激素性股骨头坏死患者骨髓基质细胞骨保护素/核因子kappa B受体活化因子配基蛋白表达的研究

目的观察激素性股骨头坏死患者骨髓基质细胞（bone marrow stroma cells,BMSCs）骨保护素（osteoprotegerin,OPG）/核因子kappa B受体活化因子配基（receptor activator of nuclear factor kappa B ligand,RANKL）蛋白表达情况,探讨长期应用激素导致股骨头坏死的另一病理机制。方法 2007年3月至2008年3月,取激素性股骨头坏死患者骨髓及股骨头骨组织35例（实验组）,股骨颈骨折患者骨髓及股骨头骨组织21例（对照组）。两组男女比例均为4：3;年龄41～70岁,实验组平均55.34岁,对照组平均55.33岁;实验组最近2年内接受过皮质激素治疗超过3周或超过1周的大剂量冲击治疗,对照组从未接受过超过1周的激素治疗。骨组织标本行多聚甲醛固定后石蜡包埋,HE染色。所取骨髓采用贴壁法分离培养骨髓基质细胞,采用蛋白免疫印迹（Western blot）技术检测骨髓基质细胞OPG和RANKL蛋白表达水平,并得出OPG/RANKL比值。结果 HE染色：实验组未见完整的骨小梁和骨单位,可见不连续的骨碎片,碎片骨陷窝内骨细胞大部分消失,周围大量炎性肉芽组织。对照组可见完整骨单位由板层骨构成,板层骨连续完整,围绕血管呈同心圆排列,小梁骨陷窝内可见骨细胞。骨髓基质细胞Western blot检测：实验组和对照组OPG/RANKL蛋白表达比分别为1.13±0.65和2.54±0.35,实验组明显低于对照组,有统计学差异（P〈0.05）。结论长期应用糖皮质激素致股骨头坏死可能与其调控骨髓基质细胞OPG和RANKL表达有关。

OPG/RANKL protein expresssion in bone marrow stroma cells of glucocorticoid-induced nerrosis of femoral head patients

Objective To investigate the expression levels of osteoprotegerin（OPG） and receptor activator of nuclear factor kappa B ligand（RANKL） protein in marrow stroma cells of patients suffering giucocorticoid-induced necrosis of femoral head（GNFH）,and to discuss the relationship between OPG/RANKL system and GNFH.Methods Bone tissues and marrow stroma cells of femoral head were collected in 35 patients with GNFH（experiment group） and 21 patients with femoral neck fracture（control group）.The ratio of men to women was 4：3 in the two groups,and the ages of the patients ranged from 41 to 70 years old（average 55.34 years in the experimental group and 55.33 years in the control group）.The experimental group had received over three week＇s glucocorticoid treatment or more than one week＇s high-dose glucocorticoid therapy in recent two years,but the control group had never received more than one week＇s hormone therapy.In two groups,microstructure of bone tissue of femoral head was detected by HE stain.Bone marrow stromal cells（BMSCs） were isolated and cultured by adherent-wall method.the protein expression levels of OPG and RANKL were examined by western blot for each example.Results HE stain：bone trabeculae and bone units were replaced by interrupted bone fragments, which were surrounded by inflammation and granulation tissue.Few osteocyte were seen in bone lacunae in experiment group.In control group,bone trabeculae and bone units were made by complete lamellar bone which surrounded blood vessels and osteocytes were seen in lacunae.Western blot testing;the ratio of OPG/RANKL protein expression was 1.13±0.65 and 2.54±0.35 in experimental group and control group.The ratio of in experiment group was significantly lower than control group（P 0.05 ）.Conclusion The glucocorticoid-induced necrosis of femoral head may be related to the expression levels of OPG/RANKL in marrow stroma cells.