强骨康疏胶囊对去卵巢骨质疏松大鼠骨细微结构及骨代谢的影响

目的:观察强骨康疏胶囊对去卵巢骨质疏松大鼠的骨密度、OPG及RANKL蛋白表达、骨组织形态计量学参数及骨组织细微结构的影响。方法:制备去卵巢骨质疏松大鼠模型后,分组:正常对照组、模型空白组、中药低剂量预防组、中药高剂量预防组、雌激素预防组。给药1月后,检测各组股骨骨密度值,显微镜下观察股骨骨小梁的结构变化,并检测骨组织形态计量学参数。采用免疫组织化学染色法检测大鼠股骨OPG及RANKL蛋白表达。结果:模型空白组大鼠股骨骨密度减少,骨小梁厚度、面积、面积百分数均减少,骨小梁间距增大,股骨OPG蛋白平均光密度值显著降低,RANKL蛋白平均光密度值明显增高;雌激素预防组、中药低剂量组、中药高剂量组对上述指标均有明显改善。结论:强骨康疏胶囊能有效提高骨量,维持骨小梁立体空间结构,改善大鼠股骨远端松质骨的显微结构,能够提高骨OPG蛋白表达及抑制RANKL蛋白表达。     

咀嚼力增强对骨质疏松大鼠颌骨骨丢失的保护作用及机制

为研究咀嚼力增强对骨质疏松大鼠颌骨骨丢失的保护作用及机制,选取30只雌性SD大鼠进行实验,行摘除卵巢术建立骨质疏松动物模型,喂食高硬度饲料建立咀嚼力增强动物模型,检测血清雌二醇(E2)、骨钙素(BGP)、碱性磷酸酶(ALP)、牙槽骨骨形态计量学指标、TGF-β与OPG、RANKL基因等指标。结果显示,与模型组相比,实验组血清E2增加、BGP和ALP降低,但差异不显著。与模型组相比,实验组牙槽骨TGF-β、OPG显著增加,RANKL显著降低,骨体积分数(BV/TV)显著增加。结果表明,咀嚼力增强能够影响骨质疏松大鼠牙槽骨TGF-β与OPG、RANKL等指标的表达,可延缓骨质疏松大鼠颌骨骨丢失,对骨质疏松大鼠颌骨骨丢失有一定保护作用。     

中药独活激活Wnt/β-catenin信号通路对骨质疏松大鼠的作用

目的探讨中药独活对去卵巢所致骨质疏松SD大鼠的作用及其可能机制。方法将50只SD大鼠,体重200～250 g,随机分为假手术组、模型组、独活低剂量组(0.9 g/kg)、独活高剂量组(1.8 g/kg)、阳性药组(200 mg/kg依普黄酮),每组10只。利用双侧卵巢切除法制备大鼠骨质疏松模型,手术模型制备成功后,连续灌胃给药60d。测定股骨密度、骨灰重/干重百分比;测定大鼠血清骨钙素(OC)、骨碱性磷酸酶(ALP)、抗酒石酸酸性磷酸酶(TRAP)、血清钙、无机磷的水平。免疫组化法测定股骨中骨保护素(OPG)、破骨细胞分化因子(RANKL)蛋白表达;Western blot法检测大鼠骨组织中β-catenin、p-β-catenin、GSK-3β、p-GSK-3β蛋白表达。结果独活高剂量和阳性药均能够增加大鼠股骨密度、骨灰重/干重,降低BGP、ALP及TRAP的水平,并增加血清钙的水平,对无机磷的表达无显著性影响,增加骨组织中OPG蛋白表达水平,降低RANKL蛋白表达水平,增加β-catenin、p-GSK-3β蛋白表达水平,降低p-β-catenin、GSK-3β蛋白表达。结论独活高剂量(1.8 mg/kg)对去卵巢导致的骨质疏松大鼠的治疗作用,与激活Wnt/β-catenin信号通路促进骨形成相关。     

