Role of osteoprotegerin/receptor activator of nuclear factor kappa B/receptor activator of nuclear factor kappa B ligand axis in nonalcoholic fatty liver disease

Concomitantly with the increase in the prevalences of overweight/obesity, nonalcoholic fatty liver disease(NAFLD) has worldwide become the main cause of chronic liver disease in both adults and children. Patients with fatty liver display features of metabolic syndrome(Met S), like insulin resistance(IR), glucose intolerance, hypertension and dyslipidemia. Recently, epidemiological studies have linked obesity, Met S, and NAFLD to decreased bone mineral density and osteoporosis, highlighting an intricate interplay among bone, adipose tissue, and liver. Osteoprotegerin(OPG), an important symbol of the receptor activator of nuclear factor-B ligand/receptor activator of nuclear factor kappa B/OPG system activation, typically considered for its role in bone metabolism, may also play critical roles in the initiation and perpetuation of obesityrelated comorbidities. Clinical data have indicated that OPG concentrations are associated with hypertension, left ventricular hypertrophy, vascular calcification, endothelial dysfunction, and severity of liver damage in chronic hepatitis C. Nonetheless, the relationship between circulating OPG and IR as a key feature of Met S as well as between OPG and NAFLD remains uncertain. Thus, the aims of the present review are to provide the existent knowledge on these associations and to discuss briefly the underlying mechanisms linking OPG and NAFLD.

Effect of Triptolide on Expression of Receptor Activator of Nuclear Factor-κB Ligand in Rat Adjuvant Induced Arthritis

The effect of triptolide （TP） on the expression of receptor activator of nuclear factor-κB ligand （RANKL） and osteoprotegerin （OPG） was explored in rat adjuvant induced arthritis （AA）. AA was induced in Wistar rats. Arthritis rats were treated with TP and methotrexate （MTX） at the onset （day 9） of arthritis. On the peak of arthritis （day 24）, the expression of RANKL and OPG protein in the joints and RANKL mRNA in peripheral blood mononuclear cells （PBMC） was detected. TNF-α and IL-1β levels in peripheral blood were determined. Bone erosion scores were also evaluated. The results showed that bone erosion scores in TP and MTX groups were lower than in AA group （.P〈0.01） ; The expression levels of RANKL in the synovium （P〈0.01） and bone （P〈0.05）, and OPG level in synovium （P〈0.05） were lower in TP group than in AA group （P〈0.05）. In TP group, the expression levels of RANKL mRNA and TNF-α, IL-1β in PBMC were lower than in AA group （all P〈0.01）. It was concluded that TP could inhibit rat adjuvant arthritis bone erosion by suppressing the expression of RANKL.

Influence of baicalin on the expression of receptor activator of nuclear factor-κB ligand and osteoprotegerin in human periodontal ligament cells

Objective To study the effect of baicalin on the expression of receptor activator of nuclear factor-κB ligand(RANKL)and osteoprotegerin(OPG)in cultured human periodontal ligament(HPDL)cells.Methods Small interfering RNA(siRNA)eukaryotic expression vector targeted transforming growth factor βⅡ receptor(TGF-β RⅡ)was constructed and transfected into T cells.HPDL cells with T cells transfected with siRNA or not were placed in the culture medium that had been added with lipopolysaccharide(LPS)and baicalin.The obtained solution was divided into six groups according to the components(group Ⅰ:HPDL cells+LPS+T cells transfected with siRNA1+baicalin;group Ⅱ:HPDL cells+LPS+T cells transfected with siRNA1;group Ⅲ:HPDL cells+LPS+T cells+baicalin;group Ⅳ:HPDL cells+LPS+T cells;group Ⅴ:HPDL cells+baicalin;group Ⅵ:HPDL cells)and was cultured for 48 hours.RT-PCR was used to observe the effect of baicalin on the expression of OPG-RANKL in HPDL cells.Results The ratio of RANKL/OPG in group Ⅰ was lower than that in group Ⅱ(P<0.01)and higher than that in group Ⅲ(P<0.01);The ratio of RANKL/OPG in group Ⅲ was lower than that in group Ⅳ(P<0.01);the ratio of RANKL/OPG in group Ⅳ was higher than that in group Ⅵ(P<0.01);the ratio of RANKL/OPG in group Ⅴ was lower than that in group Ⅵ(P<0.05).Conclusion ① Baicalin could decrease the ratio of RANKL/OPG in HPDL cells.② The TGF-β signaling transduction plays an important role in the effect of baicalin on the RANKL/OPG ratio in HPDL cells.③ Baicalin acts not only through TGF-β to regulate RANKL/OPG in HPDL cells,but also through other pathways.

