仙灵骨葆改善骨质疏松大鼠股骨骨生物力学性能的作用及机制研究

目的探讨补肾中药仙灵骨葆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。方法将40只10月龄wistar雌性大鼠随机分为仙灵骨葆组、倍美力组、正常组和模型组,每组10只。正常组做假手术,其余3组做卵巢切除术。术后正常饲养90 d。第91天开始,仙灵骨葆组、倍美力组分别给予临床等效剂量相应药物:仙灵骨葆55 mg/d(胶囊量),倍美力0.01 mg/d,正常组和模型组不给药。连续给药90 d后处死大鼠,测定股骨的几何尺寸、极惯性矩、抗扭截面模数、截面惯性矩、抗弯截面模数、骨矿密度和股骨的生物力学试验。结果模型大鼠股骨干外径变细,骨皮质面积减少,抗弯、抗扭截面模数变小,骨矿密度减损;股骨的生物力学性能明显下降,骨强度降低。用药组大鼠股骨干外径增粗,骨皮质面积增加,抗弯、抗扭截面模数增大,骨矿密度提高。股骨的生物力学性能明显改善,骨强度增加。结论仙灵骨葆能够提高股骨宏观结构的生物力学应答调节功能,使股骨干外径增粗,皮质骨面积增加,骨丢失减少使股骨结构的抗弯、抗扭截面模数增大,使骨的力学结构优化,骨强度提高。     

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.     

补肾中药组方对去卵巢大鼠骨生物力学的影响

目的 研究补肾中药组方（CKD）对去卵巢大鼠骨生物力学的影响.方法 60只雌性大鼠随机分为假手术组、卵巢切除（OVX）模型组、OVX＋CKD高剂量组、OVX＋CKD中剂量组、OVX＋CKD低剂量组和OVX＋雌二醇（E2）组,每组10只.3个月后,采用双能X线骨密度测定仪测定骨矿物质密度（BMD）并比较不同剂量CKD对OVX大鼠骨生物力学的影响.结果 模型组大鼠BMD有明显的下降（与假手术组比较,P＜0.05）;与模型组比较,小、高、中剂量组骨的最大载荷和挠度提高（P＜0.05）,高、中剂量组骨的弹性模量降低（与模型组比较,P＜0.05）,这种作用与尼尔雌醇作用相当（P＞0.05）;且补肾中药组方无明显的副作用.结论 补肾中药具有改善OVX大鼠骨生物力学的作用.     

补肾健骨汤对去卵巢大鼠骨质疏松症骨密度和骨生物力学性能影响的实验研究

目的观察补肾健骨汤对去卵巢大鼠骨质疏松症骨密度和骨生物力学性能的影响。方法将90只6月龄雌性Wistar大鼠[体重(300±20)g]随机分为空白对照组15只,造模组(OVX)75只。造模组造模成功后再将其随机分为5组,即模型对照组15只,补肾健骨汤高剂量组15只,补肾健骨汤中剂量组15只,补肾健骨汤低剂量组15只,雌激素组15只。复制骨质疏松症动物模型,造模成功后,分组给药。给药3个月后所有动物统一处死取材。结果 (1)与空白对照组比较,模型对照组股骨骨密度值及骨矿物含量有明显下降(P0.05);与模型对照组相比,补肾健骨汤中高剂量组与雌激素组的骨密度及骨矿盐含量均有明显提高(P0.05),补肾健骨汤组高剂量组与雌激素组无明显差异。(2)补肾健骨汤高剂量组和模型对照组相比,大鼠股骨的弹性模量、最大载荷、屈服强度均有不同程度的改善,差异有统计学意义(P0.05),与空白对照组相比差异无统计学意义(P0.05)。结论去卵巢大鼠骨密度及骨矿物含量明显降低;补肾健骨汤能明显改善去卵巢大鼠骨密度及骨矿物含量和骨生物力学性能。     

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。     

强骨宝对去势大鼠骨形态计量学影响的研究

目的 观察强骨宝（黄芪＋司坦唑醇,JGB）对去卵巢大鼠骨质疏松症骨形态计量学参数的影响.方法 取32只Wistar雌性大鼠随机分为4组,行去势手术,制成去卵巢大鼠骨质疏松模型.然后分别灌胃给药生理盐水、己烯雌酚、强骨宝,8 w后处死,取大鼠胫骨上段松质骨和中段皮质骨硬组织包埋、切片,图像分析松质骨、皮质骨形态计量学参数.结果 强骨宝可使去卵巢大鼠松质骨静态参数和动态参数显著增加,吸收参数显著降低;皮质骨面积、骨髓腔面积等静态及内外骨膜骨形成动态参数均无显著变化,内膜面骨吸收参数降低.结论 强骨宝能有效预防去卵巢大鼠骨质疏松症松质骨丢失,亦具有抑制皮质骨内膜骨吸收的作用.     