高甘油三酯对去卵巢大鼠股骨RANKL/OPG mRNA表达的影响

目的:探讨高甘油三酯(TG)对去卵巢大鼠股骨核因子-κB受体活化因子配体/护骨素(RANKL/OPG)mRNA表达的影响及非诺贝特的作用。方法:将3月龄雌性SD大鼠40只,用果糖饲养复制高TG模型,大鼠分为4组:(1)去卵巢+果糖组;(2)去卵巢+果糖+非诺贝特(FF)组;(3)去卵巢+普食组;(4)假手术+果糖组。12周后取股骨检测RANKL/OPG mRNA表达水平。结果:(1)去卵巢+果糖组的TG水平高于去卵巢+普食组(P0.01),也高于去卵巢+果糖+FF组(P0.01);(2)去卵巢+果糖组RANKL/OPG mRNA水平,均高于去卵巢+果糖+FF组、去卵巢+普食组和假手术+果糖组(P0.01或P0.05)。结论:高果糖饮食可以诱导去卵巢大鼠形成高甘油三酯血症,后者使股骨组织中RANKL/OPG mRNA增加;非诺贝特可降低TG,保持RANKL/OPG mRNA的平衡。     

脂肪间充质干细胞成骨分化条件培养基联合骨形态发生蛋白2治疗去卵巢大鼠骨质疏松症

文题释义： 骨质疏松症：是由多种原因引起的老年性骨病。通常由于骨量下降、骨的细微结构发生改变,导致骨脆性增加,易发生骨折,好发于胸、腰椎椎体、桡骨远端及股骨上端等。骨质疏松可发生在任何年龄和任何性别,但通常多发生于绝经后妇女和老年男性。 RANKL/OPG：骨重建需要骨形成和骨吸收之间的精确平衡,RANKL/OPG在骨重建中起着至关重要的作用。RANKL是一种细胞因子,可诱导祖细胞分化为成熟的破骨细胞。OPG通过与RANKL结合而成为诱骗受体。因此,RANKL/OPG的比例是维持骨形成和骨吸收之间平衡的关键。 背景：脂肪间充质干细胞分泌各种骨重塑需要的细胞因子和生长因子,被认为是骨再生的优良候选细胞。骨形态发生蛋白2与脂肪间充质干细胞对骨再生有协同作用,并能显著增强脂肪间充质干细胞的成骨分化作用。 目的：探讨脂肪间充质干细胞成骨分化条件培养基与骨形态发生蛋白2联合应用对大鼠绝经后骨质疏松症的影响。 方法：8-10月龄雌性SD大鼠75只,随机取60只采用卵巢切除方法建立绝经后骨质疏松症大鼠模型,余15只进行假手术,未切除卵巢。将60只造模成功大鼠随机分为4组：骨质疏松症组、条件培养基组、骨形态发生蛋白2组、联合治疗组,通过尾静脉分别注射DMEM培养基、脂肪间充质干细胞成骨分化条件培养基、骨形态发生蛋白2、脂肪间充质干细胞成骨分化条件培养基联合骨形态发生蛋白2。治疗12周,取各组大鼠股骨和血清,组织学观察骨小梁数量和结构以及骨小梁间距,ELISA检测血清P1NP、ALP、TRAP、OPG、RANKL水平,Western blot、Realtime PCR检测RANKL、OPG蛋白和mRNA水平,细胞因子芯片分析脂肪间充质干细胞成骨分化条件培养基中细胞因子水平。 结果与结论：①与假手术组相比,骨质疏松症组大鼠的骨小梁间距扩大,骨小梁数量明显减少,骨小梁失去正常结构并且不连续。与骨质疏松症组、条件培养基组、骨形态发生蛋白2组比较,联合治疗组大鼠骨小梁间距扩大较少,骨小梁结构更完整、更连续;②与骨质疏松症组、条件培养基组、骨形态发生蛋白2组比较,联合治疗组大鼠血清P1NP和ALP水平显著升高(P < 0.05),血清TRAP水平显著降低(P < 0.05);③与骨质疏松症组、条件培养基组、骨形态发生蛋白2组比较,联合治疗组RANKL/OPG比值显著降低(P < 0.01),从而促进更多的骨形成;④脂肪间充质干细胞成骨分化条件培养基含有多种与骨形成密不可分的细胞因子,包括骨形态发生蛋白4和7、白血病抑制因子、脑源性神经营养因子、骨保护素、胰岛素样生长因子1等;⑤结果表明,脂肪间充质干细胞成骨分化条件培养基与骨形态发生蛋白2联合应用可减轻卵巢切除大鼠骨质疏松,可能成为治疗绝经后骨质疏松症的新方案。 ORCID: 0000-0003-0552-4818(杨九杰) 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