龈沟液RANKL、音猬因子水平对牙缺失Nobel种植体早期稳定性评估价值

目的探讨龈沟液核因子-κB受体活化因子配体(RANKL)、音猬因子水平对牙缺失Nobel种植体早期稳定性评估价值。方法选取2020年6月至2021年12月邯郸市第一医院牙缺失患者128例作为观察组,均为Nobel种植体修复,选取同期体检健康者64例作为对照组。对比两组及观察组术后即刻、术后3个月龈沟液RANKL、音猬因子水平、种植体稳定系数(ISQ)值,多重线性回归模型分析Nobel种植体早期稳定性影响因素,并分析龈沟液RANKL、音猬因子水平对早期稳定性评估价值。结果观察组术后3个月龈沟液RANKL、音猬因子水平低于术后即刻,差异有统计学意义(P<0.05);观察组术后3个月ISQ值高于术后即刻,差异有统计学意义(P<0.05)。术后即刻龈沟液RANKL、音猬因子水平与术后3个月ISQ值呈负相关(P<0.05)。术后即刻及术后3个月龈沟液RANKL、音猬因子水平对应的线性系数差异有统计学意义(P<0.05);建立多重线性回归模型:术后3个月ISQ值=100.968-4.607×术后咬(牙合)痛-2.886×常食坚果-0.254×术后即刻龈沟液RANKL-0.280×术后即刻龈沟液音猬因子,回归模型具有统计学意义(P<0.05),自变量可解释术后3个月ISQ值74.59%的变异量。结论牙缺失患者进行Nobel种植后龈沟液RANKL、音猬因子呈低表达,术后通过检测二者水平可有效判断种植体早期稳定性。

围绝经期女性外周血OSTF1、OPG/RANKL对骨质疏松症的预测价值

目的探讨围绝经期女性外周血破骨细胞刺激因子1(OSTF1)、护骨素(OPG)/核因子-κB受体活化因子配基(RANKL)对骨质疏松症的预测价值。方法选择2021年3月至2023年1月河北省妇幼保健中心收治的165例围绝经期骨质疏松症患者为疾病组,并纳入同期120例围绝经期骨密度正常的健康志愿者为对照组。采用Pearson相关分析外周血OSTF1、OPG/RANKL与骨密度的相关性,并收集基线资料,采用单因素分析和多因素Logistic回归分析骨质疏松症发生的影响因素,并使用受试者工作特征(ROC)曲线分析外周血OSTF1、OPG/RANKL对骨质疏松症的预测价值。结果疾病组患者血清OSTF1、RANKL水平明显高于对照组,血清OPG水平、OPG/RANKL明显低于对照组,差异均有统计学意义(P<0.05)。疾病组患者整体骨密度、腰椎骨密度和T值明显低于对照组(P<0.05)。多因素Logistic回归分析结果显示,OSTF1、OPG/RANKL、OPG、整体骨密度、腰椎骨密度、T值和RANKL是骨质疏松症发生的影响因素(P<0.05)。ROC曲线分析结果显示:血清OSTF1预测骨质疏松症发生的曲线下面积(AUC)为0.772(95%CI:0.705~0.843);OPG/RANKL的AUC为0.616(95%CI:0.531~0.699);2项联合的AUC为0.906(95%CI:0.864~0.952)。结论围绝经期女性外周血OSTF1、OPG/RANKL异常表达,OSTF1和OPG/RANKL可应用于骨质疏松症预测,值得临床进一步研究并推广。

Imbalance of osteoprotegerin/receptor activator of nuclear factor-κB ligand and oxidative stress in patients with obstructive sleep apnea-hypopnea syndrome

