骨保护蛋白配体与骨吸收

骨终生都在进行改建 (Ｒｅｍｏｄｅｌｉｎｇ) ,通过持续不断的骨形成和骨吸收来维持其形态与功能。这一生理过程涉及到成骨细胞 (ＯＢ)和破骨细胞 (ＯＣ)。ＯＣ来自于造血的祖细胞(ｐｒｏｇｅｎｉｔｏｒ)或造血性单核 /巨噬细胞前体 (ｐｒｅｃｕｒｓｏｒ)。这种前体细胞在骨髓微环境和成骨细胞系分泌的细胞因子的作用下 ,在骨吸收的位点分化为成熟的ＯＣ[1～ 3] 。然而从ＯＣ前体分化为成熟的ＯＣ的确切机制目前还不清楚 ,直至最近骨保护蛋白配件 (Ｏｓｔｅｏｐｒｏｔｅｇｅｒｉｎｌｉｇａｎｄ ,ＯＰＧＬ)的发现 ,才使人们对这一问题的认识…     

阿胶强骨口服液含药血清对胎鼠成骨细胞护骨素及护骨素配体mRNA表达的影响

目的研究阿胶强骨口服液含药血清对胎鼠成骨细胞中OPG、RANKL表达的影响,探讨阿胶强骨口服液治疗骨质疏松的分子机制。方法 3月龄Wistar大鼠(雌雄各半)30只,随机分为阿胶强骨口服液,雌激素,及生理盐水对照组,灌胃7d后,制备实验组和对照组的含药血清。将新生SD大鼠分离的颅骨成骨细胞,制成单细胞悬液,消化传代,取二代培养的成骨细胞,制成细胞悬液。培养细胞被分为5组,分别加同体积药液,对照组仅加培养液,继续培养。采用MTT比色分析、3H-胸腺嘧啶核苷(3H-TdR)渗入法测定成骨细胞增殖,用放免法测定细胞内BGP含量,RT-PCR测定胎鼠成骨细胞中OPG、RANKL mRNA的表达。结果阿胶强骨口服液含药血清以剂量依赖方式促进成骨细胞的增殖,与对照组相比,差异显著(P<0．05)。成骨细胞OPG基因mRNA表达在阿胶强骨口服液含药血清100ml／L时最强,与雌激素组比较无显著性差异(P>0 05),且明显高于对照组(P<0．05)。成骨细胞RANKL基因mRNA表达在1000ml／L阿胶强骨口服液含药血清与雌激素组比较无显著性差异(P<0 05),且明显低于对照组(P<0．05)。结论阿胶强骨口服液可在细胞水平上促进骨的合成代谢及前成骨细胞的有丝分裂,分子水平上调节 OPG／RANKL表达而治疗骨质疏松。     

雷洛昔芬上调卵巢切除大鼠腰椎护骨素mRNA的表达

目的建立绝经后骨质疏松（postmenopausal osteoporosis，PMOP）的大鼠模型，研究雷洛昔芬（raloxifene，RLX）对卵巢切除大鼠腰椎组织中护骨素（OPG）基因mRNA表达水平的影响，探讨其治疗PMOP的作用机制。方法6月龄未交配健康雌性SD大鼠40只，随机分为假手术（SHAM）组，卵巢切除（OVX）组，OVX＋戊酸雌二醇（E2）组和OVX＋RLX组。灌胃13周后处死动物，第四腰椎行骨组织形态计量指标测定，第二腰椎采用RT-PCR方法，对OPGmRNA表达水平进行检测。结果OVX组大鼠较SHAM组骨小梁体积百分比（TBV％）显著下降；类骨质体积百分比（OSV％）明显升高（P〈0．05）；B和RLX组大鼠的TBV％明显升高，OSV％明显下降；OPGmRNA表达水平在OVX大鼠组织中下调，与SHAM组相比有显著性差异（P〈0．05），E2和RLX均可上调OVX大鼠骨组织OPGmRNA表达水平（P〈0．05）。结论实验成功制造了大鼠PMOP模型；E2和RLX均可抑制骨转换；PMOP的发生与OPG有关，RLX和E2一样，其抗骨吸收效应很可能是通过OPG介导的。     

