骨质疏松性骨折骨痂血管形成的影响

背景：骨质疏松对骨折愈合早期的影响尚存争议,仙灵骨葆对骨质疏松性骨折愈合的影响及其机制尚有待深入研究。目的：观察骨质疏松对大鼠股骨干骨折愈合的影响,以及仙灵骨葆对骨质疏松性骨折愈合的作用。方法：将50只雌性12周龄Sprague-Dawley大鼠随机分成5组：假手术组、骨折组、去卵巢组、去卵巢＋骨折组及治疗组,后3组切除大鼠双侧卵巢制备去卵巢模型,骨折模型于去卵巢后4周制备,为股骨干中段横行骨折。治疗组在去卵巢及骨折基础上灌胃给予仙灵骨葆250mg/（kg·d）。给药3周,取去卵巢＋骨折组、骨折组和治疗组大鼠骨折侧标本行X射线摄像仪摄片后,测量其骨密度,苏木精-伊红染色观察骨痂组织的病理学改变并计数血管,免疫组织化学染色检测骨痂组织骨形态发生蛋白2的表达。结果与结论：去卵巢处理后大鼠的骨密度、计算机X射线摄像仪摄片评分显著降低（P〈0.05）,仙灵骨葆可在一定程度上提高去卵巢后骨折大鼠的骨密度及X射线摄像仪摄片评分,但差异无显著性意义（P〉0.05）;仙灵骨葆可提高去卵巢后骨折大鼠骨痂组织的血管数量（P〈0.05）,但对骨形态发生蛋白2的表达无影响。提示,去卵巢后大鼠骨折早期愈合过程延迟,仙灵骨葆可促进骨质疏松大鼠骨折愈合早期血管的形成。     

仙灵骨葆对骨质疏松性骨折骨痂血管形成的影响

背景:骨质疏松对骨折愈合早期的影响尚存争议,仙灵骨葆对骨质疏松性骨折愈合的影响及其机制尚有待深入研究。目的:观察骨质疏松对大鼠股骨干骨折愈合的影响,以及仙灵骨葆对骨质疏松性骨折愈合的作用。方法:将50只雌性12周龄Sprague-Dawley大鼠随机分成5组:假手术组、骨折组、去卵巢组、去卵巢+骨折组及治疗组,后3组切除大鼠双侧卵巢制备去卵巢模型,骨折模型于去卵巢后4周制备,为股骨干中段横行骨折。治疗组在去卵巢及骨折基础上灌胃给予仙灵骨葆250mg/(kg·d)。给药3周,取去卵巢+骨折组、骨折组和治疗组大鼠骨折侧标本行X射线摄像仪摄片后,测量其骨密度,苏木精-伊红染色观察骨痂组织的病理学改变并计数血管,免疫组织化学染色检测骨痂组织骨形态发生蛋白2的表达。结果与结论:去卵巢处理后大鼠的骨密度、计算机X射线摄像仪摄片评分显著降低(P<0.05),仙灵骨葆可在一定程度上提高去卵巢后骨折大鼠的骨密度及X射线摄像仪摄片评分,但差异无显著性意义(P>0.05);仙灵骨葆可提高去卵巢后骨折大鼠骨痂组织的血管数量(P<0.05),但对骨形态发生蛋白2的表达无影响。提示,去卵巢后大鼠骨折早期愈合过程延迟,仙灵骨葆可促进骨质疏松大鼠骨折愈合早期血管的形成。     

仙灵骨葆治疗骨质疏松大鼠：血清学及骨组织形态计量学评价

背景：目前对于骨质疏松的治疗多为抑制骨吸收,有研究报道仙灵骨葆可促进骨形成,但具体机制不清。目的：通过干预去卵巢大鼠骨质疏松动物模型,探讨仙灵骨葆对骨质疏松大鼠骨量、生物力学性能及骨代谢的影响。方法：将24只大鼠随机分为正常对照组、模型组和仙灵骨葆组。除正常对照组外,其他2组行卵巢切除。6周后仙灵骨葆组给予仙灵骨葆干预,模型组给予等量双蒸水。4周后各组留取尿液、血清检测血Ⅰ型原胶原氨基端延长肽、尿脱氧吡啶诺啉排泄率和Ⅰ型胶原氨基末端肽排泄率。取各组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,行骨组织形态计量学检测。结果与结论：与模型组比较,仙灵骨葆组血Ⅰ型原胶原氨基端延长肽、尿脱氧吡啶诺啉排泄率、尿Ⅰ型胶原氨基末端肽排泄率,破骨细胞数、骨吸收周长百分数显著降低（P〈0.05）,而远端骨密度和骨小梁体积均显著增高（P〈0.05）,结果证实,仙灵骨葆可抑制骨吸收,促进骨形成,降低去卵巢大鼠骨转换水平,进而部分阻止其骨量丢失。     

