补肾活血颗粒对骨质疏松大鼠骨组织中OPG/RANKLmRNA及蛋白表达的影响

目的探讨补肾活血颗粒对骨质疏松(OP)大鼠骨组织中骨保护蛋白/核因子-β受体活化因子配体(OPG/RANKL)mRNA及蛋白表达的影响。方法采用卵巢摘除法建立SD大鼠OP模型。分别给予生理盐水(模型组)、50 mg/kg补肾活血颗粒(低剂量组)、100 mg/kg补肾活血颗粒(中剂量组)、200 mg/kg补肾活血颗粒(高剂量组)、30μg/kg 17β-雌二醇(雌激素组)干预治疗12 w,设立假手术组为对照。比较各组骨密度(BMD)、骨组织中OPG/RANKL mRNA及蛋白表达水平。结果与假手术组比较,模型组大鼠BMD、OPG/RANKL mRNA及蛋白表达显著减低(P0.05)。与模型组比较,低、中、高剂量组及雌激素组BMD、OPG/RANKL mRNA及蛋白表达明显提高(P0.05)。高剂量组BMD、OPG/RANKL mRNA及蛋白表达显著高于低、中剂量组及雌激素组(P0.05)。结论补肾活血颗粒可有效改善OP大鼠骨密度,调节OPG/RANKL可能是其作用机制。     

淫羊藿甙对去卵巢大鼠骨组织核心结合因子α1表达的影响

目的 观察淫羊藿甙对去卵巢大鼠骨组织Cbfa1表达的影响.方法 54只雌性SD大鼠随机分成6组9只/组:假手术组、模型组、尼尔雌醇组、淫羊藿甙低剂量组、淫羊藿甙中剂量组和淫羊藿甙高剂量组.对尼尔雌醇组和淫羊藿甙低、中、高剂量组分别给予尼尔雌醇(0.1mg/kg)和淫羊藿甙(40,80和160mg/kg)治疗12周.12周后,以DXA法测定大鼠全身的骨密度;放射免疫法测定各组大鼠血清中骨钙素及雌二醇的浓度;处死各组大鼠,直接从头盖骨中提取总.RNA,采用荧光定量PCR技术检测Cbfa1 mRNA的相对表达量.结果 12周后,模型组大鼠的骨密度和血清雌二醇水平明显降低,与假手术组相比,差异有显著性(P<0.05).与模型组相比高剂量组的全身骨密度和血清骨钙素含量增加(P<0.05),但是血清雌二醇的水平无变化(P>0.05).12周后,与假手术组相比,模型组大鼠骨组织中Cbfal mRNA表达明显降低,差异有显著性(P<0.01).与模型组相比高剂量组骨组织中Cbfal mRNA的表达,显著升高(P<0.01).结论 淫羊藿甙对去卵巢大鼠骨质疏松症具有防治作用,其部分机制可能为淫羊藿甙促进骨组织中Cbfa1表达,从而调节骨形成.     

番茄红素对去卵巢大鼠骨质保护作用的研究

目的 研究去卵巢大鼠体内氧化应激水平和骨质的变化及番茄红素对其影响.方法 将32只雌性SD大鼠随机分成4组:假手术组,模型组,小剂量干预组和大剂量干预组.术后1 w开始用番茄红素灌胃,于术后10 w时处死各组大鼠,并对血生化、氧化及抗氧化指标,骨密度,生物力学等参数进行检测,对其骨组织形态学进行观察.结果 在去势10 w后,与假手术组相比,模型组大鼠血清钙、磷、碱性磷酸酶、丙二醛等均不同程度的升高(P＜0.01,P＜0.05),而血清超氧化物歧化酶水平、骨密度值及生物力学指标均不同程度的下降(P＜0.01,P＜0.05).药物干预组,尤其是大剂量干预组的上述参数指标与模型组相比呈现出显著差异(P＜0.01,P＜0.05).结论 氧化应激可能与骨质疏松的发生相关,并初步证实番茄红素对去势大鼠骨质具有保护作用.     