骨保护素及其配体

骨重建是破骨细胞 (Osteoclast,OC)骨吸收和成骨细胞 (Osteoblast,OB)骨形成不断进行的过程 ,正常情况下骨吸收和骨形成保持着动态平衡 ,一旦OC数目和活性出现异常 ,就可导致代谢性骨病 (骨硬化和骨质疏松 )。既往研究认为造血前体细胞分化为成熟OC受骨营养激素、微环境中局部生长因子和成骨间充质细胞的影响。近年研究表明 :①OC发育异常、基因突变或激活缺失均可导致骨吸收的增加和严重的骨硬化〔1〕;②间充质微环境对OC发育起重要作用〔2〕;③通过Src酪氨酸激酶的跨膜信号传导是OC介导骨吸收的途径〔3〕;④核因子fos调节基因、M …     

蛇床子素对大鼠成骨细胞中0PG和RANKL基因mRNA表达的影响

目的 观察蛇床子素(OST)对体外培养的大鼠成骨细胞中OPG中RANKL基因表达的影响。方法 新生SD大鼠颅骨成骨细胞培养,设空白对照组和用不同浓度蛇床子素培养液(10-7,10-6,10-5mol／L)培养处理的大鼠成骨细胞为实验组,未经处理的细胞为空白对照组,分别提取48 h和7 d的细胞总mRNA,用RT-PCR技术分析OPG／RANKL的mRNA表达情况进行半定量比较。结果48h两基因都已有表达,相对空白对照组,OST上调OPG mRNA的表达(P<0.05),但对RANKL的表达无显著影响,OPG／RANKL比值变大;7 d时OPG和RANKL表达都有增加,OST上调OPG的表达(P<0.01),OST在10-5mol／L下调RANKL的表达(P<0.05),OPG／RANKL比值增大。结论OST可以增加大鼠成骨细胞OPG的表达,同时轻微抑制RANKL的表达,早期对RANKL的表达影响不大。     

Genistein对破骨细胞分化和骨吸收功能的影响及其机制探讨

目的 探讨三羟异黄酮类药Genistein对破骨细胞性骨吸收的作用及其机制。方法 1，25(OH)2D3诱导小鼠骨髓细胞形成破骨样细胞，分别加入0moL／L、10^-9moL／L、10～moL／L、10^-7moL／L、10～moL／L、10^-5moL／LGenistein，于培养3，5和8d分别计数抗酒石酸酸性磷酸酶阳性细胞个数；于培养12d利用图像分析系统对骨吸收陷窝的面积进行测量分析。另从小鼠颅盖骨分离培养获得原代成骨细胞，分别加入0，10^-8，10^-7，10^-6和10^-5moL／LGenistein并以10^-9moL／L^-7 β雌二醇为对照，采用RT-PCR的方法检测Genistein对骨保护因子(osteoprotegerin，OPG)及破骨细胞分化因子(receptor activator of NF-κB ligand，RANKL)mRNA表达的影响。结果 利用1，25(OH)2D3成功诱导出破骨样细胞；随Genistein浓度增加，抗酒石酸酸性磷酸酶阳性细胞(单核、双核和多核)个数和骨吸收陷窝面积呈剂量依赖性减少。同时，应用Genistein后原代成骨细胞中OPG和RANKL的mRAN表达都有增强，但其最终效应表现为剂量依赖性和时间依赖性地增大OPG／RANKL的浓度比。结论 Genistein通过抑制破骨前体细胞分化来抑制其体外骨吸收功能，其分子机制与OPG／RANKL mRNA表达比值的升高有关。     

护骨素系统与骨质疏松症

骨质疏松症是发病率高、死亡率高、保健费用消耗大的骨代谢疾病。引起骨质疏松的原因多种多样 ,主要有 :( 1)后天获得骨量不足 ;( 2 )遗传基因和基因的多态性。二者最终表现为破骨细胞功能的亢进 ,骨吸收速率超过骨形成而致骨质疏松。新近研究发现破骨细胞虽受多种激素和细胞因子调节 ,而最终效应因子是护骨素 (osteoprotegerin ,OPG)系统。该系统从分子水平阐述了各种骨吸收刺激因子激活破骨细胞的机理 ,开辟了对骨质疏松症防治研究的新方法。1　OPG系统的发现及结构特点1.1　OPG系统的发现1997年Simonet等发现一个基因 ,其编码的蛋白…     