Background:Obstructive sleep apnea-hypopnea syndrome (OSAHS) is associated with a higher prevalence of osteoporosis.However,the underlying mechanisms linking OSAHS with bone loss are still unclear.The aim of this study was to investigate the changes of receptor activator of nuclear factor-κB ligand (RANKL,an osteoclastogenesis-promoting factor) and osteoprotegerin (OPG,the decoy receptor for RANKL),oxidative stress and bone metabolism markers in OSAHS,in order to understand the potential mechanisms underlying bone loss in OSAHS patients.Methods:Forty-eight male patients with OSAHS,confirmed by polysomnography (PSG) study,were enrolled.Twenty male subjects who were confirmed as not having OSAHS served as the controls.The subjects’bone mineral density (BMD) was assessed in lumbar spine and femoral neck using dual-energy X-ray absorptiometry (DXA).Blood samples were collected from all subjects for measurement of RANKL,OPG,the bone formation marker bone-specific alkaline phosphatase (BAP),the bone resorption marker tartrate-resistant acid phosphatase 5b (TRAP-5b),and total antioxidant capacity (TAOC).Results:The BMD and the T-score of the femoral neck and the lumbar spine were significantly lower in OSAHS patients as compared to the control group (P< 0.05).The serum level of BAP was significantly decreased in the OSAHS group (15.62 ± 5.20 μg/L) as compared to the control group (18.83 ± 5.50 μg/L,t= -2.235,P< 0.05),while the levels of TRAP-5b did not differ between the two groups (t= -1.447,P> 0.05).The serum level of OPG and the OPG/RANKL ratio were lower in the OSAHS group compared to the control group (bothP< 0.05).TAOC level was also decreased significantly in the OSAHS group (P< 0.05).Correlation analysis showed that the TAOC level was positively correlated with BAP in the OSAHS group (r= 0.248,P= 0.04),but there were no correlations between TAOC and the BMD or the T-scores.The correlations between the level of OPG (or the OPG/RANKL ratio) and BMD or TAOC did not reach significance.Conclusion:In OSAHS patients,lower levels of TAOC were associated with decreased bone formation,suggesting a role of oxidative stress in bone loss,while the role of OPG/RANKL imbalance in bone metabolism in OSAHS needs further evaluation .

Cannabinoid receptor-2 selective antagonist negatively regulates receptor activator of nuclear factor kappa B ligand mediated osteoclastogenesis

Background The cannabinoid receptor-2 （CB2） is important for bone remodeling. In this study, we investigated the effects of CB2 selective antagonist （AM630） on receptor activator of nuclear factor kappa B （RANK） ligand （RANKL）induced osteoclast differentiation and the underlying signaling pathway using a monocyte-macrophage cell line-RAW264.7.Methods RAW264.7 was cultured with RANKL for 6 days and then treated with AM630 for 24 hours. Mature osteoclasts were measured by tartrate-resistant acid phosphatase （TRAP） staining using a commercial kit. Total ribonucleic acid （RNA）was isolated and real-time reverse transcriptase-polymerase chain reaction （RT-PCR） was done to examine the expression of RANK, cathepsin K （CPK） and nuclear factor kappa B （NF-κB）. The extracellular signal-regulated kinase （ERK）,phosphorylation of ERK （P-ERK） and NF-κB production were tested by Western blotting. The effect of AM630 on RAW264.7 viability was determined using the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazoliumbromide （MTT） assay.Results AM630 did not affect the viability of RAW264.7. However, this CB2 selective antagonist markedly inhibited osteoclast formation and the inhibition rate was dose-dependent. The dose of 〉100 nmol/L could reduce TRAP positive cells to the levels that were significantly lower than the control. AM630 suppressed the expression of genes associated with osteoclast differentiation and activation, such as RANK and CPK. An analysis of a signaling pathway showed that AM630 inhibited the RANKL-induced activation of ERK, but not NF-κB.Conclusion AM630 could inhibit the osteoclastogenesis from RAW264.7 induced with RANKL.

