破骨细胞相关的分化因子与风湿性疾病的骨代谢

骨是一种动态的组织，骨的代谢是骨吸收和与之相伴随的骨形成过程。这一过程维持着骨的平衡，该平衡的破坏，将导致骨量的减少——骨质疏松(osteoporosis)或骨量的增多——骨质硬化(osteopetrosis)。破骨细胞(osteoclasts，OC)来源于骨髓中的单核巨噬细胞集落形成单位(granulocyte     

破骨细胞的雌激素受体

研究表明 激素除可通过细胞因子介导抑制骨吸收外,还可通过雌激素受体介导直接抑制骨吸收,本文综述了骨中破骨细胞受体以及雌激素对破骨细胞作用的受体介导机制。     

RANKL诱导破骨细胞前体细胞分化成熟

目的 用核因子-κB受体活化因子配体(RANKL)诱导破骨细胞前体细胞分化成熟,建立获取成熟破骨细胞的方法.方法 用破骨细胞前体细胞RAW264.7细胞为模型,RANKL诱导培养4～9 d,抗酒石酸酸性磷酸酶(TRAP)染色观察TRAP阳性多核细胞形成,罗丹明-鬼笔环肽荧光染色观察纤维性肌动蛋白(F-actin)环,DAPI染色观察细胞核,甲苯胺蓝染色观察牛骨片表面的吸收陷窝情况.结果 RANKL可诱导RAW264.7细胞形成TRAP染色阳性的多核细胞,形成F-actin环,骨片吸收陷窝明显.结论 RANKL可诱导RAW264.7细胞向成熟破骨细胞分化,该诱导模型可用于破骨细胞分化研究.     

骨质疏松症与破骨细胞性骨吸收

骨质疏松症是以骨量减少，骨组织微观结构退化为特征，以致骨的脆性增高而骨折危险性增加的一种全身性骨病，骨质疏松症就是由于骨代谢的失衡，破骨量大于成骨量所致，破骨细胞的分化或其功能的改变所导致的骨改建失衡是骨质疏松的重要病理基础，许多细胞因子在破骨细胞生成和骨吸收中起作用，破骨细胞骨吸收功能活跃是其发生的病理生理之一，对破骨细胞骨吸收功能的抑制是防治骨质疏松症的主要药理基础。     

破骨细胞在类风湿关节炎骨破坏中的作用研究进展

类风湿关节炎(RA)是一种慢性自身免疫性疾病,可引起关节疼痛、肿胀、僵硬,永久性关节破坏,严重损害时可导致残疾和畸形,其主要特征是关节滑膜的炎症和软骨、骨进行性破坏[1].骨质破坏及由此产生的功能障碍是RA致残的主要原因,多种细胞参与该过程[2],其中破骨细胞在RA骨破坏中发挥关键性的作用[3],其作用过程主要包括:破骨前体细胞的迁移、分化和破骨细胞的骨吸收.研究发现,RA病程中破骨细胞受到一系列调控因素作用,了解这些调控机制,有助于我们寻找到干预RA骨破坏的作用靶点.现就破骨细胞在RA骨破坏中的作用研究进展综述如下.     

破骨细胞及其功能的局部调控

综述了参与破骨细胞开发和影响其功能状态的局部因子，以及破骨细胞骨吸收过程中的一些分子机制的研究进展。影响破骨细胞功能的局部因子有两类：其中，集刺激因子、白介素１、白介素６、淋巴毒素、肿瘤坏死因子－α转化生长因子－α为刺激骨吸收的细胞因子，而干扰素、转化因子－β和白介素－４则有抑制骨吸收的效应。质子泵，粘附蛋白和Ｃ－Ｓｒｃ酪氨酸激酶在破骨细胞发生和功能中作用的阐明为加深对人谢骨病发生机制的认识和开发     