补肾活血方对去势大鼠骨质疏松的影响

目的观察补肾活血方对去势大鼠的血生化、骨密度、生物力学及病理学的影响,评价补肾活血方防治去势大鼠骨质疏松的疗效。方法取36只雌性大鼠随机分为对照组、假手术组和补肾活血组。补肾活血组与对照组大鼠行卵巢切除术,假手术组行假手术处理,均饲养3个月后行病理检查确认造模成功。造模成功后各组大鼠分别给予0．9％氯化钠注射液、0．9％氯化钠注射液、补肾活血中药灌胃,3个月后处死取血分离得到血清,分别检测血清雌激素（E：）、碱性磷酸酶（ALP）和骨钙素（BGP）。取大鼠腰椎、股骨测量腰椎的骨密度（BMD）和最大承载力、行股骨病理学观察。结果与对照组比较,补肾活血方组大鼠血清E：升高,差异有统计学意义（P〈0．01）;骨形成指标ALP、BGP上升,差异有统计学意义（P〈0．05）。与对照组比较,补肾活血组大鼠腰椎BMD及最大承载力均升高,差异有统计学意义（P〈0．05）。HE染色病理组织形态观察发现补肾活血方组大鼠骨小梁稍细,无明显变薄,排列尚整齐并连接成网,密度、面积尚正常,部分区域骨小梁间隙略增大,较对照组有明显改善。结论补肾活血方能够提高去势大鼠雌激素水平,促进骨形成,增加骨量,提高骨组织的力学性能,改善骨组织状况,达到防治骨质疏松的疗效。     

补肾中药组方对去卵巢大鼠骨微结构的影响

目的研究补肾中药组方对去卵巢大鼠骨微结构的影响。方法通过卵巢摘除建立骨质疏松模型,研究不同剂量的补肾中药对去卵巢大鼠骨微结构的影响。结果光镜下观察高剂量补肾中药组方能改善去卵巢大鼠的骨小梁的分布、排列、之间的连接、厚度及间隙等组织学表现。组织计量学表明高、中剂量中药组能提高骨小梁TBV%和MTT(与OVX组比较P<0.05),这种功能与尼尔雌醇作用相当(P>0.05),低剂量中药组能提高骨小梁TBV%(与OVX组比较P<0.05)。结论补肾中药具有改善去势大鼠骨微结构的功能。     

不同月龄大鼠骨矿含量、骨生物力学改变及大豆苷元的干预作用

目的 探讨不同月龄大鼠骨生物力学的改变及补充大豆苷元对其变化的影响.方法 60只3月龄雌性大鼠,20只假手术,30只切除双侧卵巢,按体重随机分为6组:3月龄基础对照组(3-Yong),6月龄假手组(6-Sham),6月龄去卵巢组(6-OVX),9月龄假手术组(9-Sham ),9月龄去卵巢组(9-OVX),9月龄灌喂75 mg/kg大豆苷元组(9-OVX-D75).大鼠处死后,取下后肢,分离软组织,采用质构仪进行3点弯曲实验,测定左侧股骨生物力学指标最大荷载、最大挠度等,并测定骨矿含量.结果 大鼠去卵巢后,股骨生物力学特性及骨矿含量均降低,但随着月龄的增长,大鼠股骨承受最大载荷增加,骨矿含量也增加,给予大豆苷元后,可改善大鼠股骨生物力学特性.结论 大鼠去卵巢后不但可以导致骨量的丢失,而且使骨生物力学性能下降.补充大豆苷元可较好改善股骨的挠度和最大载荷,增强抗性.     