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。     

全身振动对去卵巢大鼠骨组织血管内皮生长因子蛋白表达的影响

目的观察全身垂直振动对去卵巢骨质疏松大鼠骨密度和骨组织血管内皮生长因子(VEGF)蛋白表达的影响。方法将36只3月龄未经产雌性SD大鼠,按体重分层后随机分为假手术组(Sham,12只)、去卵巢静止组(OVX,12只)和去卵巢振动组(WBV,12只)。去卵巢手术10 w时,去卵巢振动组大鼠开始在振动台上接受振动频率为90 Hz振动加速度为3.8 g的全身垂直振动刺激,振动2次/d,15 min/次,振动7 d/w。振动处理8 w时麻醉大鼠,用XR-36型双能X线骨密度仪(DEXA)检测在体骨密度后,腹主动脉取血处死各组大鼠,游离第3腰椎以及左后肢股骨和胫骨,Western印迹法检测第3腰椎、股骨远端和胫骨近端骨组织VEGF蛋白的表达量。结果①在体骨密度检测结果表明,去卵巢静止组第5腰椎、股骨远端和胫骨近端骨密度显著低于假手术组(P<0.01或P<0.001),而全身骨密度和股骨近端骨密度无显著性差异(P>0.05);去卵巢振动组胫骨近端骨密度显著高于去卵巢静止组(P<0.01),而其他部位骨密度无显著性差异(P<0.05)。②与假手术组比较,去卵巢静止组胫骨近端VEGF蛋白表达显著降低(P<0.05),而股骨远端和第3腰椎差异无显著性(P>0.05);与去卵巢静止组比较,去卵巢振动组第3腰椎、股骨远端和胫骨近端VEGF蛋白表达均显著增加(P<0.05或P<0.01)。结论全身垂直振动能显著增加去卵巢骨质疏松大鼠骨组织VEGF蛋白的表达。     

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.     

温阳益气、活血利水法对冠脉结扎心肌梗死后心衰大鼠NT-proBNP的影响

目的 观察温阳益气、活血利水法对心肌梗死后心衰大鼠N-末端脑钠肽前体(NT-proBNP)的影响.方法 结扎大鼠冠状动脉左前降支,制作大鼠心肌梗死后心衰模型,分为对照组,假手术组,模型组,温阳益气、活血利水法组方高、中、低剂量组,卡托普利组.中药干预后,检测大鼠血清NT-proBNP水平.结果 各模型组血清NT-proBNP水平均显著高于假手术组(P＜0.01);与模型组比较,温阳益气、活血利水法组方各剂量组血清NT-proBNP水平均降低(P＜0.05或P＜0.01);卡托普利组血清NT-proBNP水平与温阳益气、活血利水法组方高剂量组相近(P＞0.05);空白对照组和假手术组比较,血清NT-proBNP水平无显著性差异.结论 温阳益气、活血利水法可降低心肌梗死后心衰大鼠血清NT-proBNP水平.     

健心颗粒对早期心力衰竭大鼠心肌组织TNF-α、IL-6的影响

目的探讨健心颗粒治疗早期充血性心力衰竭(CHF)的作用机制。方法采用腹主动脉缩窄术制作压力超负荷慢性早期心力衰竭大鼠模型,实验分组为假手术组、模型组、中药高剂量组、中药低剂量组、西药组。以中药健心颗粒高、低剂量和贝那普利(洛汀新)进行干预,测定各组大鼠心肌组织肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)的含量。结果模型组心肌组织TNF-α、IL-6含量明显高于假手术组(P0.01);西药组、中药低剂量组、中药高剂量组TNF-α、IL-6含量均低于模型组(P0.05或P0.01),中药高剂量组TNF-α、IL-6含量低于西药组、中药低剂量组(P0.05或P0.01)。结论 TNF-α、IL-6在CHF的病理过程中起着重要作用,健心颗粒可通过抑制TNF-α、IL-6的分泌和表达,减轻心肌损伤程度,阻断心室重构过程,从而改善心功能,发挥抗CHF的作用。     

地黄对去卵巢骨质疏松大鼠腰椎骨组织整合素β1 mRNA表达的影响

目的探讨地黄对去卵巢骨质疏松大鼠股骨骨密度、腰椎骨组织整合素β1 mRNA表达的影响。方法 72只雌性大鼠,随机分为假手术组、模型组、雌二醇组和地黄小、中、大剂量组。假手术组仅行假手术,其余五组行卵巢切除术。术后1w分别灌胃给予17β-雌二醇和地黄小、中、大剂量,连续给药3个月。测定血钙、磷和碱性磷酸酶(ALP),尿钙(u-Ca)、磷、尿脱氧吡啶酚(D-Pyr)和肌酐(Cr)。之后处死动物,取出右侧股骨,测定骨密度;取出第2腰椎,测定骨钙(b-Ca)、磷(b-P)含量;取出第4腰椎,采用实时荧光逆转录聚合酶链反应测定腰椎骨整合素β1 mRNA表达。结果与假手术组相比,模型组血清ALP、u-Ca、D-Pyr/Cr显著增加,股骨密度、腰椎骨整合素β1 mRNA表达均降低。与模型组比较,地黄中、大剂量组和雌二醇组均可使血清ALP、u-Ca、D-Pyr/Cr排出量降低,股骨密度、腰椎骨整合素β1 m RNA表达均增加。结论地黄能抑制由于去卵巢雌激素缺乏引发的骨转换增强,提高骨密度,促进腰椎骨组织整合素β1 m RNA表达,增强骨质量。     

Sex differences in cancellous and cortical bone strength, bone mineral content and bone density.