仙灵骨葆对尾悬吊拟失重大鼠骨量丢失的抑制

背景:以淫羊藿苷为主要成分的中药制剂仙灵骨葆具有促进骨形成的作用。目的:观察仙灵骨葆对尾悬吊拟失重大鼠骨量丢失的抑制作用及机制。方法:将SD大鼠随机分为正常对照组、拟失重组、仙灵骨葆组,后两组大鼠采用尾悬吊法模拟失重,仙灵骨葆组悬吊同时灌胃给予仙灵骨葆250mg/(kgd),拟失重组悬吊同时灌胃给予双蒸水,4周后取右侧股骨行骨密度、三点弯曲生物力学及骨组织形态计量学检测。结果与结论:正常对照组骨密度、胫骨近端骨小梁相对体积、骨小梁厚度、骨小梁数量、生物力学最大载荷显著高于拟失重组、仙灵骨葆组(P<0.05)。仙灵骨葆组胫骨远端1/4骨密度、胫骨近端骨小梁相对体积显著高于拟失重组(P<0.05),生物力学最大载荷也高于拟失重组,但差异无显著性意义。说明仙灵骨葆灌胃干预可促进松质骨的骨形成,部分阻止拟失重大鼠骨量的丢失。     

仙灵骨葆对骨质疏松大鼠骨量及骨髓基质干细胞成骨分化的影响

背景：双侧卵巢切除可造成大鼠骨量丢失,仙灵骨葆作为传统中药具有一定的促进骨形成作用。目的：观察仙灵骨葆体内给药对骨质疏松火鼠骨量及骨髓摧质干细胞成骨分化能力以及相关因子表达的影响。方法：3月龄雌性SD大鼠24只随机数字表法均分为3组,卵巢切除组和仙灵骨葆组行卵巢切除,造模6周后仙灵骨葆组给予仙灵骨葆250mg／（kg·d）,干预8剧：正常对照组不予干预。结果与结论：卵巢切除组L2-L4椎体骨密度妊著低于其他两组,仙灵骨葆组仍显著低于正常对照组（P〈0．05）;卵巢切除组血清骨钙素、骨髓基质干细胞的骨形态发生蛋白2、骨钙素mRNA水平均低于其他两组（Pc0．05）。细胞外基质矿化能力亦明履低于正常对照组和仙灵骨葆组。提示大鼠去势14周后骨量丢失显著,仙灵骨葆可部分阻止其骨量丢失,其作用机制可能与促进大鼠骨髓綦质干细胞的成骨分化有关。     

他莫西芬对去势大鼠骨生物力学性能的影响

目的评价他莫西芬对雌激素缺乏引起的骨生物力学性能下降的预防作用。方法采用4月龄Sprague-Dawley(SD)雌性大鼠切除卵巢作动物模型,术后给予他莫西芬治疗连续12周。结果与假手术组相比去势组骨密度、股骨生物力学性质、腰椎骨形态计量指标均有显著差异;他莫西芬处理组与去势组比较,股骨骨密度、股骨最大载荷、桡度、最大应力、弹性模量显著升高,骨形态计量参数的改变均有显著性。结论他莫西芬能抑制去势引起的SD大鼠骨生物力学性能下降。     

复方脑复康对去卵巢骨质疏松大鼠股骨骨密度及生物力学的影响

目的探讨复方脑复康对去卵巢(OVX)骨质疏松大鼠股骨骨密度及股骨生物力学的影响。方法40只6、7月龄雌性SD大鼠随机分为4组:假手术(Sham)组、OVX组、OVX+己烯雌酚(E)组和OVX+复方脑复康(F)组,每组10只。实行手术后,于术后第3天开始灌胃用药。90天后处死大鼠,取左侧股骨进行股骨骨密度和生物力学的测量。结果与Sham组比,OVX组股骨骨密度及生物力学指标明显下降,全股骨密度、弹性载荷和弯曲能量差异有显著性(P<0.05),而用己烯雌酚和复方脑复康能阻止这种变化;与E组相比,F组的这种阻止作用更为明显。结论复方脑复康能防治OVX造成的骨质疏松,增加骨密度,防止骨生物力学性能的受损,效果优于单独应用己烯雌酚治疗。     

Dose effects of propranolol on cancellous and cortical bone in ovariectomized adult rats