破骨细胞凋亡与骨代谢

细胞凋亡又称细胞程序性死亡 ,是机体为维持内环境稳定 ,由基因控制的细胞自主的有序性死亡。随着细胞生物学、遗传学、分子生物学的发展 ,发现细胞凋亡与多细胞有机体的个体发育、调控、正常生理活动的维持、某些疾病的发生及其细胞恶变均有密切关系〔1〕。近年研究表明 ,破骨细胞凋亡在骨代谢调控和绝经后骨质疏松症等骨代谢疾病的发生、发展过程中具有重要意义。　　一、破骨细胞凋亡表现　　骨的新陈代谢表现为旧骨不断被吸收 ,新骨随之持续形成 ,即骨重建。承担骨重建的基本多细胞单位 (ｂａｓｉｃｍｕｌｔｉｃｅｌｌｕｌａｒｕｎｉｔ,…     

降钙素对破骨细胞骨吸收抑制作用的机理

降钙素对破骨细胞骨吸收抑制作用的机理于明香王洪复破骨细胞（ｏｓｔｅｏｃｌａｓｔ，ＯＣ）是一种多核的骨吸收细胞，在骨再建过程中起着启动和先锋作用，与成骨细胞通过相互调控机制，共同完成骨的吸收与形成，使骨组织不断更新。破骨细胞功能过度活跃时，骨吸收亢进，...     

破骨细胞生成抑制因子及分化因子的研究进展

越来越多的研究资料表明破骨细胞分化因子／护骨因子配基可能是唯一能够直接诱导破骨细胞分化的细胞分子,其功能性受体RANK位于前破骨细胞及破骨细胞表面,介导了细胞分化信号的传导,破骨细胞生成抑制因子／护骨因子是破骨细胞分化因子／护骨因子配基的假活性受体,能竞争性抑制破骨细胞分化因子／护骨因子配基与RANK的结合,从而抑制破骨细胞的生成及功能。本文综述了破骨细胞生成抑制因子和分化因子的发现,结构和功能,并探讨了二者之间的相互作用机制。     

源于脾干细胞的破骨细胞诱导生成及培养

目的建立一种可以获得大量破骨细胞或破骨样细胞的方法，以便进行原发性骨质疏松有关破骨细胞的研究。方法C57雌性小鼠经尾静脉注射5-氟脲嘧啶后，取脾细胞悬液在含有rmIL-3，rhIL-6和胎牛血清(FCS)以及牛血清白蛋白(BSA)的α-MEM-琼脂半固体培养7d左右，获得脾脏造血干细胞集落；此集落再以粒细胞-巨噬细胞集落刺激因子(GM—CSF)诱导生成破骨样细胞。结果经活体观察、酒石酸-抗酸性磷酸酶(TRAP)染色、扫描电镜检查，证实所得细胞为破骨细胞。结论建立了一种获得大量破骨细胞或破骨样细胞方法，即源于脾干细胞的破骨样细胞的诱导生成及其培养。     

Predictors of osteoclast activity in patients with sickle cell disease

Background

Bone changes are common in sickle cell disease, but the pathogenesis is not fully understood. Tartrate-resistant acid phosphatase (TRACP) type 5b is produced by bone-resorbing osteoclasts. In other forms of hemolytic anemia, increased iron stores are associated with osteoporosis. We hypothesized that transfusional iron overload would be associated with increased osteoclast activity in patients with sickle cell disease.

Design and Methods

We examined tartrate-resistant acid phosphatase 5b concentrations in patients with sickle cell disease and normal controls of similar age and sex distribution at steady state. Serum tartrate-resistant acid phosphatase 5b concentration was measured using an immunocapture enzyme assay and plasma concentrations of other cytokines were assayed using the Bio-Plex suspension array system. Tricuspid regurgitation velocity, an indirect measure of systolic pulmonary artery pressure, was determined by echocardiography.

Results

Tartrate-resistant acid phosphatase 5b concentrations were higher in 58 adults with sickle cell disease than in 22 controls (medians of 4.4 versus 2.4 U/L, respectively; P=0.0001). Among the patients with sickle cell disease, tartrate-resistant acid phosphatase 5b independently correlated with blood urea nitrogen (standardized beta=0.40, P=0.003), interleukin-8 (standardized beta=0.30, P=0.020), and chemokine C-C motif ligand 5 (standardized beta=−0.28, P=0.031) concentrations, but not with serum ferritin concentration. Frequent blood transfusions (>10 units in life time) were not associated with higher tartrate-resistant acid phosphatase 5b levels in multivariate analysis. There were strong correlations among tartrate-resistant acid phosphatase 5b, alkaline phosphatase and tricuspid regurgitation velocity (r>0.35, P<0.001).