The purpose of this study was to examine sex differences in cancellous and cortical bone strength, bone mineral content (BMC) and bone density of excised cadaver vertebral and phalangeal bones. The samples were age-matched. Bone strength was measured as the mechanical force required to crush or break the bones. Two parameters of bone strength were used on the vertebrae; the force at the first deviation from linearity and the mean force during the consolidation before final failure. The force at first deviation from linearity was not significantly different between the sexes, but there was a significant difference in the consolidation force. The mean men's phalangeal strength was twice that of the women's. BMC and BMC/BW of both types of bone were statistically different between the sexes. Radiographic photodensity measures on the vertebrae showed no sex differences. Cortical diameters of the finger bones were significantly greater in males.     

Mechanisms of bone resorption and new bone formation in spondyloarthropathies

Spondyloarthropathies (SpA) share clinical features such as sacroiliitis, axial immobility, and peripheral arthropathies. They also share a strong association with human leukocyte antigen-B27, implicating T cells and antigen-presenting cells in the disease process. Inflammation seems to underlie the pathogenesis of SpA, particularly in the axial skeleton and entheses. Pathologic bone loss and formation occur simultaneously in inflamed regions, suggesting an inflammation-induced dysregulation of osteoclast and osteoblast activity. Pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNFα) appear to be central to the disease, because TNFα blockade has been shown to effectively improve clinical outcome. Other cytokines such as transforming growth factor-beta, interferon-gamma (IFNγ), and interleukin-18 are also likely to be important in SpA. Activated T cells have been shown to produce cytokines such as IFNγ and receptor activator of nuclear-factorkappaB ligand, with direct effects on osteoclastogenesis. The dual role of T cells in immunobiology and skeletal biology provides a possible link between human leukocyte antigen-B27, pro-inflammatory cytokines, and bone cells in SpA.     

Metabolic bone disease and bone quality

On the progress of study concerned with pathology of metabolic bone disease such as osteoporosis, it has been known that most of bone strength can be explained by bone volume. As bone volume can be determine by bone mineral density (BMD) with dual-energy X-ray absorptiometry (DXA), it has been widely used for diagnosis of osteoporosis or efficacy of treatment. However, with the advance of bone morphometry, decrease of bone strength or existence of insufficiency fracture is influenced by not only loss of BMD but also deterioration of bone quality especially bone microstructure. In this chapter, we will give an outline of change of bone quality in metabolic bone disease.     

The role of cortical bone and its microstructure in bone strength

Bone's mechanical competence and its fragility in particular depend to a certain extent on the structure and microstructure of the cortical bone compartment. Beyond bone mineral density (BMD) and bone mineral content, a variety of other features of cortical bone contribute to whole bone's resistance to fracture. Structural properties of cortical bone most commonly employed as surrogate for its mechanical competence include thickness of the cortex, cortical cross-sectional area and area moment of inertia. But microstructural properties such as cortical porosity, crystallinity or the presence of microcracks also contribute to bone's mechanical competence. Microcracks in particular not only weaken the cortical bone tissue but also provide an effective mechanism for energy dissipation. Bone is a damageable, viscoelastic composite and most of all a living material capable of self-repair and thus exhibits a complex repertoire of mechanical properties. This review provides an overview of a variety of features of cortical bone known to provide mechanical competence and how these features may be applied for fracture risk prediction.     

Effect of age on bone density and bone turnover in men

OBJECTIVE Little Is known about the pattern of age-related bone loss in men, and although androgens are required for optimum bone mass it Is not clear whether the fall in bone mass with age in men is related to failing androgens. DESIGN Cross-sectional measurement of bone density, at five sites, and markers of bone resorption and formation in 147 normal volunteers aged 20-83 years. SUBJECTS Healthy laboratory workers, hospital staff, their relatives, and husbands of women attending our osteoporosis clinic. MEASUREMENTS Forearm density (fat corrected), spine L2-L4, femoral neck, Ward's triangle and trochanter density; serum procollagen I C-terminal extension peptide, osteocalcin, bone alkaline phosphatase and collagen I C-terminal telopeptlde; fasting urine hydroxy-proline/creatinine, pyridinoline/creatinine and deoxy-pyridinoline/creatinine; and free androgen Index (FAI), measured as serum testosterone/sex hormone binding globulin. RESULTS Bone loss accelerated at most sites after age 50. There was a significant fall In FAI from the third decade onwards. The levels of ail bone markers fell with age. CONCLUSIONS Bone loss In men appears to accelerate from age 50 and is associated with decreased bone formation which may be associated with falling levels of free androgen.     