非诺贝特对高甘油三酯血症并骨质疏松大鼠骨代谢的影响

目的探讨非诺贝特对去卵巢骨质疏松并高甘油三酯（TG）血症大鼠股骨中细胞核因子κB受体活化因子配体/护骨素（RANKL/OPG）mRNA表达的影响。方法将40只3月龄雌性SD大鼠,用果糖饲养复制高TG模型,大鼠共分为去卵巢＋果糖组、去卵巢＋果糖＋非诺贝特（FF）组、去卵巢＋普食组、假手术＋果糖组。12周后取股骨检测细胞核因子κB受体活化因子配体（RANKL）mRNA、护骨素（OPG）mRNA表达水平。结果去卵巢＋果糖组的TG水平高于去卵巢＋普食组（P〈0.01）,也高于去卵巢＋果糖＋FF组（P﹤0.01）;去卵巢＋果糖组RANKL mRNA/OPG mRNA水平,均高于去卵巢＋果糖＋FF组、去卵巢＋普食组和假手术＋果糖组（P〈0.01或〈0.05）。结论非诺贝特可通过降低TG而保持RANKLmRNA/OPGmRNA的平衡。     

姜黄素对佐剂性关节炎大鼠滑膜OPG和RANKL蛋白表达的影响

目的:通过观察姜黄素对佐剂性关节炎(Adjuvant arthritis,AA)模型大鼠滑膜病理、滑膜骨保护素(Osteoprotegerin,OPG)、核因子κB受体活化因子配体(Receptor activator for nuclear factor-κB ligand,RANKL)表达的影响,探讨其防治类风湿关节炎(Rheumatoid arthritis,RA)可能作用机制。方法:雄性SD大鼠以弗氏完全佐剂造模,分为模型组与姜黄素组,另有正常组,d 28处死所有大鼠行滑膜常规病理HE染色,并用Western blot法来检测滑膜RANKL、OPG蛋白的表达。结果:模型组大鼠滑膜炎性细胞浸润、纤维细胞及滑膜细胞的增生较正常组均明显增加(P<0.01),而姜黄素组的大鼠滑膜组织病理较模型组显著改善(P<0.01);姜黄素组大鼠的滑膜RANKL表达显著低于模型组,滑膜OPG表达显著高于模型组,RANKL/OPG比值显著低于模型组(P<0.01)。结论:姜黄素不仅改善AA大鼠关节滑膜病理学损伤,还可调节滑膜RANKL/OPG的比值。因此姜黄素可能通过调节OPG/RANKL系统来发挥对AA大鼠的治疗作用。     