Receptor activator of nuclear factor kappa B ligand and osteoprotegerin expression in chronic apical periodontitis：possible association with inflammatory cells

Background Receptor activator of nuclear factor kappa B （NF-κB） ligand （RANKL） and osteoprotegerin （OPG） have been recently shown to play important roles in bone resorption. The aim of this study was to investigate the possible association between the expression of bone resorption regulators （RANKL and OPG） and inflammatory cell infiltration in chronic apical periodontitis.Methods The samples of chronic periapical lesions （n=40） and healthy periapical tissues （n=10） were examined for immunohistochemical analysis of RANKL and OPG. Lesion samples were further analyzed for the inflammatory infiltration condition. The inflammatory cell infiltration was scored in relation to immunohistochemical reactivity for CD3, CD20 and CD68.Results The number of RANKL-positive cells and the ratio of RANKL/OPG in chronic apical periodontitis were significantly higher than those in healthy periapical tissues （P＜0.001）. The number of RANKL-positive cells was higher in lesions with severe inflammatory infiltration than in those with light inflammatory infiltration （P＜0.05）. Significantly increased RANKL expression was found with T lymphocytes （CD3＋）, macrophages （CD68＋） and B lymphocytes （CD20＋）infiltration （P＜0.05）. No association was found between the ratio of RANKL/OPG and inflammatory cell infiltration.Conclusions RANKL expression was increased with T, B lymphocytes and macrophages infiltration, respectively in chronic periapical lesions. RANKL appears to be closely related to periapical inflammatory infiltrates. The relative ratio of RANKL/OPG may be a key determinant of RANKL-mediated bone resorption.

Effect of Wenhua Juanbi Recipe(温化蠲痹方) on Expression of Receptor Activator of Nuclear Factor Kappa B Ligand,Osteoprotegerin,and Tumor Necrosis Factor Receptor Superfamily Member 14 in Rats with Collagen-Induced Arthritis

Objective： To study the effect of Wenhua Juanbi Recipe（温化蠲痹方, WJR） on expression of receptor activator of nuclear factor kappa B ligand（RANKL）, osteoprotegerin（OPG）, and tumor necrosis factor receptor superfamily member 14（TNFRSF14, also known as LIGHT） in rats with collagen-induced arthritis（CIA）. Methods： CIA rats were generated by subcutaneous injection of bovine collagen type-Ⅱ at the tail base. Sixty CIA rats were randomly assigned（10 animals/group） to： model, methotrexate（MTX）-treated（0.78 mg/kg body weight）, and WJR-treated（22.9 g/kg） groups. Healthy normal rats（n=10） were used as the normal control. Treatments or saline were administered once daily by oral gavage. Rats were sacrificed at day 28 post-treatment and knee synovium and peripheral blood serum were collected. Toe swelling degree and expression of RANKL, OPG, and LIGHT were determined by Western blot and immunohistochemistry. Results： Compared with the normal group, toe swelling degree was significantly increased in the model group（P〈0.01）. After treatment, toe swelling degree decreased significantly in the WJR and MTX groups compared with the model group（P〈0.01）. Compared with the normal group, expression of RANKL and LIGHT were significantly increased and OPG significantly decreased in peripheral blood and synovium of the model group（P〈0.01）. Conversely, RANKL and LIGHT expression were significantly reduced and OPG increased in the WJR and MTX groups compared with the model group（P〈0.01）. No statistically significant difference existed between WJR and MTX groups. Conclusion： WJR likely acts by reducing RANKL expression and increasing OPG expression, thus inhibiting RANKL/RANK interaction and reducing LIGHT expression, thereby inhibiting osteoclast formation/activation to block bone erosion.

辐射对RANKL诱导RAW264.7细胞向破骨细胞分化中CBFα1表达水平的影响

为了研究辐射对于破骨细胞中Notch通路的影响,进一步了解辐射诱发骨损伤的机理,本研究采用核因子κB受体活化因子配体(Receptor activator of nuclear Factor-B Ligand,RANKL)诱导RAW264.7细胞系生成的破骨细胞,经2 Gy 137Csγ射线照射后应用实时定量聚合酶链反应(Real-time polymerase chain reaction,Real-time PCR)方法,检测其Notch通路重要靶位启动子C结合因子α1(C Promoter-Binding Factorα1,CBFα1)表达的变化情况。实验结果表明,在2 Gy 137Csγ射线照射后,CBFα1在RANKL作用下生成的破骨细胞中表达明显增高。辐射诱发的Notch通路中CBFα1表达增高,可能是由于辐射增强了Notch通路对破骨细胞分化的促进作用,增加了破骨细胞的数量,增强了其活性,从而导致骨损伤、骨质疏松、甚至骨折的发病率增高。

