CAJ | 학술논문

目的探讨大鼠创伤性股骨头坏死进程中骨软骨G蛋白偶联受体-48(g protein coupled receptor,GPR48)的表达变化,为进一步理解创伤性股骨头坏死的分子机制提供新线索.方法6个月龄 SD 大鼠33只,随机分实验组(24只)和对照组(9只).实验组采用圆韧带离断结合骨膜剥离切除术,对照组采用假手术作为对照.术后1、2和4周时收集股骨头,通过大体、组织学及 Masson 染色,观察股骨头坏死进程;用免疫组织化学、RT-PCR、图像分析等方法检测股骨头坏死过程中 GPR48表达变化.结果(1)大体标本观察发现,与对照组相比,实验组1、2周股骨头关节软骨表面略显粗糙,4周时关节软骨出现不同程度的虫蚀样破损.X线片结果显示,实验组1、2周大鼠股骨头改变较轻微,4周时出现密度减低区,骨小梁排列紊乱.HE 染色结果显示,实验组1、2周部分关节标本软骨表面出现破裂、剥脱;4周时坏死集中在软骨下骨和骺软骨区,但同时可见坏死小梁周围局部成骨细胞活跃迹象,并有少量新生骨形成;3个时间点实验组骨小梁内空骨陷窝率均高于假手术组,并随时间延长呈逐渐升高的趋势,差异有统计学意义(P<0.05).Masson 染色结果显示实验组1、2周时胶原含量下降,骨小梁欠规则;4周骨小梁稀疏、断裂,胶原含量明显减少.(2)免疫组化结果显示,实验组3个时间点 GPR48表达量(IOD 值)逐渐增加,但均低于对照组,差异有统计学意义(P<0.05).(3) RT-PCR 分析结果显示,实验组3个时间点 GPR48 mRNA 表达量逐渐增加,但均低于对照组,差异有统计学意义(P<0.05).结论创伤性股骨头坏死发展进程中,GPR48表达下降,但随坏死进程而逐渐增加,可能与骨坏死后的局部修复有关.
목적탐토대서창상성고골두배사진정중골연골G단백우련수체-48(g protein coupled receptor,GPR48)적표체변화,위진일보리해창상성고골두배사적분자궤제제공신선색.방법6개월령 SD 대서33지,수궤분실험조(24지)화대조조(9지).실험조채용원인대리단결합골막박리절제술,대조조채용가수술작위대조.술후1、2화4주시수집고골두,통과대체、조직학급 Masson 염색,관찰고골두배사진정;용면역조직화학、RT-PCR、도상분석등방법검측고골두배사과정중 GPR48표체변화.결과(1)대체표본관찰발현,여대조조상비,실험조1、2주고골두관절연골표면략현조조,4주시관절연골출현불동정도적충식양파손.X선편결과현시,실험조1、2주대서고골두개변교경미,4주시출현밀도감저구,골소량배렬문란.HE 염색결과현시,실험조1、2주부분관절표본연골표면출현파렬、박탈;4주시배사집중재연골하골화후연골구,단동시가견배사소량주위국부성골세포활약적상,병유소량신생골형성;3개시간점실험조골소량내공골함와솔균고우가수술조,병수시간연장정축점승고적추세,차이유통계학의의(P<0.05).Masson 염색결과현시실험조1、2주시효원함량하강,골소량흠규칙;4주골소량희소、단렬,효원함량명현감소.(2)면역조화결과현시,실험조3개시간점 GPR48표체량(IOD 치)축점증가,단균저우대조조,차이유통계학의의(P<0.05).(3) RT-PCR 분석결과현시,실험조3개시간점 GPR48 mRNA 표체량축점증가,단균저우대조조,차이유통계학의의(P<0.05).결론창상성고골두배사발전진정중,GPR48표체하강,단수배사진정이축점증가,가능여골배사후적국부수복유관.
Objective To explore the variation of the expression of G protein coupled receptor 48 (GPR48) in the progress of traumatic necrosis of the femoral head in rats, which would provide novel clues for further understanding the molecular mechanism of traumatic necrosis of the femoral head. Methods 33 6-month-old Sprague-Dawley (SD) rats were randomly divided into 2 groups. The experimental group (24 rats) were operated through cutting the ligamentum teres and stripping the periosteum, and the control group (9 rats) were performed sham operation. The femoral heads were taken at 3 time points of 1w, 2w and 4w after the operation, and then the samples were analyzed by gross observation, histological analysis and Masson staining to observe the progress of necrosis of the femoral head. Immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PRC), graphical analysis and so on were used to detect the variation of the expression of GPR48 during the process of traumatic necrosis of the femoral head. Results (1) Gross observation showed in the experimental group the cartilage surface of the femoral head was more coarse at 1w and 2w after the operation, and there were more worm-eaten-like breakages at different degrees on the articular cartilage at 4w, when compared with that of the control group. The X-ray in the experimental group showed at 1w and 2w the femoral head in rats was slightly changed, but at 4w the trabecular bone was disordered with irregular organization and some zones of decreased bone mass. The results of HE staining displayed that in the experimental group at 1w and 2w the cartilage surface revealed some small cracks and breakages. At 4w, the necrosis area was centered in the articular cartilage and femoral epiphyseal cartilage, with more active osteoblasts partially around the necrotic trabecular bone and a little new bone formation. For the assay of the rate of empty lacunae, at the 3 time points in the experimental group, all these data were higher than that in the sham group (P<0.05), and with the time going, the rate of empty lacunae was generally increased with statistically significant differences (P<0.05). The results of Masson staining showed that at 1w and 2w the collagen contents in the experimental group was decreased with irregular trabecular bone, but at 4w the trabecular bone became sparse and broken with significant decrease in the collagen content. (2) Immunohistochemical analysis displayed that the GPR48 expression (integrated optical density [IOD] value) in the experimental group at the 3 time points was gradually elevated but significantly lower than that in the sham group, and the differences were statistically significant (P<0.05). (3) RT-PCR analysis revealed that the GPR48 mRNA expression (IOD value) in the experimental group at the 3 time points were also gradually elevated but significantly lower than that in the sham group, and the differences were statistically significant (P<0.05). Conclusions In the progress of traumatic necrosis of the femoral head, the expression of GPR48 is declined, however, it will gradually increase with the development of necrosis, which may be related to the local repair of necrotic bone.