补肾通络方抑制NF-κB/RANK/RANKL通路减轻胶原蛋白诱导关节炎(CIA)大鼠骨破坏

目的探讨补肾通络方(BSTL)对胶原蛋白诱导关节炎(CIA)大鼠模型骨破坏的改善作用以及对核因子κB/核因子κB受体激活蛋白/核因子κB受体激活蛋白配体(NF-κB/RANK/RANKL)信号通路的影响。方法将SD大鼠随机分为空白对照组、CIA模型组、1 mg/kg甲氨喋呤(MTX)处理组、(0.5、2) g/kgBSTL处理组,每组10只。除空白对照组外,其余大鼠采用含有卡介苗的完全Freund佐剂和牛Ⅱ型胶原蛋白(Col2)混合乳液免疫刺激,建立CIA模型。建模15 d后(第0天)根据关节炎指数(AI)评价造模是否成功。自第0天起,连续给予MTX或BSTL处理28 d,免疫组织化学染色法检测滑膜组织NF-κB p65的表达,ELISA检测大鼠血清白细胞介素1β(IL-1β)、IL-18、肿瘤坏死因子α(TNF-α)、抗总Col2抗体及亚型(Col2-IgG、Col2-IgG1、Col2-IgG2a)水平,Western blot法检测滑膜组织RANKL、RANK、护骨因子(OPG)蛋白表达。结果与CIA模型组相比,2 g/kg BSTL处理28 d的大鼠足容积、AI值降低,血清IL-1β、IL-18、TNF-α、Col2-IgG、Col2-IgG2a以及RANK和RANKL水平均降低,OPG水平升高;大鼠滑膜组织中NF-κB p65、RANK和RANKL蛋白表达均降低,OPG蛋白表达增加。结论 BSTL可通过抑制全身炎症反应,降低滑膜组织中NF-κB p65、RANK、RANKL蛋白的表达,上调OPG蛋白表达,缓解CIA模型大鼠关节炎症和肿胀。     

淫羊藿苷对大鼠血管平滑肌细胞钙化的抑制作用

目的:研究淫羊藿苷(Icariin)对大鼠血管平滑肌细胞(VSMCs)钙化的抑制作用。方法:组织贴壁法培养大鼠胸主动脉VSMCs,取5-8代细胞分为对照组、钙化模型组和淫羊藿苷干预A、B、C组。对照组用DMEM培养基培养;钙化模型组用含10mmol/Lβ-甘油磷酸盐(β-GP)的DMEM培养基培养;干预A、B、C组均以钙化模型组为基础分别与10-6 mol/L、10-5 mol/L和10-4 mol/L Icariin的DMEM培养基共培养,茜素红-S染色法鉴定各组细胞钙化情况,硝基苯酚法测定各组细胞碱性磷酸酶(ALP)活性,RT-PCR检测各组细胞内骨保护素(OPG)和核因子κB(NF-κB)受体活化因子配体(RANKL)的mRNA水平,用Western blotting检测各组细胞OPG和RANKL的蛋白表达。结果:与对照组比较,钙化模型组VSMCs发生细胞钙化,并形成红色结节,ALP活性显著升高(P0.01),OPG、RANKL mRNA和蛋白表达均增加(P0.01);与钙化模型组比较,干预A、B、C组细胞钙化减少,ALP活性减低(P0.01),OPG、RANKL mRNA和蛋白表达均下调(P0.01),尤以干预B组效果最显著。结论:淫羊藿苷可能通过降低ALP活性及OPG、RANKL表达来抑制动脉钙化。     

彝药恒古骨伤愈合剂缓解去势树鼩骨质疏松症

目的探讨恒古骨伤愈合剂(OK)可否缓解去势树鼩模型的骨质疏松症(OP)。方法雌性树鼩随机分为正常组和OP模型组去除双侧卵巢(经检测全身骨密度下降25%),每组8只。分别给予OK 0.5、1.5和3. 0 mL/d灌胃治疗,阳性药物对照组给予1. 5 mL/d雌激素灌胃治疗,10周后处死树鼩,收集血清和骨骼,检测血清和尿液中Ca~(2+)、P~(3+)的浓度,E2、BGP和ALP的浓度以及BMD;分别用RT-qPCR和Western blot检测OPG、RANKL和RUNX2的表达。结果低、中、高浓度OK可显著增高OP血清中Ca、P、E2和ALP的浓度(P0. 05),降低血清中BGP和尿液中Ca~(2+)、P~(3+)的浓度(P0. 05),上调骨组织中OPG和RUNX2的表达(P0. 05),下调RANKL的表达。结论 OK能够通过增加E2分泌、抑制骨吸收和促进骨形成,增加全身骨密度,治疗OP。     