RANKL/RNAK/OPG信号通路影响小鼠膝关节假体无菌性松动的作用机制

目的研究RANKL/RANK/OPG信号通路在小鼠膝关节假体无菌性松动中的作用机制。方法28只小鼠以计算机随机数字表法分成正常组、实验组,最终均分别纳入12只。实验组通过在胫骨近端骨髓腔中注射钴铬颗粒悬液建立小鼠膝关节假体无菌性松动模型,正常组注射等量等渗盐水。采用实时荧光定量PCR(qRT-PCR)法检测界膜组织中RANKL、RNAK、OPG mRNA表达量;免疫组化染色法检测界膜组织中RANKL、RNAK、OPG、抗酒石酸酸性磷酸酶(TRAP)蛋白表达量。结果与正常组比较,实验组建模后6、12周界膜组织RANKL、RNAK mRNA及蛋白表达量,RANKL/OPG升高,OPG mRNA及蛋白表达量降低(P<0.05)。与建模后6周比较,正常组建模后12周界膜组织RANKL、RNAK mRNA及蛋白表达量及RANKL/OPG降低,OPG mRNA及蛋白表达量升高(P<0.05),实验组界膜组织RANKL、RNAK mRNA及蛋白表达量及RANKL/OPG升高,OPG mRNA及蛋白表达量降低(P<0.05)。与正常组比较,实验组建模后6、12周界膜组织TRAP蛋白表达量升高(P<0.05)。与建模后6周比较,正常组建模后12周界膜组织TRAP蛋白表达量降低(P<0.05),实验组界膜组织TRAP蛋白表达量升高(P<0.05)。结论小鼠膝关节假体无菌性松动的界膜组织中RANKL、RNAK表达量及RANKL/OPG升高,OPG表达量降低,RANKL/RNAK/OPG信号通路表达失衡与其发生、发展相关。

正畸保持期龈沟液骨保护素/核因子kappa B受体活化因子配体水平对牙槽骨改建状态的意义

目的:探讨正畸保持期牙周龈沟液中骨保护素(osteoprotegerin,OPG)和核因子kappa B受体活化因子配体水平(receptor activator of nuclear factor kappa B ligand,RANKL)的变化及相应牙周组织OPG和RANKL表达的变化,为预测保持期牙周骨组织状态是否稳定提供依据。方法:选择6周龄雄性Wista大鼠15只,按时间点(T1:基线,T2:加力14 d,T3:停止加力14 d)分为3组,每组5只。在每个时间点先采集龈沟液样本,应用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测OPG和sRANKL(soluble receptor activator of nuclear factor kappaB ligand)浓度,然后处死大鼠,采用免疫组织化学检测牙周组织中OPG和RANKL表达。结果:龈沟液中sRANKL浓度在T2期明显上升(P<0.05),T3期则显著下降(P<0.05),与T1期几乎相当。龈沟液中OPG浓度在3个时间点的变化差异均无统计学意义(P>0.05);龈沟液中sRANKL/OPG的比值和牙周组织RANKL/OPG的比值变化近似,均在T2期明显增大(P<0.05和P<0.01),在T3期显著减小(P<0.05)。结论:在正畸加力至保持期,龈沟液中sRANKL/OPG的比值可以在一定程度上反映牙周组织中骨改建的状态。

抗MCV抗体与RANKL、OPG、TRACP-5b及RA疾病活动度的相关性研究

目的探索类风湿关节炎(rheumatoid arthritis,RA)患者抗突变型瓜氨酸波形蛋白(mutant citrulline vimentin,MCV)抗体与骨代谢标志物及疾病活动度之间的关系。方法收集119例RA患者临床资料和血清,检测抗MCV抗体及RANKL、OPG和TRACP-5b等骨代谢标志物,分析抗MCV抗体与骨代谢标志物之间的相关性,并探索抗MCV抗体与疾病活动是否有关联。结果RA患者抗MCV抗体滴度与RANKL、TRACP-5b水平及RANKL/OPG均无显著相关性(P>0.05),与OPG呈正相关但相关系数低(r=0.183,P<0.05)。②抗MCV抗体与DAS28(r=0.376,P<0.01)、ESR(r=0.440,P<0.01)、RF-IgM(r=0.376,P<0.01)呈正相关。结论抗MCV抗体与血清中骨代谢相关标志物TRACP-5b、RANKL、OPG、RANKL/OPG均无显著相关性,提示抗MCV抗体可能不直接通过影响骨代谢参与RA骨侵蚀进展。抗MCV抗体与疾病活动度呈正相关,提示它对RA病情评估可能有一定的价值。