不同浓度钛微粒对护骨素/护骨素配体基因表达影响的体外研究

目的: 探讨磨损微粒引起假体周围骨溶解的机理, 为人工关节假体松动的预防研究打下基础。方法: 采用不同浓度钛微粒 (粒径 <3μm) 对成骨细胞MG 63细胞进行 24h干预, 半定量RT PCR观察其对护骨素 (OPG) 及护骨素配体 (OPG- L) 基因表达的影响。结果:钛微粒能上调MG 63细胞OPG及OPG- L基因表达, 且对后者作用强于前者。结论: OPG- L/OPG比率随着钛微粒浓度的加大而增高, 引起骨溶解大于骨形成, 这可能是磨损微粒引起人工关节假体松动的重要原因, 这表明降低OPG- L/OPG比率有可能预防或逆转此病理过程。     

去势大鼠骨髓体外培养破骨细胞观察及OPG mRNA表达的变化

目的 观察去势大鼠骨髓源性破骨细胞形成的动态变化和骨髓细胞护骨素(OPG)基因的变化.方法 健康Wistar雌性大白鼠60只,分为实验组(去卵巢组)和对照组(假手术组),每组30只,分别于术后2、4、6、8和12周取去卵巢组和对照组每只大鼠股骨骨髓细胞进行细胞培养,左侧股骨培养细胞作瑞氏-吉姆萨染色和抗酒石酸酸性磷酸酶(TRAP)染色,观察破骨细胞并计数.右侧股骨培养细胞提取总RNA,通过RT-RCR来观察去势大鼠骨髓细胞OPG mRNA表达强度变化.结果 术后2、4周去势组破骨细胞形成数多于同期对照组,6周去势组破骨细胞形成达到高峰,显著多于同期对照组P<0.01.8周去势组破骨细胞数较6周有所下降,但仍明显高于对照组.骨髓培养细胞OPG mRNA表达,术后2、4周OPG mRNA表达较同期对照组无明显差别,6周去势组OPG mRNA表达明显低于对照组,8周去势组OPG mRNA表达与同期对照组比较无明显差异.结论 大鼠去势后可引起破骨细胞形成数量增多和OPG mRNA表达降低,且均在术后6周表现明显.     

骨保护素和骨保护素配体在人骨髓基质细胞向成骨细胞分化过程中的表达

目的研究骨保护素和骨保护素配体在人骨髓基质细胞向成骨细胞诱导分化过程中的表达情况，探讨其在骨重建过程中的调节作用。方法实验中采用梯度离心法和酶消化法分别获得人骨髓基质细胞和成骨细胞，并将骨髓基质细胞向成骨细胞方向诱导分化。通过形态学观察、生化指标检测、细胞染色和矿化结节测定等方法，确定骨髓基质细胞的功能状态和分化程度。采用RT-PCR和Westem blot方法，检测骨髓基质细胞向成骨细胞分化过程中骨保护素和骨保护素配体的表达情况。结果获得的骨髓基质细胞和成骨细胞生长状态良好，生化指标稳定。骨髓基质细胞分化后，碱性磷酸酶分泌明显增加，可以产生大量的矿化结节，具有成熟成骨细胞的表型特征。RT-PCR和Western blot检测，在骨髓基质细胞向成骨细胞分化过程中，骨保护素在mRNA和蛋白质水平的表达明显升高，而骨保护素配体的表达则逐渐下降。细胞中OPG mRNA表达在第21天时达到最大，约为未分化时水平的2．5倍。而OPGLmRNA表达减少为未分化时1／2；细胞中OPG的蛋白质表达水平提高约为未分化细胞的6倍。统计学分析，P〈0．01，差异有显著性。结论在人骨髓基质细胞向成骨细胞分化过程中，骨保护素表达逐渐升高而骨保护素配体表达显著降低，两者比值的逐渐增大，从而发挥促进骨形成，抑制骨吸收的作用，这可能是协调骨重建周期有序进行的重要机制之一。     