Animal studies suggest that bone remodeling is under beta-adrenergic control via the sympathetic nervous system. The purpose of this study was to examine the preventive effect of different doses of nonspecific beta-blockers (propranolol) on trabecular and cortical bone envelopes in ovariectomized rats. Six-month-old female Wistar rats were ovariectomized (OVX, n = 60) or sham-operated (n = 15). Then, OVX rats were subcutaneously injected with 0.1 (n = 15), 5 (n = 15), or 20 (n = 15) mg/kg propranolol or vehicle (n = 15) for 10 weeks. Tibial and femoral bone mineral density (BMD) were analyzed longitudinally by dual-energy X-ray absorptiometry. At death, the left tibial metaphysis and L(4) vertebrae were removed, and microcomputed tomography (Skyscan 1072; Skyscan, Aartselaar, Belgium) was performed for trabecular bone structure investigation. Histomorphometry analysis was performed on the right proximal tibia to assess bone cell activities. After 10 weeks, OVX rats had decreased BMD and trabecular parameters and increased bone turnover, as well as cortical porosity compared with the sham group (p < 0.001). Bone architecture alteration was preserved by 0.1 mg/kg propranolol due to higher trabecular number and thickness (+50.35 and +6.81%, respectively, than OVX; p < 0.001) and lower cortical pore number (-52.38% than OVX; p < 0.001). Animals treated by 0.1 mg/kg propranolol had a lower osteoclast surface and a higher osteoblast activity compared with OVX. Animals treated by 20 mg of propranolol did not significantly differ from OVX rats. Animals treated by 5 mg of propranolol have been partially preserved from the ovariectomy. These results showed a dose effect of beta-blockers. The lower the dose of propranolol breeding, the better the preventive effect against ovariectomy.     

Regional bone mineral in healthy and osteoporotic women: a cross-sectional study

Regional bone mineral content and density (BMC and BMD) was measured in six regions (head, arms, chest, spine, pelvis, and legs) using dual photon 153Gd absorptiometry (DPA) in 128 healthy women aged 21-77 years, and in 45 women presenting with Colles' fracture (mean age 65 years), 46 women with vertebral crush or wedge fracture (mean age 68 years), and 27 women with femoral neck-fracture (mean age 74 years). The age-related normal bone loss was generalized, uniformly distributed, and best described by a combination of a premenopausal linear and a postmenopausal exponential regression in all six regions. Looking at BMD, the overall expected bone loss from age 20 to age 80 was approximately 20% in all the regions. When the fracture patients were examined, we found also generalized bone deficit as the prominent feature, amounting to about 20% of the premenopausal level for Colles' and spinal fractures, and about 25% for femoral neck-fracture. However, there was a regional bias in the fracture patients, as the Colles' and spinal fracture patients had a preferential reduction in spinal and pelvic BMD, whereas the patients with femoral neck-fracture had a preferential reduction in pelvic and leg BMD. We conclude that age-related and osteoporotic bone loss is generalized. Furthermore, we propose that regional differences in osteoporotic bone loss are brought about by a simple biological variability of the range of (i) relative amount of trabecular and cortical bone, (ii) rate of loss in the two types of bone tissue, and (iii) time of onset of trabecular relative to cortical bone loss.     

卵巢切除股骨骨折大鼠骨愈合中降钙素的作用

背景：降钙素可活化腺苷酸环化酶蛋白激酶A通路及磷脂酶C通路,抑制破骨细胞的活性,可能治疗骨质疏松性骨折。目的：观察降钙素对去卵巢大鼠股骨骨折愈合的作用。方法：构建双侧卵巢切除骨质疏松右股骨骨折SD大鼠模型,然后分别皮下注射生理盐水和降钙素（16IU/kg）,隔日1次,于骨折后3周和6周测量右股骨行骨密度,苏木精-伊红及抗酒石酸酸性磷酸酶染色,骨形态发生蛋白2及血管内皮生长因子免疫组化染色。结果与结论：骨折后给予降钙素治疗的大鼠抗酒石酸酸性磷酸酶染色阳性细胞积分吸光度值较生理盐水治疗的大鼠显著减少（P〈0.05）。骨折后3周,两组骨折线均较清晰,骨痂体积无明显差别,骨折愈合以软骨内化骨过程为主,骨密度无显著性差异（P〉0.05）。骨折后6周,两组骨折线较模糊,骨痂体积无差别,骨小梁排列较有序,用药组股骨骨密度较对照组升高（P〈0.05）。两组在骨折后3周和6周的骨形态发生蛋白2及血管内皮生长因子差异无显著性意义（P〉0.05）。证实降钙素可以抑制去卵巢大鼠骨折部位破骨细胞活性,但无明显促进大鼠股骨骨折愈合的作用。     