补肾补骨汤治疗绝经后骨质疏松症的疗效及分子机制

目的:明确补肾补骨汤(BuShenBuGuTang,BSBGT)治疗绝经后骨质疏松症的分子机制。方法:SD成年雌鼠分为阴性对照组、去势组、中药治疗组和雌激素治疗组4组,12周后检测4组骨密度、骨形态计量学参数及I型胶原的变化并比较各组间差异。体外实验采用大鼠骨髓基质细胞的原代培养体系,加用不同药物处理后,用半定量RT-PCR方法检测破骨细胞分化因子和护骨因子的变化情况。结果:补肾补骨汤对SD大鼠上述3项指标的维持具有明显的效果,而且在体外可明显地抑制破骨细胞分化因子的表达和促进护骨因子的表达。结论:补肾补骨汤对于绝经后骨质疏松症有明显的治疗作用,是通过调节破骨细胞分化因子和护骨因子的表达和分泌而发挥作用的。     

Possible antiosteoporotic mechanism of Cicer arietinum extract in ovariectomized rats

Objective: The present study aimed to throw the light on the anti-osteoprotic mechanism of Cicer arietinum extract (CAE) seeds against ovariectomized (OVX) rats. Methods: Seventy female rats were divided into two groups. The first group (14 rats/group) represented normal rats (Sham operated) while the second group (56 rats/group) underwent bilateral ovariectomy (OVX). After one week of recovery from ovariectomy surgery, the second group was randomly subdivided into 4 subgroups (14 rats/ each subgroup). The rats administered orally; distilled water (vehicle) (1^st subgroup), Cicer arietinum extract (CAE) (500 or 1000 mg/kg body weight/day) (2^nd and 3^rd subgroups), alendronate (6.5 mg/kg mg/kg body weight) as a positive control one time/week (4^rh subgroup), daily for 10 weeks. Results: The present study demonstrated that ovariectomy caused significant decrease in bone mineral; density (BMD) and content (BMC), Bone-specific alkaline phosphatase (BALP), calcium (Ca), phosphorus (P), parathyroid hormone (PTH) and calcitonin levels. Furthermore, ovariectomy induced significant elevation of tartrate-resistant acid phosphatase 5b (TRAP 5b) and receptor activator of nuclear factor (NF-kappa β) ligand (RANKL) concentration. Conversely, osteoprotegerin (OPG) and OPG/RANKL ratio were decreased following ovariectomy. The present work suggests that CAE has antiosteoporotic action against ovariectomy effects and its activity may results from its phytochemical and/or phytoestrogen contents. Conclusion: The ongoing study speculates that the CAE exerts its action through regulation of RANK/RANKL/OPG system. As, CAE not only promotes osteoblast differentiation, but also up-regulates OPG and downregulates RANKL secretion in osteoblasts, subsequently prevents bone loss and osteoporosis.     

Curculigo orchioides, a traditional Chinese medicinal plant, prevents bone loss in ovariectomized rats