补骨制剂Ⅱ对摘除卵巢大鼠骨质疏松的防治作用

目的研究补骨制剂Ⅱ经灌胃给药后对摘除卵巢大鼠骨质疏松模型的影响。方法雌性6月龄SD大鼠分为假去势组,模型组,阳性对照组,补骨制剂Ⅱ高、中、低剂量组(n=12),除假去势组进行假手术外,其余均手术彻底摘除卵巢法,术后进行模型筛选,9 d后假去势组、去势组均给予生理盐水,阳性对照组灌胃给予己烯雌酚0.02 mg/kg、鱼肝油450 U/kg、多种钙片0.5 g/kg,补骨制剂Ⅱ高、中、低剂量组分别灌胃给予补骨制剂Ⅱ稠浸膏(剂量以生药量计)12、6、3 g生药/kg,连续给药3个月。末次给药后,检测血清钙(Ca2+)、血清磷(P),超氧化物歧化酶(SOD)、丙二醛(MDA)、血清雌二醇(E2)含量,并测定骨密度、骨生物力学和骨组织形态。结果与去势组相比,低、中、高剂量补骨制剂Ⅱ组在鼠血清中各项检测指标均有显著差异(P0.05),增加腰椎骨密度,提高模型大鼠的股骨最大载荷和刚度,改善骨小梁数量和连续性。结论补骨制剂Ⅱ可明显改善去卵巢大鼠的骨生物力学性能,提高去势大鼠血清中雌激素水平,能防治去卵巢诱导的大鼠骨质疏松,是值得开发的一种中药复方制剂。     

补肾益精方对切除卵巢大鼠骨质疏松模型腰椎骨生物力学性能的影响

目的：探讨被 肾益精方对去卵巢大鼠骨质疏松模型松质骨生物力学性能的影响,方法：40只10月龄Wistar雌性大鼠随机分为正常对照组,模型对照组,补肾益精方组和倍美力组,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天,处死,取出第3腰椎,测定其骨质含量并进行压应力测试与分析,结果：补肾益精方组比模型组腰椎骨的骨质含量提高了6．19％,最大承载力增加了21．66％,极限强度增加了16．93％,结论：应用补肾益精方可以明显改善去卵巢大鼠骨质疏松模型松质骨的生物力学性能。     

去卵巢大鼠血清乳铁蛋白与骨密度的相关性研究

目的 探讨去卵巢大鼠血清乳铁蛋白(LF)水平与骨密度(BMD)的关系,为临床应用乳铁蛋白提供实验依据.方法 采用4月龄Wistar雌性大鼠,随机分为假手术组(sham,20只)与去卵巢模型组(ovariectomy,OVX,40只,分为OVX I组与OVX Ⅱ组,各20只),分别于术后14与18周处死OVX I组与OV...     

叶酸对去卵巢大鼠骨质疏松的保护作用

目的 探讨叶酸对去卵巢大鼠骨质疏松的保护作用.方法 40只3月龄雌性SD大鼠随机分为5组:假手术组、去卵巢组、乙烯雌酚组(乙烯雌酚0.03 mg·kg-1·d-1)、低剂量叶酸组(叶酸5 mg·kg-1·d-1)、高剂量叶酸组(叶酸20 mg·kg-1·d-1).各组大鼠于术后1周开始灌胃给药,治疗10周,假手术组和去卵巢组给予溶媒灌胃.测定大鼠血浆总同型半胱氨酸(tHcy)浓度,骨匀浆中碱性磷酸酶(ALP)和抗酒石酸酸性磷酸酶(TRACP)的水平;取右股骨和腰5椎体进行骨密度和骨生物力学测定,取腰6椎体和左股骨制备HE切片,观察骨组织的形态学变化.结果 与假手术组比较,去卵巢大鼠血浆tHcy浓度明显升高,腰椎和股骨骨密度显著减低(均P＜0.01),血浆tHcy浓度与腰椎骨密度呈负相关(r=-0.359,P=0.040).叶酸显著降低去卵巢大鼠血浆tHcy浓度(均P＜0.01).大剂量叶酸显著增加去卵巢大鼠骨匀浆ALP水平,降低TRACP水平,增加腰椎和股骨骨密度(均P＜0.01),改善腰椎和股骨的生物力学性能.结论 去卵巢大鼠存在高同型半胱氨酸血症,高同型半胱氨酸参与了去卵巢大鼠骨质疏松的发生.叶酸对去卵巢大鼠骨质疏松具有保护作用,其机制可能与改善同型半胱氨酸的代谢作用有关.