RANKL,a necessary chance for clinical application to osteoporosis and cancer-related bone diseases

Osteoporosis is a common bone disease characterized by reduced bone and increased risk of fracture. In postmenopausal women, osteoporosis results from bone loss attributable to estrogen deficiency. Osteoclast differentiation and activation is mediated by receptor activator of nuclear factor-κB ligand(RANKL), its receptor receptor activator of nuclear factor-κB(RANK), and a decoy receptor for RANKL, osteoprotegerin(OPG). The OPG/RANKL/RANK system plays a pivotal role in osteoclast biology. Currently, a fully human antiRANKL monoclonal antibody named denosumab is being clinically used for the treatment of osteoporosis and cancer-related bone disorders. This review describes recent advances in RANKL-related research, a story from bench to bedside. First, the discovery of the key factors, OPG/RANKL/RANK, revealed the molecular mechanism of osteoclastogenesis. Second, we established three animal models:(1) a novel and rapid bone loss model by administration of glutathione-S transferase-RANKL fusion protein to mice;(2) a novel mouse model of hypercalcemia with anorexia by overexpression of soluble RANKL using an adenovirus vector; and(3) a novel mouse model of osteopetrosis by administration of a denosumab-like anti-mouse RANKL neutralizing monoclonal antibody. Lastly, anti-human RANKL monoclonal antibody has been successfully applied to the treatment of osteoporosis and cancer-related bone disorders in many countries. This is a real example of applying basic science to clinical practice.

Gen对类风湿关节炎RA患者BM-MSCs中RANK/RANKL/OPG系统的影响

目的研究金雀异黄素(Gen)对类风湿关节炎(RA)患者间充质干细胞(MSCs)中的核因子κB受体活化因子(RANK)、核因子κB受体活化因子配体(RANKL)和骨保护素(OPG)系统的影响及其机制。方法分离培养正常人与RA患者的骨髓MSCs(BM-MSCs),分为空白对照组、Gen处理组、雌二醇(E_2)处理组和Gen+雌激素受体拮抗剂(ICI182780)组,采用Western blot法检测BM-MSCs中RANK、RANKL、OPG的表达;采用免疫荧光法检测雌激素受体α(ERα)细胞内分布情况。结果 Western blot结果显示:与正常人BM-MSCs相比,RA患者BM-MSCs中的RANK的蛋白表达水平明显升高,OPG的蛋白表达水平明显降低,RANKL/OPG比值升高;Gen显著上调RA患者BM-MSCs中OPG表达,对RANK、RANKL表达无明显影响,从而降低了RANKL/OPG比值,与E_2作用类似。免疫荧光结果显示:Gen能促进RA患者的BM-MSCs中ERα入核。结论Gen可能通过活化雌激素受体介导的相关信号通路,调控RA患者BM-MSCs中RANKL/OPG的表达,从而调节破骨细胞分化及成骨细胞分化,抑制RA关节破坏和促进受损关节的修复。

Efficacy of recombinant human osteoprotegerin combined with tinidazole in the treatment of periodontitis mice and its correlation with serum RANKL and MCP-1 levels