OBJECTIVE: Natural medicines derived from plants have aroused increasing interest in the prevention and treatment of osteoporosis. This is due to their unique characteristics as these are more suitable for long-term use compared with synthesized chemicals and have apparently fewer adverse effects. Curculigo orchioides (CO) has a long history in the treatment of postmenopausal osteoporosis in traditional Chinese medicine. The present study was designed to investigate the protective effects of ethanol extracts of CO on ovariectomy-induced bone loss. METHODS: Sixty female (4.5-month-old) Sprague-Dawley rats were assigned to sham and OVX groups. The OVX rats were further divided into five subgroups treated respectively, with vehicle, nylestriol (1 mg/kg, i.g.) and CO extract (0.5, 1.0, and 2.0 g/kg, i.g.) for 12 weeks. Bone mineral density (BMD) and bone mineral content (BMC) were measured by peripheral quantitative computerized tomography (pQCT) densitometry. Serum phosphorus, calcium, ACTH, corticosterone, deoxypyridinoline crosslinks to creatinine ratio (DPD/Cr), alkaline phosphate (ALP), tartrate-resistant acid phosphatase (TRAP), osteoprotegerin (OPG), IL-6, and TNF-alpha were also determined. RESULTS: Administration of CO extract prevented bone loss in the trabecular bone of the tibia in ovariectomized rats without affecting the weight of the body and the uterus, and increased serum phosphorus, calcium, and OPG levels, decreased serum DPD/Cr, TRAP, ACTH, and corticosterone levels, but did not alter serum TNF-alpha, IL-6, and ALP levels in ovariectomized rats. CONCLUSION: CO ethanol extract has a definite protective effect on bone loss in ovariectomized rats by inhibiting bone resorption and increasing serum phosphorus and calcium levels, without affecting bone formation. Therefore, CO can be considered a potential antiosteoporosis herbal plant, although more studies are needed to clarify its real potential chemical constituents and their mechanism of action.     

Association between osteoprotegerin (OPG), receptor activator of nuclear factor-kappaB (RANK), and RANK ligand (RANKL) gene polymorphisms and circulating OPG, soluble RANKL levels, and bone mineral density in Korean postmenopausal women

OBJECTIVE: To investigate the association between osteoprotegerin (OPG), receptor activator of nuclear factor-kappaB (RANK), and RANK ligand (RANKL) gene polymorphisms and circulating OPG, soluble RANKL (sRANKL) levels, and bone mineral density (BMD) in Korean postmenopausal women. DESIGN: The OPG gene A163G, G209A, T245G, and G1181C polymorphisms, the RANK gene C421T and C575T polymorphisms, and the RANKL rs12721445 and rs2277438 polymorphisms were analyzed in 385 Korean postmenopausal women. Levels of serum OPG, soluble RANKL, osteocalcin, C-telopeptide of type I collagen, parathyroid hormone, calcium, and phosphorus and BMD at the lumbar spine and femoral neck were measured. RESULTS: The A163G, G209A, and T245G polymorphisms in the OPG gene were in complete linkage. The RANK C421T and C575T polymorphisms and the RANKL rs12711445 polymorphism were not observed. An association with BMD was found only for the OPG G1181C polymorphism, and BMD at the lumbar spine in women with the CC genotype was significantly higher than in women with the GC or GG genotype, with a C allele dose effect. In itself, the RANKL rs2277438 polymorphism was not related to BMD, but by combining the RANKL genotypes with the GC genotypes of the OPG G1181C polymorphism, the association with BMD at the lumbar spine became significant. No significant differences in the levels of any biochemical marker among genotypes of these polymorphisms were found. CONCLUSIONS: The OPG gene G1181C polymorphism, alone and in combination with the RANKL rs2277438 polymorphism, was identified as a genetic factor associated with BMD of the lumbar spine in Korean women.     