绝经后女性股骨颈容积性定量CT测量研究

目的应用vQCT技术测量绝经后女性股骨颈的容积性骨密度（BMD）和几何参数,评价各参数对骨质疏松性骨折的预测能力。方法选取绝经后妇女47例,其中正常组26例,骨质疏松组14例,骨质疏松伴腰椎骨折组7例。应用多层螺旋CT对三组患者左股骨颈进行容积扫描并薄层重建,利用Osteo CAD软件对多平面重组图像进行测量,计算出股骨颈的皮质骨、小梁骨和整体骨的容积BMD,股骨颈轴长和最小横截面积,并进行统计学分析。结果正常组与骨质疏松组、正常组与骨折组之间vQCT及DXA各BMD差异均有统计学意义,在调节年龄、身高和体重因素后差异仍存在;骨质疏松组与骨折组之间只有小梁BMD及总体BMD差异有统计学意义,且小梁BMD下降幅度较大,达32.4%。结论股骨近端vQCT测量比DXA能更敏感地反映绝经后女性股骨颈BMD的变化情况,为早期预测骨质疏松性骨折提供科学依据。     

补肾健脾方干预去势大鼠骨骼肌caspase-3和caspase-8的表达

背景：骨骼肌与骨质疏松症关系密切,导致其衰退的主要原因是细胞的凋亡,这种凋亡可能是Caspase家族蛋白所介导的。目的：探讨补肾健脾方对去势大鼠骨骼肌caspase-3和caspase-8调控作用。方法：SD大鼠48只等随机分为对照组、模型组、中药组、西药组,后3组大鼠摘除双侧卵巢建立骨质疏松模型,对照组仅切除周围少量脂肪组织。造模2周后,中药组给予补肾健脾方（2.979g/kg）灌胃,西药组给予戊酸雌二醇片（0.104mg/kg）灌胃,模型组和对照组给予蒸馏水灌胃,1次/d。结果与结论：①干预12周后,双能X线骨密度仪测量发现,相比于对照组,模型组大鼠左侧股骨的骨密度值和骨矿含量均明显降低（P〈0.01）;而中药组和西药组的骨密度值均高于模型组（P〈0.05）,且2组骨密度值和骨矿含量接近（P〉0.05）。②酶标免疫分析显示,与对照组相比,模型组骨骼肌中Caspase-3和Caspase-8表达水平明显上升（P〈0.05）。中药能显著降低大鼠骨骼肌中Caspase-3的表达水平（P〈0.05）,而西药能显著减低Caspase-8表达水平（P〈0.05）。③说明补肾健脾方对卵巢切除所致的骨质疏松症有明显的治疗作用,能明显提高骨密度,而且能显著减低Caspase-3的水平,但不能显著减低Caspase-8水平,显示补肾健脾方不是通过死亡受体途径去抑制细胞凋亡的。     

Polymorphisms in the vitamin D receptor gene and bone mass, bone turnover and osteoporotic fractures

BACKGROUND: Vitamin D is essential for normal bone metabolism. Polymorphisms in exon 2, intron 8 and exon 9 of the vitamin D receptor (VDR) gene have previously been found to be associated with bone mass and bone turnover. MATERIALS AND METHODS: We examined the effect of these polymorphisms, separately and in combination, on bone mineral density (BMD), bone turnover, and the prevalence of osteoporotic fractures in 192 osteoporotic patients and 207 normal controls. The four polymorphisms were determined by RFLP using Fok I (T2-C), Bsm I (intron 8), Apa I (intron 8) and Taq I (T1055-C) after PCR. RESULTS: We did not find any association between the Fok I polymorphism and bone mass, bone turnover or prevalence of osteoporotic fractures. We found that BB + Bb-genotypes were more frequent in patients with osteoporotic fractures (chi2 = 3.50, P = 0. 06). Furthermore, BMD of the intertrochanteric region (P < 0.0001, ANOVA) as well as the total hip (P < 0.01, ANOVA) were higher in individuals with the bb-genotype. The Apa I and the Taq I polymorphisms were not distributed differently among osteoporotic patients and normal controls. Apa I was not associated with differences in BMD. BMD of the intertrochanteric region was higher in individuals with the TT-genotype compared with individuals with the Tt- or tt-genotypes (P < 0.01, ANOVA), while no differences could be demonstrated in BMD of the lumbar spine, femoral neck, trochanter or Wards triangle. Combining the genotypes generally reflected the differences caused by the Bsm I polymorphism. CONCLUSION: We have found that the B-allele of the Bsm I polymorphism in the 3' untranslated region of the VDR was associated with low BMD at the hip, and tended to be associated with osteoporotic fractures. The translation initiation polymorphism in the VDR does not affect BMD and is not associated with osteoporotic fractures in men or women.     