Abstract：

Objective To investigate the protective effect of folic acid(FA) on osteoporosis in ovariectomized(OVX) rats.Methods Forty three-month-old female SD Rats were divided into 5 groups, sham operation group, OVX group, diethylstilbestrol group(0.03mg·kg-1·d-1),low dose FA Group (5 mg·kg-1·d-1),and high dose FA group (20 mg·kg-1·d-1).Gastric gavage in each group was started from one week after being ovariectomized and lasted 10 weeks. Sham operation group and OVX group were treated with solvent. The rats were sacrificed at the end of 10th week after treatment. The total homocysteine(tHcy) in plasma, alkaline phosphatase(ALP), and tartrate-resistant acid phosphatase(TRACP) activity of bone homogenates were measured. The bone mineral density(BMD) and bone biomechanics were determined using L5 vertebrae and right femur. The bone tissue slices were made with L6 vertebrae and left femur and HE stained, and then the histomorphology was observed. Results Compared with sham operation group, plasma tHcy level was significantly increased(P＜0.01), BMD of lumbar vertebrae and femur was remarkedly decreased in OVX group(all P＜0.01). Plasma tHcy concentration was negatively correlated with lumbar BMD(r=-0.359, P=0.040). Plasma tHcy level in both groups treated with folic acid was significantly reduced(all P＜0.01). The ALP concentration in bone homogenates was higher, the TRACP concentration in bone homogenates was lower, and BMD and bone biomechanics of lumbar vertebrae and femur were increased in high dose FA group than those in OVX group(all P ＜0.01). Conclusions In OVX rats hyperhomocysteinemia existed and was involved in the development of osteoporosis. Folic acid could protect OVX rats from osteoporosis, due probably to improved homocysteine metabolism.     

Increased bone volume and bone mineral content in xylitol-fed aged rats.

BACKGROUND: Our previous studies have shown that dietary xylitol supplementation protects against the loss of bone mineral after ovariectomy. The ovariectomy-induced decrease in trabecular bone volume is significantly retarded by dietary xylitol. Objective: To study whether dietary xylitol can protect against bone loss also during aging, a long-term experimental study was performed with rats. METHODS: Twenty-four male Sprague-Dawley rats were divided into two groups. The rats in the control group were fed a basal RM1 diet, while the rats in the experimental group were continuously fed the same diet supplemented with 10% (w/w) xylitol. The rats were killed after 20 months. Their tibiae were used for the analyses of bone density and trabecular bone volume, and their femurs were used for the scanning analyses with peripheral quantitative computed tomography (pQCT). RESULTS: The tibial density of the xylitol-fed aged group (1.73 +/- 0.14 g/mm(3)) was significantly greater than that of the aged group without xylitol (1.56 +/- 0.14 g/ mm(3)). The trabecular bone volume of the xylitol-fed rats was 21.2 +/- 4.0%. It was significantly greater than that of the rats not receiving xylitol (9.3 +/- 4.3%). The pQCT-measured cortical bone mineral density and the pQTC-measured cortical bone mineral content of the femoral diaphysis were significantly greater in the xylitol-fed group than in the control group. The trabecular bone mineral density and the trabecular bone mineral content of the femoral distal metaphysis were also significantly greater in the xylitol-fed group than in the non-xylitol group. The total bone mineral density and the total bone mineral content of the femoral neck in the xylitol-fed aged group significantly exceeded those in the aged group without xylitol supplementation. CONCLUSIONS: A continuous moderate dietary xylitol supplementation leads to increased bone volume and increased bone mineral content in the long bones of aged rats. This indicates a xylitol-induced protection against aging-related osteoporotic changes.     

Novel bromomelatonin derivatives as potentially effective drugs to treat bone diseases

Several reports indicate that melatonin is involved in the regulation of bone metabolism. To examine the direct effect of melatonin on osteoclasts and osteoblasts, we developed an in vitro assay using fish scales that contain osteoclasts, osteoblasts, and bone matrix, all of which are similar to those found in mammalian membrane bone. Using the assay, we demonstrated that melatonin suppressed osteoclastic and osteoblastic activities. These findings are in agreement with the reports from in vivo studies in mice and rats. In an attempt to develop molecules that increase bone mass, novel bromomelatonin derivatives were synthesized, and the effects of these chemicals on osteoclasts and osteoblasts using the scale assay were examined. As a result, novel bromomelatonin derivatives with the ability to possibly increase bone formation were identified. In scale osteoclasts, particularly, 1-benzyl-2,4,6-tribromomelatonin had a more potent activity than melatonin. In reference to osteoblasts, this agent (10(-9)-10(-6)M) significantly activated osteoblasts. The effect of 1-benzyl-2,4,6-tribromomelatonin on bone formation was confirmed in ovariectomized rats. Thus, the oral administration of 1-benzyl-2,4,6-tribromomelatonin augmented the total bone mineral density of the femoral metaphysis of ovariectomized rats. The stress-strain index of the diaphysis in 1-benzyl-2,4,6-tribromomelatonin-treated rats significantly increased in comparison with that in ovariectomized rats. In rats fed a low-calcium diet, the total bone mineral density of the femoral metaphysis significantly increased following the oral administration of 1-benzyl-2,4,6-tribromomelatonin. These studies identified a melatonin derivative that may have potential use in the treatment of bone diseases, such as osteoporosis.     