低剂量唑来膦酸对去势拔牙大鼠破骨及成骨细胞的影响

文题释义：去势大鼠：即通过去势法建立骨质疏松动物模型,该模型建造方法主要有3种：去势法、维甲酸灌胃法和糖皮质激素肌注法,而去势法是目前最常用、最成熟的造模方法,主要是通过去除大鼠双侧卵巢至少3个月,构建骨量少、骨显微结构退化而引起骨脆性增加及骨折发生率升高的一种全身性骨病,即骨质疏松疾病模型。 TUNEL细胞凋亡检测：用于检测细胞在凋亡过程中细胞核DNA的断裂情况,其原理是生物素标记的dUTP在脱氧核糖核苷酸末端转移酶的作用下,可以连接到凋亡细胞中断裂DNA的3’-OH末端,并可与连接辣根过氧化酶的链霉亲和素特异结合,在显色剂DAB的存在下,产生很强的颜色反应(呈深棕色),特异准确定位正在凋亡的细胞。 背景：目前唑来膦酸虽能有效地预防绝经后妇女的骨丢失,但其对下颌骨的影响及其机制尚不清楚。 目的：观察去势大鼠经低剂量唑来膦酸作用后下颌骨组织病理学改变,探讨RANKL/RANK/OPG信号系统在唑来膦酸抑制骨吸收过程中的调控效应与机制。 方法：取36只雌性SD大鼠随机分为对照组、模型组和治疗组,后2组大鼠行两侧卵巢切除术,建立骨质疏松模型;对照组行假手术去除同等质量卵巢周围的脂肪。去卵巢3个月后,治疗组皮下一次性注射唑来膦酸20 μg/kg,对照组和模型组皮下注射相应剂量的盐水;用药1周后拔除大鼠左侧下颌磨牙,拔牙后4周麻醉动物取出拔牙侧下颌骨组织。通过影像学X 射线大致观察大鼠拔牙窝剩余牙槽骨状况,苏木精-伊红染色检测下颌骨皮质和骨松质的病理结构改变,TUNEL凋亡实验检测凋亡的成骨细胞数量,利用免疫组织化学技术检测下颌牙槽骨内细胞核因子κB受体活化因子配基(RANKL)、骨保护素、细胞核转录因子(NF-κB)的表达情况,最后Western blot检测核因子κB受体活化因子配基、细胞核转录因子蛋白的表达。 结果与结论：①拔牙4周后,治疗组相较模型组牙槽骨骨吸收减少,拔牙窝底有新骨形成;②苏木精-伊红染色可见,模型组的骨皮质变薄,骨小梁结构变细,甚至出现断裂,大量骨吸收陷窝,成骨细胞较少;治疗组骨皮质增厚,骨小梁结构正常,只有少量骨吸收陷窝,成骨细胞增多;③治疗组的成骨细胞凋亡数目显著低于模型组(P < 0.001);④免疫组织化学染色可见,治疗组RANKL蛋白、NF-KB p65蛋白表达显著低于模型组(P < 0.001,P < 0.002),骨保护素蛋白表达显著高于模型组(P < 0.001);⑤Western Blot结果显示,与对照组相比较,模型组的RANKL、NF-κB蛋白高表达,治疗组其表达量显著降低(均P < 0.001);⑥结果说明,唑来膦酸可通过下调细胞核转录因子信号通路抑制破骨细胞的分化,同时调控成骨细胞的凋亡。 ORCID: 0000-0002-0300-0460(程余婷) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

川芎嗪对类风湿性关节炎外周血RANK／RANKL／骨保护素途径干预作用的研究

目的观察川芎嗪对类风湿性关节炎（RA）模型大鼠RANK／RANKL／0PG在外周血中CD3^＋T淋巴细胞、中性粒细胞、CD14单核细胞的表达率及平均荧光强度,探讨川芎嗪在RA骨破坏和炎症过程巾的意义。方法大鼠随机分成5组,正常对照组、模型对照组、川芎嗪大剂量组、川芎嗪小剂量组、阳性药物对照组,每组10只。给药7d后,应用间接免疫荧光标记和流式细胞技术对各组大鼠外周血相关指标进行检测分析。结果与正常对照组相比,RA大鼠骨保护素（OPG）表达明显降低,从正常的24．7降至18．7（q=4．2,P〈0．05）,RANK、RANKL变化不明显,经过川芎嗪治疗后,大剂量组OPG有明显的回升,至23．8％（q=3．97,P〈0．05）,小剂量组变化不明显。RANK在CD3^＋细胞、中性粒细胞、CD14单核细胞上的平均荧光强度明显降低,分别为20．6、135．4、84．2,经川芎嗪大剂量治疗后明显升高,分别达到31．0、192．1、95．6（q=10．4、q=8．6、q=6．3,P〈0．05）。结论大剂量川芎嗪可以通过调节OPG／RANK／RANKL途径对RA起一定的作用。     

Osteoprotegerin/RANK/RANKL axis in cardiac remodeling due to immuno-inflammatory myocardial disease

Objective

Osteoprotegerin (OPG) and the receptor activator of nuclear factor-κB ligand (RANKL) are inflammatory cytokines traditionally linked to the regulation of bone remodeling. We hypothesize that the OPG/RANK/RANKL axis may be involved in extracellular matrix remodeling in immuno-inflammatory heart diseases, and explore the probable underlying mechanisms by using anti-IL-17 in the model of experimental autoimmune myocarditis.