Dissociation between bone resorption and bone formation in osteopenic transgenic mice

Bone mass is maintained constant in vertebrates through bone remodeling (BR). BR is characterized by osteoclastic resorption of preexisting bone followed by de novo bone formation by osteoblasts. This sequence of events and the fact that bone mass remains constant in physiological situation lead to the assumption that resorption and formation are regulated by each other during BR. Recent evidence shows that cells of the osteoblastic lineage are involved in osteoclast differentiation. However, the existence of a functional link between the two activities, formation and resorption, has never been shown in vivo. To define the role of bone formation in the control of bone resorption, we generated an inducible osteoblast ablation mouse model. These mice developed a reversible osteopenia. Functional analyses showed that in the absence of bone formation, bone resorption continued to occur normally, leading to an osteoporosis of controllable severity, whose appearance could be prevented by an antiresorptive agent. This study establishes that bone formation and/or bone mass do not control the extent of bone resorption in vivo.     

Vanadate stimulates bone cell proliferation and bone collagen synthesis in vitro

We recently proposed a hypothesis for the molecular mechanism of the osteogenic action of fluoride in which it stimulates osteoblast proliferation via the inhibition of an osteoblastic acid phosphatase-like phosphotyrosyl protein phosphatase activity. To test this hypothesis, we investigated whether orthovanadate, a known phosphotyrosyl protein phosphatase inhibitor, would mimic fluoride in the stimulation of bone cell proliferation and bone collagen synthesis in vitro. Orthovanadate inhibited the osteoblastic acid phosphatase activity and stimulated bone cell proliferation at the same low concentrations (i.e. 5-15 microM). At the mitogenic doses, orthovanadate also showed a dose-dependent increase in alkaline phosphatase (a marker of mature osteoblasts) in cultured calvarial cells and stimulated bone collagen synthesis, as measured by the incorporation of [3H]proline and the conversion into [3H] hydroxyproline in organ calvaria cultures. Therefore, orthovanadate stimulated bone formation by increasing the number of mature osteoblasts mediated via stimulation of cell proliferation and differentiation. Orthovanadate was dependent on the presence of a mitogen in cell medium for its mitogenic action in vitro and synergistically potentiated the mitogenic actions on osteoblasts of those growth factors, i.e. insulin, epidermal growth factor, insulin-like growth factor I, and skeletal growth factor, whose mitogenic action involved tyrosyl protein phosphorylation. However, the interaction between orthovanadate and basic fibroblast growth factor, a growth factor that does not appear to involve tyrosyl protein phosphorylation, on bone cell proliferation was additive. In summary, these data are consistent with the hypothesis that inhibition of the osteoblastic phosphotyrosyl protein phosphatases can prolong and/or potentiate the mitogenic actions of growth factors, and thereby stimulates cell proliferation.     

Deconditioning increases bone resorption and decreases bone formation in the rat

The response of calcium metabolism and bone turnover to deconditioning after exercise training was studied in three groups of Sprague-Dawley female rats: age-matched controls, 12-week treadmill training, and 8-week treadmill training followed by 4-week discontinuation of training. The exercised rats had a higher absorption efficiency of calcium that decreased to control levels as early as 2 weeks following discontinuation of training. Urinary calcium excretion increased by 2 weeks of deconditioning and then reverted to control levels; bone mineral content measurements declined following deconditioning. Tracer uptake of 45Ca in the femur demonstrated an increase in bone formation that was present during exercise, but was not evident following 4 weeks of deconditioning. The urinary excretion of prelabeled 3H-tetracycline was used to investigate bone resorption in two groups of exercised rats; one group continued to exercise and in the other group the exercise was discontinued. Bone resorption increased within 4 to 11 days after deconditioning and returned to basal levels by the 10th day, at which time the net bone retention of 3H-tetracycline was 86% of that of the group that continued exercising. We conclude that exercise must be continued to sustain any gain it produces in bone mineral. Bone mass gained through exercise will be lost during deconditioning as a result of a decline in bone formation and an increase in bone resorption.     

Metabolic bone markers and bone cell biology

Various metabolic bone markers have been developed in order to analyze each process of bone resorption and bone formation. By evaluating the two processes using bone markers, an imbalance between bone resorption and formation can be estimated. Metabolic bone markers have already been in clinical use for the early diagnosis and the assessment of treatment efficacy in osteoporotic patients and for the diagnosis of cancer-induced bone diseases. Further elucidation of the mechanisms of formation and secretion, metabolic clearance, diurnal rhythm as well as their changes in various disorders should enable us to evaluate bone turnover at a real-time scale and to utilize for the diagnosis of a variety of metabolic bone diseases.