柚皮苷联合运动对去势大鼠骨量和骨强度的影响

目的观察柚皮苷（NG）联合中强度跑台运动对去势大鼠骨质疏松模型的治疗效果。方法2月龄雌性SD大鼠80只,用随机数字法分为假手术组（SHAM）、去势组（OVX）、去势＋不同浓度（40、100、200 mg/kg）柚皮苷组（OVX＋NG）、去势＋跑台＋柚皮苷组（OVX＋EX＋NG）、去势＋跑台组（OVX＋EX）、雌激素组（OVX＋E2）,每组10只。大鼠切除卵巢2个月后开始给药,给药60 d后观察各组大鼠股骨骨密度（BMD）,Micro-CT参数,血清生化指标及及股骨力学性能与组织学改变。结果柚皮苷与跑台运动干预60 d后,OVX＋EX＋NG组大鼠股骨颈力学强度、股骨BMD、BV/TV、Tb.N、Tb.Th等均高于单纯去势组（P〈0.05）,OVX＋EX＋NG组治疗效果与柚皮苷浓度呈正相关,以200 mg/kg柚皮苷效果最佳;200 mg/kg柚皮苷联合中强度跑台运动可进一步提高治疗效果。OVX＋NG组大鼠血清骨钙素升高,Ⅰ型胶原C端肽（CTX-1）降低;OVX＋EX组大鼠骨钙素与CTX-1均降低;OVX＋EX＋NG组大鼠骨钙素水平与柚皮苷组（200 mg/kg）无差异,但高于OVX＋EX。OVX＋EX＋NG组CTX-1水平低于单纯柚皮苷组和单纯跑台组（P〈0.05）。结论柚皮苷联合中强度跑台运动可提高去势大鼠股骨BMD,增加骨小梁数目,改善骨代谢指标,提高股骨力学性能。     

Bone effects of hexarelin,a GH-releasing peptide,in female rats: influence of estrogen milieu

OBJECTIVE: The present study was performed to evaluate the potential influence of the estrogen milieu in modulating the effects of GH/IGF stimulation by a GH-releasing peptide, hexarelin (HEXA), on bone metabolism and mineral density in middle-aged female rats. METHODS: HEXA was administered for 60 days (50 microg/kg s.c. twice a day) to intact and ovariectomized (OVX) 11-month-old female rats and changes in bone parameters were evaluated with respect to those of the same rats under baseline conditions and with those of control rats (intact and OVX) administered isovolumetric amounts of physiological saline. Serum total alkaline phosphatase (ALP) and urinary deoxypyridinoline (Dpd) were measured before and at various times during HEXA treatment. Bone mineral content (BMC) and density of lumbar vertebrae and femoral mid-diaphyses were measured by dual energy X-ray absorptiometry before and after treatment. In all groups, serum IGF-I levels were determined before and during treatment and the GH secretory response to HEXA was assessed at the end of the experiment. RESULTS: In intact rats, HEXA did not modify Dpd urinary excretion, induced a trend toward an increase of serum ALP activity and significantly increased BMC (+6.5%) and bone area (+4.1%) only at lumbar vertebrae. In OVX rats, HEXA did not modify the OVX-induced increase in bone turnover markers (Dpd and ALP) and did not affect the OVX-induced vertebral bone loss, but significantly increased BMC (+7.2%) and bone area (+5.3%) at femoral mid-diaphyses. HEXA significantly increased serum IGF-I levels at day 14, but not at day 60, in both intact and OVX rats, whereas the GH secretory response to HEXA was higher in the former than in the latter. CONCLUSIONS: Overall, the present data demonstrate that chronic HEXA treatment increases BMC and bone area at lumbar vertebrae in intact rats and at femoral diaphyses in OVX rats. The different sensitivity to HEXA of the skeletal districts examined is related to the estrogen milieu and may reflect a complex interplay between estrogens and GH/IGF function.