Pulsed electromagnetic fields stimulation affects osteoclast formation by modulation of osteoprotegerin, RANK ligand and macrophage colony-stimulating factor

Electromagnetic stimulation has been documented to treat recalcitrant problems of musculoskeletal system. Yet, the underlying mechanisms are not completely understood. In this study, we investigated effect of pulsed electromagnetic fields (PEMF) with parameters modified from clinical bone growth stimulator on osteoclast formation, bone resorption, and cytokines associated with osteoclastogenesis. Marrow cells were harvested from both femora and tibiae of 6 week-old mice and cultured in 8-well chamber slides or 16-well calcium phosphate apatite-coated multitest slides. After 1-day incubation, marrow cells were exposed to PEMF at different electric field intensities for 2h/day and continued for 9 days. Osteoprotegerin (OPG), receptor activator of NFkappaB-ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) concentrations of each group were determined after PEMF stimulation. Osteoclast identity was confirmed by both tartrate resistant acid phosphatase (TRAP) stain and bone resorption assay. A statistically significant increase and decrease of osteoclastogenesis and bone resorption areas were found when exposed to PEMF with different intensities. Besides, consistent correlations among OPG, RANKL, M-CSF, osteoclast numbers, and bone resorption after exposure to different intensities of PEMF were observed. These data demonstrated that PEMF with different intensities could regulate osteoclastogenesis, bone resorption, OPG, RANKL, and M-CSF concentrations in marrow culture system.     

卵巢切除大鼠骨组织白细胞介素-6的基因表达和细胞定位

目的 观察卵巢切除大鼠骨组织白细胞介素－6（IL－6）的基因表达及细胞定位。方法 大鼠的双侧卵巢被切除2个月后,通过RNA斑点杂交直接测骨组织IL－6mRNA的表达,通过原位杂交判断产生IL－6的细胞并进行细胞定位。结果 卵巢切除大鼠骨组织IL－6mRNA表达明显升高,为对照大鼠的2．4倍（P＜0．01）。骨衬里细胞、成骨细胞和骨细胞具有较强的IL－6mRNA杂交信号。结论 骨组织成骨细胞系表达IL－6mRNA,卵巢切除导致IL－6mRNA的表达升高,IL－6在卵巢切除大鼠骨丢失中可能起重要作用。     

Increased RANK ligand in bone marrow of orchiectomized rats and prevention of their bone loss by the RANK ligand inhibitor osteoprotegerin

Orchiectomized (ORX) rats were used to examine the extent to which their increased bone resorption and decreased bone density might relate to increases in RANKL, an essential cytokine for bone resorption.Serum testosterone declined by > 95% in ORX rats 1 and 2 weeks after surgery (p < 0.05 versus sham controls), with no observed changes in serum RANKL. In contrast, RANKL in bone marrow plasma and bone marrow cell extracts was significantly increased (by  100%) 1 and 2 weeks after ORX. Regression analyses of ORX and sham controls revealed a significant inverse correlation between testosterone and RANKL levels measured in marrow cell extracts (R = − 0.58), while marrow plasma RANKL correlated positively with marrow plasma TRACP-5b, an osteoclast marker (R = 0.63). The effects of RANKL inhibition were then studied by treating ORX rats for 6 weeks with OPG-Fc (10 mg/kg, twice/week SC) or with PBS, beginning immediately after surgery. Sham controls were treated with PBS. Vehicle-treated ORX rats showed significant deficits in BMD of the femur/tibia and lower trabecular bone volume in the distal femur (p < 0.05 versus sham). OPG-Fc treatment of ORX rats increased femur/tibia BMD and trabecular bone volume to levels that significantly exceeded values for ORX or sham controls. OPG-Fc reduced trabecular osteoclast surfaces in ORX rats by 99%, and OPG-Fc also prevented ORX-related increases in endocortical eroded surface and ORX-related reductions in periosteal bone formation rate. Micro-CT of lumbar vertebrae from OPG-Fc-treated ORX rats demonstrated significantly greater cortical and trabecular bone volume and density versus ORX-vehicle controls. In summary, ORX rats exhibited increased RANKL protein in bone marrow plasma and in bone marrow cells, with no changes in serum RANKL. Data from regression analyses were consistent with a potential role for testosterone in suppressing RANKL production in bone marrow, and also suggested that soluble RANKL in bone marrow might promote bone resorption. RANKL inhibition prevented ORX-related deficits in trabecular BMD, trabecular architecture, and periosteal bone formation while increasing cortical and trabecular bone volume and density. These results support the investigation of RANKL inhibition as a strategy for preventing bone loss associated with androgen ablation or deficiency.     