Synthesis, characterization and evaluation of bone targeting salmon calcitonin analogs in normal and osteoporotic rats

In order to assess the therapeutic efficacy of an antiresorptive drug with imparted bone targeting potential using bisphosphonate (BP) conjugation and an improved pharmacokinetic profile using PEGylation, we synthesized, characterized and evaluated in vivo efficacy of bone-targeting PEGylated salmon calcitonin (sCT) analog (sCT-PEG-BP). sCT-PEG-BP was compared with non-PEGylated bone targeting sCT analog (sCT-BP) and unmodified, commercially available sCT. sCT-PEG-BP conjugates were characterized by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) analysis. The effect of PEG-BP or BP upon sCT secondary structure was examined by Circular Dichroism and sCT-PEG-BP was evaluated for in vitro bone mineral Hydroxyapatite (HA) binding ability and calcium salts specificity using a binding assay for bone HA and several calcium salts. Anti-calcitonin antibody binding ability of these analogs was determined using enzyme-linked immunosorbent assay (ELISA), by reacting bone targeting sCT analogs with calcium phosphate coated Osteologic? plates and detecting the bound sCT using anti-sCT antibody. Potential cytotoxicity of these compounds was evaluated in monocytic RAW 264.7 cells, and sCT bioactivity was evaluated using an in vitro intracellular cAMP stimulation assay in human T47D breast cancer cells. Finally, in vivo efficacy of each compound was evaluated by determining the plasma levels of calcium after s.c. administration in normal rats, and in a rat model of Osteoporosis, secondary to ovariectomy (OVX). In vivo micro-computed tomography (micro-CT) was used to temporally map and quantify alterations in bone volume and bone mineral density (BMD) in the same animals at 1, 4, 8 and 12 weeks after OVX surgery. Sixteen 6 week old virgin female rats underwent OVX surgery followed by the daily s.c. injection of 2.5IU/kg/day sCT or equivalent analogs, and compared to four sham-operated, placebo treated control rats. Our results showed the chemical coupling of PEG-BP or BP to sCT altered its secondary structure without altering its antibody binding ability. sCT analogs retained strong sCT bioactivity, were non-toxic to RAW 264.7 cells in culture and elicited a comparable hypocalcemic effect to that of unmodified sCT in normal rats. Compared to marketed unmodified sCT, sCT-PEG-BP showed significantly improved efficacy in terms of preserving bone volume, BMD and trabecular micro-architecture in osteoporotic rats at the initial dose tested. Bisphosphonate-mediated targeting of PEGylated sCT to bone represents a new class of targeted antiresorptive compounds that has not previously been attempted.     

复合振动对卵巢切除大鼠骨质量的作用

背景:目前所使用的全身振动防治骨质疏松所需振动强度较大,人体不适感较强.作者设计了复合振动,前期实验发现复合振动可在更低强度下有效预防卵巢切除大鼠的骨密度下降. 目的:课题创新性提出低强度复合振动可维持生长期卵巢切除SD大鼠骨质量的理论假设,并期望实验结果加以验证.方法:SPF级4月龄雌性未育SD大鼠32只,随机分为正常对照组、卵巢切除组以及振动1、振动2组,每组大鼠均为8只.振动1组接振45～55Hz,0.05～0.1 g;振动2组接振45～55Hz,0.12～0.21 g.振动20min/次,1次/d,5次/周,休息间隔不大于2d.实验时间13周.观察振动干预前后大鼠活体骨密度,体外标本骨微结构以及生物力学性能. 结果与结论:卵巢切除组腰椎骨密度下降(P<0.05),而正常对照组与两振动组有显著性增加,股骨骨密度均增加,组间差异无显著性意义;卵巢切除各组骨微结构参数均明显下降,但振动2组骨小梁数量、骨小梁厚度、骨小梁间距、骨体积分数相对于卵巢切除组有显著改善:腰椎骨强度值两振动组较卵巢切除组显著增加(P=0.025、0.006),与正常对照组比较差异无显著性意义.实验结果证明,特定的复合振动舒适感较好的低强度下可以有效预防卵巢切除SD大鼠骨密度下降,减轻骨微结构破坏程度,维持骨强度,具有改善卵巢切除大鼠骨质量的作用和潜在的预防骨质疏松作用.