护骨素,κB-受体激活因子及κB-受体激活因子配体在缺血性股骨头坏死组织中的表达特征研究

Expression features of osteoprotegerin,receptor activator of nuclear factor kappa-B and receptor activator of nuclear factor kappa-B ligand in avascular necrosis femoral head

目的探讨护骨素、κB-受体激活因子及κB-受体激活因子配体对缺血性股骨头坏死的保护作用。方法采集缺血性股骨头坏死患者坏死组织和对应的正常组织,提取总RNA及总蛋白样品。q PCR检测坏死组织及正常组织中护骨素（osteoprotegerin,OPG）、κB-受体激活因子（receptor activator of nuclear factor kappa-B,RANK）及κB-受体激活因子配体（receptor activator of nuclear factor kappa-B ligand,RANKL）mRNA表达水平,Western blot检测坏死组织及正常组织中OPG、RANK和RANKL蛋白表达水平。结果q PCR检测结果表明,坏死组织中OPG表达量（4.56±0.37）显著高于正常组织中OPG表达量（3.39±0.52,P〈0.05）。坏死组织和正常组织中RANKL mRNA表达量分别为（0.86±0.11）,（0.31±0.08）,比较差异具有统计学意义（P〈0.05）。坏死组织和正常组织中RANK mRNA表达量分别为（0.87±0.12）,（0.56±0.13）比较差异有统计学意义（P〈0.05）。正常组织中RANKL/OPG和RANK/OPG比值分别0.69,0.52。坏死组织中,RANKL/OPG和RANK/OPG比值分别1.35,0.61。Western blot检测结果表明,坏死组织中OPG的表达量和正常组织中的表达量基本一致。所有的样品中均检测到了RANKL蛋白表达且坏死组织和正常组织中RANKL蛋白表达量基本一样。所有的样品均未检测到RANK蛋白表达。结论 OPG、RANK和RANKL在坏死区域骨重建和骨稳态紊乱过程中发挥重要作用,它们可能对骨损伤及随后的髋关节脱位有影响。

Objective To determine the protective effect of osteoprotegerin（OPG）,receptor activator of nuclear factor kappa-B（RANK） and RANK ligand（RANKL） on avascular necrosis femoral head.Methods Necrotic tissue or corresponding normal tissues were collected from 29 avascular necrosis femoral head patients.Quantitative Real Time PCR（q PCR） is used to evaluate mRNA expression of OPG,RANK and RANKL.OPG and RANKL protein levels were estimated by Western blot.Results The results of q PCR showed that the expression of OPG in the necrotic tissue was significantly higher than that in the normal tissue（4.56±0.37）（3.39±0.52）（P〈0.05）.The expression levels of RANKL mRNA in necrotic tissues and normal tissues were（0.86±0.11） and（0.31±0.08）,respectively,the difference was statistically significant（P〈0.05）.The expression levels of RANK mRNA in necrotic tissues and normal tissmes were（0.87±0.12）,（0.56±0.13） respectively,the difference was statistically significant（P〈0.05）.The ratio of RANKL/OPG and RANK/OPG in normal tissues were 0.69 and 0.52,respectively.In the necrotic tissues,RANKL/OPG and RANK/OPG ratios were 1.35 and 0.61,respectively.Results of Western blot showed that the expression of OPG in necrotic tissues was consistent with that in normal tissues.The expression of RANKL protein was detected in all samples,and the expression of RANKL protein in necrotic tissue and normal tissue was almost the same.RANK protein expression was not detected in all samples.Conclusion OPG,RANK and RANKL play important roles in progress of bone remodeling in necrotic area and in disturbance of bone homeostasis and might have an effect on bone destruction and subsequent collapse of hip joint.