Effects of estradiol and dihydrotestosterone on osteoblast gene expression in osteopenic ovariectomized rats

Because androgens increase bone formation and estrogens inhibit bone resorption, there is a potential therapeutic use for a combined treatment of these hormones to preserve bone. We investigated the effect of dihy-drotestosterone (DHT) and estradiol, alone and in combination, on the mRNA levels of genes expressed during osteoblast development in osteopenic ovariectomized (ovx) rats: 40 animals were ovx and administered vehicle or 80mg/kg body weight DHT at 15 weeks postovariectomy. At 19 weeks postovariectomy, the rats were administered vehicle or 20mg/kg body weight estradiol for 1 week. The treatment groups were as follows: (1) vehicle + vehicle, (2) DHT + vehicle, (3) vehicle + estradiol, and (4) DHT + estradiol. Fasting blood and urine samples were collected at 15, 17, 19, and 20 weeks postovariectomy for bone biochemical analyses. On completion of both procedures, the long bones were removed and total RNA extracted. Combined DHT and estradiol treatment increased the mRNA (P < 0.001) and serum levels of alkaline phosphatase (ALP) (P < 0.01) compared to control rats. These data suggest that combined DHT and estradiol treatment stimulates osteoblasts at an early stage of their development when ALP is expressed.     

电针对去卵巢大鼠骨形成及骨吸收活性的影响

目的研究电针刺对去卵巢骨质疏松模型大鼠的全身骨密度(BMD)、骨矿物含量(BMC)及其血清对体外培养成骨细胞的骨形成因子骨钙素(OPG)和骨吸收因子(RANKL)蛋白表达的影响。方法选取SPF级2月龄SD雌性大鼠40只,随机分为空白组、电针组、药物组和模型组,每组10只。其中空白组不做任何处理,后3组采用手术方法切除双侧卵巢。饲养3个月后,骨密度检测结果显示造模成功。随后药物组大鼠灌胃服用戊酸雌二醇3个疗程,电针组大鼠采用针刺后通电治疗3个疗程并服用药物组相同体积的蒸馏水,模型组服用相应体积的蒸馏水,空白组常规饲养。各组大鼠于SPF级实验室处理3个月后,双能X射线骨密度仪检测全身骨密度和骨矿含量。随后,水合氯醛麻醉各组大鼠,心脏采血,各组血液分别经滤网过滤、56℃水浴处理后加入体外培养的大鼠成骨细胞培养液中,处理3d后多聚甲醛固定成骨细胞,采用免疫细胞化学染色法检测成骨细胞内OPG、RANKL蛋白的表达量。结果与空白组相比,模型组大鼠BMD,BMC显著降低(P0.05),服用药物或针刺处理后,BMD和BMC显著增高(P0.05),与模型组相比差异具有统计学意义。各组大鼠血清处理成骨细胞后,模型组骨形成指标OPG蛋白表达量较低,经药物或针刺后,OPG蛋白表达量显著升高(P0.01)。此外,模型组骨吸收指标RANKL的表达量较高,经药物和针刺处理后,RANKL蛋白表达量显著下降(P0.01)。结论针刺能有效升高大鼠BMD和BMC,同时针刺处理后的大鼠血清具有很好的促进骨形成和抑制骨吸收的能力。     