Conclusions

Patients with sickle cell disease have increased osteoclast activity as reflected by serum tartrate-resistant acid phosphatase 5b concentrations. Our results may support a potential role of inflammation rather than increased iron stores in stimulating osteoclast activity in sickle cell disease. The positive relationships among tartrate-resistant acid phosphatase 5b, alkaline phosphatase and tricuspid regurgitation velocity raise the possibility of a common pathway in the pulmonary and bone complications of sickle cell disease.     

Generation of human osteoclasts in stromal cell-free and stromal cell-rich cultures: differences in osteoclast CD11c/CD18 integrin expression

Osteoclasts form in the presence of macrophage colony-stimulating factor (M-CSF) and receptor activator of Nfkappab ligand (RANKL), a membrane-bound differentiation factor that is now available as a soluble recombinant molecule. Acquisition of the osteoclast phenotype [the alphavbeta3 subunit of the vitronectin receptor (VNR)-, calcitonin receptor (CTR)- and F-actin ring-positive cells] is associated with loss of monocyte/macrophage-associated integrins, specifically CD11b, CD11c and CD18. We hypothesized that differences in the osteoclast integrin adhesion molecule profile may exist in osteoclasts generated in stromal cell-rich and in stromal-free conditions. Unlike osteoclasts generated in vivo, F-actin ring-positive (resorbing) osteoclasts formed in soluble RANKL in vitro, in the absence of stromal cells, and co-expressed CD11c and CD18. However, when osteoclasts were generated from peripheral blood mononuclear cells (PBMNCs) in co-cultures with the murine bone marrow stromal cell line 218 (which does not produce membrane-bound RANKL) in the presence of soluble RANKL, CD11c and CD18 were not expressed by osteoclasts. These findings indicate that the persistent expression of CD11c and CD18 is not accounted for by RANKL being presented in a soluble form and that membrane-bound RANKL is not required for the normal integrin expression in resorbing osteoclasts. This study demonstrates that potentially misleading information may arise by using data obtained from osteoclasts generated in the absence of stromal cells as they do not completely reflect the situation in vivo.     

右归饮对类固醇激素性股骨头坏死大鼠成骨-破骨体外共育体系中破骨细胞分化的影响

目的 探讨右归饮对激素性股骨头坏死（SINFH）大鼠成骨-破骨体外共育体系中破骨细胞（osteoclast,OC）生长分化的影响.方法 雄性SD大鼠70只,体重（100±20）g,用随机数字表法取30只用醋酸泼尼松龙49 mg/kg·d肌内注射造模,1周后随机抽取2只造模大鼠进行组织病理观察确定造模成功.40只正常大鼠随机分为高、中、低右归饮给药组及蒸馏水组.从造模大鼠股骨、胫骨分离培养成骨细胞（osteoblast,OB）和OC诱导5 d,将OB以1×105/mL密度接种于成骨-破骨细胞共育体系中,分别以蒸馏水血清（对照组）、低（0.5 mL/100 g）、中（1.0 mL/100 g）、高（2.0 mL/100 g）浓度右归饮含药血清作用于OB-OC共育体系.用抗酒石酸酸性磷酸酶（TRAP）鉴定OC和阳性多核细胞计数,用RT-PCR方法测定RANKL mRNA表达水平变化.结果 与蒸馏水组比较,高、中浓度右归饮组TRAP染色阳性破骨细胞数量均显著减少（P＜0.01）;RANKL基因表达水平明显降低（P＜0.01）,高浓度右归饮组与低、中浓度组比较RANKL基因表达水平显著降低（P＜0.01、P＜0.05）.结论 一定浓度的右归饮对SINFH大鼠体外成骨-破骨共育体系破骨细胞形成、分化有抑制作用,以高浓度右归饮作用最强.     