Methods

EAM was induced in Lewis rats by injection of porcine cardiac myosin. All the rats were randomly distributed into day 0, day 7, day 14 and day 28 groups, which means rats in certain group were cervical dislocated on day 0, day 7, day 14 and day 28 respectively. HE staining and Masson's staining were used for measurement of cardiac hypertrophy and interstitial fibrosis. Hydroxyproline content and collagen cross-linking were determined in heart section. Anti-IL-17 or control antibody was injected i.p. 2 h before and 3 days and 7 days after the first myosin immunization in EAM model, that is, group anti-IL-17 or group control antibody. IL-17 and OPG/RANK/RANKL axis expressions were detected by realtime RT-PCR in all the six groups. In the in vitro studies, cardiac fibroblasts were cultured and treated with IL-17 or vehicle for 48 h. Total RNA was isolated from harvested cells and realtime RT-PCR was performed to detect the RANK, RANKL, OPG and MMP-2, MMP-9, TIMP-1 and TIMP-2 expressions, then matrix metalloproteinase activity was assayed.

Results

Our in vivo results revealed that expression of IL-17 and the OPG/RANK/RANKL axis increased significantly from day 0 to day 28, with IL-17 and OPG increased relatively steeply. In the in vitro study, we detected OPG, RANK and RANKL mRNA expressions in the cultured fibroblasts with or without IL-17 stimulation. We found that IL-17 increased the OPG/RANK/RANKL axis activity (P < 0.05). Although IL-17 induced a significant increase in MMP-2 and MMP-9 gene expressions in cardiac fibroblasts, there was no change in TIMP-2 and TIMP-1 expressions.

Conclusions

Our results suggest that the OPG/RANK/RANKL axis may be involved in cardiac remodeling in immuno-inflammatory myocardial diseases and progression of chronic HF and thus may represent targets for intervention in this disorder.     

聚甲基丙烯酸甲酯颗粒对人滑膜细胞核因子κB受体激动剂配体/骨保护蛋白表达的影响

背景：多种因子可以影响骨保护蛋白/核因子κB受体激动剂配体/核因子κB受体激动剂(Osteoprotegerin / receptor activator of nuclear factor-kappaB/ligand of receptor-activator of nuclear factor-kappaB,OPG/RANKL/RANK)系统的表达影响骨代谢,那么松动假体周围的骨水泥颗粒是否也通过影响其代谢参与假体松动过程？ 目的：观察聚甲基丙烯酸甲酯颗粒对人滑膜细胞RANKL/OPG表达的影响。 方法：体外培养人滑膜细胞,在滑膜细胞培养体系中分别加入质量浓度为0(空白对照),0.2,1,10 g/L的聚甲基丙烯酸甲酯骨水泥颗粒,采用荧光定量PCR检测上述各浓度PMMA颗粒作用不同时间后滑膜细胞内RANKL、OPG的表达量及其比值。 结果与结论：与空白对照组相比,各实验组滑膜细胞内RANKL、OPG的表达量均有下降;随着与聚甲基丙烯酸甲酯颗粒共培养时间延长,各实验组滑膜细胞内RANKL、OPG表达均有下降趋势,而RANKL/OPG表达比值无明显变化。说明聚甲基丙烯酸甲酯颗粒在对滑膜细胞产生生物学反应时并不干扰假体周围骨代谢的动态平衡,并未直接参与人工关节假体的无菌性松动。