虾青素对去卵巢糖尿病大鼠骨质流失的防治作用

目的探讨虾青素能否防治去卵巢糖尿病大鼠的骨质流失以及可能的机制。方法 3月龄雌性SD大鼠分为3组(每组6只):对照组CON(假手术),模型组OVX/T1DM(去卵巢糖尿病大鼠),药物组OVX/T1DM-ASX(去卵巢糖尿病大鼠,给予虾青素100 mg/(kg·d)。结果连续治疗60 d后,与OVX/T1DM组相比,OVX/T1DM-ASX组骨密度(BMD)明显升高(P0.01),血清I型胶原蛋白(CTX-1)、骨钙素、I型前胶原蛋白n端前肽(PINP)、抗酒石酸磷酸酶5b(TRACP 5b)水平均显著升高(P0.01)。虾青素治疗能抑制去卵巢糖尿病大鼠骨组织形态学的改变,减少骨髓脂肪细胞增加,提高OPG/RANKL的比值。结论虾青素对绝经后糖尿病骨质流失有保护作用,这种作用与调控OPG/RANKL轴有关。     

Changes in serum levels of receptor activator of nuclear factor-kappaB ligand, osteoprotegerin, IL-6 and TNF-alpha in patients with a concomitant head injury and fracture

Introduction  Several reports indicated that interleukin-6 (IL-6) and tumor necrosis factor α (TNF- α) play important regulatory roles in bone remodeling and homeostasis. In addition, receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) have been shown to be important regulators of osteoclastogenesis during bone remodeling, and their expressions were examined during fracture healing in a mouse model of tibial fracture. However, studies linking RANKL, OPG, IL-6 and TNF-α in patients with head injury and fracture are lacking. Patients and methods  Within the first few hours of admission to hospital and at 4, 8, and 12 weeks after the injury, we evaluated changes in serum levels of RANKL, OPG, IL-6 and TNF-α in 24 male patients with a concomitant head injury and fracture and in 26 male patients with fracture only. These levels were compared with those found in 36 healthy controls. Results  The RANKL/OPG ratios were found to significantly lower in patients with a concomitant head injury and fracture than in the controls immediately after admission and at 4, 8, and 12 weeks after the injury. In addition, RANKL/OPG ratios were significantly lower in patients with a concomitant head injury and fracture than in those with fracture at 8 and 12 weeks after the injury. The serum IL-6 levels were significantly higher in patients with a concomitant head injury and fracture than in the controls upon admission, and at 4, 8, and 12 weeks after the injury. Moreover, the serum IL-6 levels were significantly higher in patients with a head injury and fracture than in those with just a fracture at 4, 8, and 12 weeks after the injury. Conclusions  Based on these changes in the profiles of RANKL, OPG, and IL-6 and the RANKL/OPG ratio, altered repair of a fracture can occur in patients with a concomitant head injury and fracture. J. S. Lee and C. H. Ryu contributed equally to this study.     

橙皮苷对去卵巢骨质疏松症大鼠骨质流失和机制的影响

目的研究橙皮苷对去卵巢骨质疏松大鼠骨质流失和骨保护素(osteoprotegerin,OPG)/核因子kappa B配体的受体激活物(receptor activator of nuclear factor kappa B ligand,RANKL)/RANK通路的影响。方法将48只雌性Sprague-Dawley(SD)大鼠随机分为假手术组、去卵巢骨质疏松组、橙皮苷低剂量组、橙皮苷中剂量、橙皮苷高剂量组和雌激素组。检测骨组织骨体积分数(bone volume fraction,BV/TV)、骨小梁厚度(trabecular thickness,Tb.Th)、骨密度(bone mineral density,BMD)、骨小梁数量(trabecular number,Tb.N)、骨小梁分离度(trabecular separation,Tb.Sp),通过HE染色观察骨组织病理损伤,采用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测人Ⅰ型胶原C端肽(C-terminal peptide collagen typeⅠ,CTX-Ⅰ)和血清骨钙素(bone glaprotein,BGP)的表达量。运用qRT-PCR和Western bolt分别检测骨组织OPG、RANKL、RANK的mRNA和蛋白表达。结果在橙皮苷治疗后,与去卵巢骨质疏松组相比,骨质疏松标志物BV/TV、BMD、Tb.Th和Tb.N水平升高,Tb.Sp水平降低;大鼠骨小梁数量明显增加,且骨小梁连接更为紧密;骨吸收标志物CTX-Ⅰ水平降低;骨形成标志物BGP水平升高。OPG mRNA相对表达量和蛋白的表达量升高,RANKL和RANK mRNA相对表达量和蛋白的表达量降低。结论橙皮苷能缓解去卵巢骨质疏松大鼠骨质流失并调节OPG/RANKL/RANK信号通路。     