Objective: To investigate the effect of recombinant human osteoprotegerin combined with tinidazole on mice with periodontitis and the effect on serum RANKL and MCP-1 levels. Methods: 80 SPF-cleaned mice were randomly divided into 4 groups, 20 each, model group, tinidazole group and recombinant human osteoprotegerin group were modeled by Kimura et al., and tinidazole group received tinidazole. After intragastric administration, the recombinant human osteoprotegerin group was injected with recombinant human osteoprotegerin in the periodontal pocket according to the tinidazole group. The periodontal changes of the four groups of mice were observed and recorded, and the gingival rating was performed. Epithelial tissue morphology was observed by hematoxylin-eosin (HE) staining. Serum levels of IL-4, IL-6, RANKL and MCP-1 were measured by enzyme-linked immunosorbent assay. Results:After the intervention, the model group developed severe inflammatory reactions, including redness, hemorrhage, and deep periodontal pockets. The teeth were significantly loosened. The mice in the tinidazole group and the recombinant human osteoprotegerin group recovered substantially, and the gingival rating of the recombinant human osteoprotegerin group was better than that. The tinidazole group and the model group (P<0.05). The results of HE staining showed that the model group had edema, vasodilation and a large amount of inflammatory infiltration. The epithelial structure of the mice in the tinidazole group and the recombinant human osteoprotegerin group was intact and arranged closely and orderly. After intervention, the IL-4 in the tinidazole group and the recombinant human osteoprotegerin group was significantly higher than the model group and IL-6 was significantly lower than the model group (P<0.05), and the recombinant human osteoprotegerin group IL-4 was significantly higher after the intervention. IL-6 was significantly lower in the tinidazole group than in the tinidazole group (P<0.05). After the intervention, the tinidazole group and the recombinant human osteoprotegerin group were significantly reduced, and the recombinant human osteoprotegerin group RAKNL and MCP-1 were significantly lower than the model group (P>0.05). Conclusion: Recombinant human osteoprotegerin combined with tinidazole has a better therapeutic effect on gums and teeth in mice with periodontitis, and can lower the levels of RAKNL and MCP-1 in serum, inhibit bone resorption and protect teeth.

核因子κB受体活化因子配体抑制剂地舒单抗在肺癌骨转移中的应用

目前,随着基因靶向治疗和免疫治疗在肺癌领域中取得巨大突破,晚期肺癌患者总生存期(OS)显著延长,但发生远处骨转移及骨相关事件(SRE),如骨痛、病理性骨折、脊髓压迫、高钙血症等风险也随之增大,严重影响了患者的生活质量。因此,在全身治疗基础上应积极预防和治疗骨转移及SRE治疗。临床研究表明,核因子κB受体活化因子(RANK)配体(RANKL)/RANK/骨保护素信号通路在肿瘤骨转移中发挥着重要作用,阻断该通路能有效预防和治疗SRE。RANKL抑制剂地舒单抗(商品名:安加维)是一种人免疫球蛋白G2单克隆抗体,可特异性结合RANKL而阻断破骨细胞参与的RANKL/RANK/骨保护素信号通路激活,最终发挥预防骨转移及SRE的作用。与双磷酸盐类药物比较,地舒单抗疗效显著,能明显延长患者OS和SRE的发生时间。同时,地舒单抗还具有抗肿瘤作用。该文就地舒单抗的作用机制、临床研究及安全性等方面的最新研究进行了阐述,以期为临床医生提供参考。