氟对大鼠破骨细胞基质金属蛋白酶-9的影响

目的 观察氟对大鼠破骨细胞分泌的基质金属蛋白酶-9(MMP-9)的影响。方法 采用western blot技术观察染氟大鼠股骨干骺端MMP-9的表达；应用免疫组织化学方法观察氟对体外培养的破骨细胞MMP-9的表达的影响。结果 染氟大鼠股骨干骺端MMP-9的蛋白表达增高；氟可使体外培养的破骨细胞MMP-9的蛋白表达增高。结论 氟通过提高破骨细胞的MMP-9的表达而致破骨性骨吸收活性增强。     

铁超载对小鼠单核细胞RAW264．7向破骨细胞分化的影响

]目的探讨铁超载对小鼠单核细胞RAW264．7向成熟破骨细胞分化的影响。方法实验分为正常对照组、低铁对照组、高铁干预组3组,用含有核因子KB受体活化因子配体（RANKL）和巨噬细胞集落刺激因子（M—CSF）的培养基诱导RAW264．7向破骨细胞分化,在此过程中加入去铁胺（DFO）和不同浓度的枸橼酸铁铵（FAC）。对培养第4天和第7天的细胞进行抗酒石酸酸性磷酸酶（TRAP）染色,对TRAP阳性细胞进行计数,用酶联免疫吸附法（ELISA）检测培养基中TRAP-Sb的含量。结果（1）高铁干预组的TRAP阳性细胞数高于正常对照组和低铁对照组（P〈0．05）,具有时间和浓度依赖性;（2）高铁干预组的TRAP-Sb的含量高于正常对照组（P〈0．05）,具有浓度依赖性。结论铁超载能促进RAW264．7向破骨细胞分化。     

人破骨细胞分化因子基因的克隆及在COS-7细胞中的表达

目的 克隆人破骨细胞分化因子(ODF)基因并在真核细胞中表达。方法 以人骨肉瘤细胞系MG63的总RNA为模板，采用反转录-聚合酶链反应(RT-PCR)法得到ODF的编码区cDNA，构建真核表达载体pcDNA3．1( )-ODF，在脂质体介导下转染非洲绿猴肾细胞系(COS)-7，经G418压力筛选建立稳定转染人ODF的细胞系，Western印迹检测其在COS7细胞中的表达。结果获得的人ODFcDNA序列与文献报道的核苷酸序列一致，Western印迹证实稳定转染人ODF的COS7细胞系中有ODF的表达。结论 构建了人破骨细胞分化因子(ODF)的真核表达载体，并在COS7细胞中获得稳定表达。     

Replacement Process of Carbonate Apatite by Alveolar Bone in a Rat Extraction Socket

The objective of this study was to investigate a bone graft substitute containing carbonate apatite (CO₃Ap) to analyze bone replacement and the state of bone formation in vitro and in vivo compared with autogenous bone (AB) or control. An osteoclast precursor cell line was cultured with AB or CO₃Ap, and morphological analysis using scanning electron microscopy and a tartrate-resistant acid phosphatase activity assay were performed. The right maxillary first and second molars of Wistar rats were extracted and compensated by AB or CO₃Ap granules. Following implantation, the bone formation state was evaluated after 3, 5, 7, 14, and 28 days of surgery by micro-computed tomography and immunohistostaining. The osteoclast-like cell morphology was typical with many cell protrusions in the AB and CO₃Ap groups. Additionally, the number of osteoclast-like cells formed in the culture increased in each group; however, there was no significant difference between the AB and CO₃Ap groups. Five days after tooth extraction, osteoclasts were observed near CO₃Ap. The bone thickness in the CO₃Ap group was significantly increased than that in the control group and the bone formation in the CO₃Ap group increased by the same level as that in the AB group. CO₃Ap is gradually absorbed by osteoclasts in the extraction socket and is easily replaced by alveolar bone. The process of bone replacement by osteoclasts is similar to that of autologous bone. By observing the process of bone replacement in more detail, it may be possible to gain a better understanding of the bone formation and control the amount of bone after surgery.