跑台运动对去卵巢大鼠骨组织中OPG、RANKL和RUNX2 mRNA表达的影响

目的：观察去卵巢大鼠经跑台运动后其骨组织中OPG、RANKL和RUNX2mRNA表达变化,探讨力学刺激防治绝经后骨质疏松的作用机制。方法：健康3月龄雌性SD大鼠30只,体重（270±10）g,按随机方法平均分为假手术组、去卵巢组和跑台运动组,每组10只。假手术组仅在术中牵动卵巢并不行卵巢切除,其余两组均行卵巢切除术。假手术组和去卵巢组大鼠正常饲养;跑台运动组大鼠在术后1周开始在动物跑台上运动,跑速为18m／min,每次45min,1次／d,6d／周,共训练11周。12周后处死所有动物,取左侧股骨头,脱钙后石腊切片,用倒置相差显微镜观察其组织学变化;取右股骨头提总RNA,实时PCR法检测OPG、RANKL和RUNX2的mRNA表达情况。结果：去卵巢组骨小梁稀疏,骨细胞少,跑台运动组骨小梁粗壮增厚。去卵巢组骨组织中OPG的mRNA表达为（0．131±0．080）,RNAKL的mRNA表达为（8．013±3．550）,RUNX2的mRNA表达为（3．245±5．090）。跑台运动组其骨组织中OPG的mRNA表达为（0．566±0．260）,RANKL的mRNA表达为（5．232±3．670）,RUNX2的mRNA表达为（2．753±3．680）。和去卵巢组比较,跑台运动组的OPG的mRNA表达升高,差异有统计学意义;而RANKL和RUNX2的表达下降,差异无统计学意义。结论：力学刺激能通过促进去卵巢大鼠骨组织中OPG的表达而发挥其抗绝经后骨质疏松的效果,这对今后研究绝经后骨质疏松症的治疗提供了新的思路。     

骨保护素体外抑制小鼠单核巨噬细胞 成熟分化的实验研究

目的研究骨保护素（Osteoprotegerin, 0PG)抑制核因子NF-KB受体活化因子配体（Receptor activator of nuclear kappa B ligand,RANKL)诱导小鼠单核细胞RAW264. 7成熟分化而导致的溶骨效 应。方法50 ng/mL RANKL诱导RAW264. 7细胞1 d后,加人100 ng/mL 0PG(实验组,即0PG + RANKL组）或不加人0PG(对照组,即RANKL组）分别培养7 d和9 d,经细胞形态学观察其变化,抗 酒石酸酸性碟酸酶（Tartrate resistant acid phosphatase, TRAP)染色法观察TRAP阳性多核细胞,扫描 电镜下观察在骨片上的破骨细胞所致的骨吸收陷窝形成情况。结果对照组培养7 d时,在倒置相 差显微镜、透射电镜、光镜下可见细胞形状为椭圆形或不规则形,胞体明显较KAW264.7细胞增大, 胞核多为6 ~ 10个,扫描电镜下还可见大量伪足形成,而实验组培养7 d后,细胞形状多为圆形,且扫 描电镜下未见明显伪足形成;对照组9 d时可见大量TRAP染色阳性的多核巨细胞（含3个或3个以 上的细胞核）,而实验组中TRAP染色阳性的多核破骨细胞偶见多核巨细胞,培养9 d时很难找到多 核巨细胞;仅用RANKL诱导RAW264.7细胞分化7 d时,对照组中破骨细胞表面可见大量伪足伸出, 并形成明显的骨吸收陷窝,实验组中破骨细胞见少许伪足突出,不能看到明显的骨陷窝形成。结论 单用50 ng/mL RANKL体外连续诱导RAWM4.7细胞7 d时,可以促进成熟的破骨细胞显著分化。 100 ng/mL 0PG培养9 d能有效地抑制破骨细胞的分化,减少破骨细胞的骨吸收效应。