幼鼠脓毒症模型中Rankl调控胸腺自然调节性T细胞免疫平衡的作用机制研究

目的探讨核因子κB受体激活因子配体(receptor activator of nuclear factor kappa-B ligand,Rankl)对脓毒症幼鼠胸腺自然调节性T细胞(regulatory T cell,Treg)的免疫调控机制。方法采用脂多糖(lipopolysaccharide,LPS)腹腔注射构建SD雄性幼鼠脓毒症模型。48只幼鼠按LPS浓度梯度分为4组,分别注射生理盐水(对照组)和5、8、10 mg/kg LPS,每组12只。24只幼鼠按给药分组,随机分为对照组、脓毒症组、野黄芩苷(Rankl抑制剂)组、白桦脂酸(NF-κB激动剂)组共4组,每组6只。取胸腺组织行免疫组化染色,计算Rankl、自身免疫调节因子(autoimmunity regulator,Aire)、叉头翼状螺旋转录因子(forkhead box P3,Foxp3)阳性细胞的平均光密度。胸腺组织匀浆免疫印迹法测定Rankl、核因子κB(nuclear factor kappa-B,NF-κB)p65、Aire、Foxp3的蛋白表达水平。统计学分析采用单因素方差分析和t检验。结果不同浓度梯度模型中,对照组与腹腔注射LPS 5、8、10 mg/kg组,48 h时胸腺组织阳性细胞平均光密度比较[Rankl:(0.209±0.021)%、(0.256±0.008)%、(0.302±0.015)%、(0.361±0.023)%;Aire:(0.279±0.017)%、(0.350±0.021)%、(0.402±0.013)%、(0.459±0.024)%;Foxp3:(0.207±0.014)%、(0.237±0.010)%、(0.277±0.011)%、(0.310±0.016)%],蛋白表达水平比较(Rankl:0.577±0.079、0.740±0.079、0.935±0.043、1.240±0.109;NF-κBp 65:0.150±0.064、0.306±0.055、0.534±0.077、0.949±0.077;Aire:0.324±0.039、0.571±0.062、0.737±0.091、1.019±0.120;Foxp3:0.226±0.098、0.475±0.035、0.824±0.070、1.148±0.087),LPS造模组均高于对照组(P值均<0.05);且随LPS剂量增加而增加,10 mg/kg组增加最明显。不同给药模型中,脓毒症组、野黄芩苷组、白桦脂酸组的胸腺组织阳性细胞平均光密度比较[Rankl:(0.343±0.022)%、(0.262±0.014)%、(0.299±0.020)%;Foxp3:(0.380±0.016)%、(0.340±0.013)%、(0.426±0.012)%;Aire:(0.671±0.079)%、(0.437±0.109)%、(0.893±0.034)%],蛋白表达水平比较(Rankl:0.945±0.059、0.626±0.072、0.801±0.052;NF-κB p65:0.671±0.079、0.633±0.191、1.229±0.106;Aire:0.815±0.144、0.437±0.109、1.219±0.114;Foxp3:0.773±0.093、0.453±0.143、1.256±0.086),野黄芩苷组均低于脓毒症组,白桦脂酸组除了Rankl,其余指标均高于脓毒症组(P值均<0.05)。结论在幼鼠脓毒症模型中,Rankl通过激活NF-κB/Aire/Foxp3信号通路使胸腺Treg大量增殖,发生免疫失调。Rankl抑制剂野黄芩苷可调节此失衡状态,对脓毒症幼鼠产生保护效应。

淫羊藿对激素性股骨头坏死骨组织OPG/RANKL mRNA表达的影响

目的观察糖皮质激素对大鼠股骨头骨组织中骨保护素(OPG)/核因子kappa B受体活化因子配基(RANKL)mRNA表达的影响及淫羊藿的拮抗作用,探讨淫羊藿预防激素性股骨头坏死的作用效果及其作用机制。方法健康SD大鼠48只,雌雄各半,随机分为激素组、淫羊藿组和对照组,每组16只。激素组和淫羊藿组每只给予肌肉注射醋酸泼尼松龙12.5mg/kg,2次/周,淫羊藿组同时给予淫羊藿提取液1 ml/100 g(相当于0.1 g/ml生药)灌胃,而激素组用生理盐水代替淫羊藿灌胃,1次/d;对照组用生理盐水代替激素和淫羊藿以同样方式肌注和灌胃。4周后取左侧股骨头骨组织石蜡包埋,HE染色,鉴定股骨头坏死情况;取右侧股骨头骨组织提取总RNA,采用实时定量聚合酶链反应技术检测OPG和RANKLmRNA表达水平,计算出OPG/RANKL比值。结果激素组、淫羊藿组、对照组大鼠出现股骨头坏死表现比例分别为71.43%(10/14)、26.67%(4/15)、0(0/16);激素组、淫羊藿组、对照组OPG mRNA表达分别为1.13±0.32、1.47±0.42和1.51±0.43;RANKL mRNA表达分别为2.70±1.24、1.82±0.46和1.55±0.42。激素组OPG和OPG/RANKL比值低于淫羊藿和对照组,而RANKL表达高于淫羊藿组和对照组,差异有统计学意义(P<0.05)。淫羊藿组与对照组比较,差异无统计学意义。结论淫羊藿预防激素性股骨头坏死可能与其拮抗皮质激素导致的OPG/RANKL mRNA表达异常有关。

OPG/RANK/RANKL系统与骨质疏松研究最新进展

骨质疏松是严重威胁中老年人健康的骨科常见病,OPG/RANK/RANKL是参与调节骨重建的最重要的分子系统之一,与骨疾病相关的骨质疏松有密切联系,并已成为药物设计的新靶点.因此,对该系统的深入研究将为骨生理、病理机制阐明及骨疾病防治带来积极影响.