CAJ | 학술논문

背景：骨折愈合是成骨细胞与破骨细胞耦联相互作用相互影响促进骨质生长的过程，其中成骨细胞介导骨的形成，破骨细胞介导骨吸收，使骨重建处于一种动态平衡中，于动态平衡环境中促进骨生长，而目前研究多数倾向于单独的成骨或者破骨机制，但会忽略这两种细胞共同存在的环境下相互作用机制。 　　目的：观察补肾接骨中药对成骨与破骨细胞耦联中骨护骨素-核因子κB受体激活剂的配基-核因κB受体活化因子的影响及其在骨折治疗中的作用机制。 　　方法：分离小鼠成骨、破骨细胞并体外细胞培养，建立小鼠“成骨-破骨细胞共育系”作为研究平台，补肾接骨中药1.25，2.5，6.25 g/(kg?d)灌胃，空白对照组给予同等体积的生理盐水灌胃，每日1次，连续7 d。 　　结果与结论：共育培养24 h后，成骨细胞内碱性磷酸酶水平明显高于单纯培养的成骨细胞(P <0.05)。实时PCR检测显示，共育体系细胞中碱性磷酸酶、Runx2及骨护骨素表达增加(P <0.05)，呈剂量依赖性(P <0.05)。Western-blot检测显示，高浓度[6.25 g/(kg?d)]中药能明显促进骨护骨素、核因子κB受体激活剂的配基的表达，抑制核因子κB受体活化因子蛋白表达(P <0.05)。结果证实，补肾接骨中药可动态调节骨护骨素-核因子κB受体激活剂的配基-核因子κB受体活化因子信号通路，对促进骨组织重建与恢复有着积极的作用。
배경：골절유합시성골세포여파골세포우련상호작용상호영향촉진골질생장적과정，기중성골세포개도골적형성，파골세포개도골흡수，사골중건처우일충동태평형중，우동태평형배경중촉진골생장，이목전연구다수경향우단독적성골혹자파골궤제，단회홀략저량충세포공동존재적배경하상호작용궤제。 　　목적：관찰보신접골중약대성골여파골세포우련중골호골소-핵인자κB수체격활제적배기-핵인κB수체활화인자적영향급기재골절치료중적작용궤제。 　　방법：분리소서성골、파골세포병체외세포배양，건립소서“성골-파골세포공육계”작위연구평태，보신접골중약1.25，2.5，6.25 g/(kg?d)관위，공백대조조급여동등체적적생리염수관위，매일1차，련속7 d。 　　결과여결론：공육배양24 h후，성골세포내감성린산매수평명현고우단순배양적성골세포(P <0.05)。실시PCR검측현시，공육체계세포중감성린산매、Runx2급골호골소표체증가(P <0.05)，정제량의뢰성(P <0.05)。Western-blot검측현시，고농도[6.25 g/(kg?d)]중약능명현촉진골호골소、핵인자κB수체격활제적배기적표체，억제핵인자κB수체활화인자단백표체(P <0.05)。결과증실，보신접골중약가동태조절골호골소-핵인자κB수체격활제적배기-핵인자κB수체활화인자신호통로，대촉진골조직중건여회복유착적겁적작용。
BACKGROUND:Fracture healing is the coupling interaction of osteoblasts and osteoclasts that promotes bone growth, in which osteoblast-mediated bone resorption and osteoclasts-mediated bone reconstruction make the bone reconstruction in a dynamic equilibrium to promote bone growth. However, the majorities of the current studies focus on osteogenic or osteoclastic mechanism alone, and ignore the interaction mechanism between these two cels under co-existing conditions. OBJECTIVE:To investigate the effects of Kidney Chinese Herbs on osteoblasts and osteoclasts coupling of osteoprotegerin-receptor activator of nuclear factor kappaB ligand-receptor activator of nuclear factor kappaB and its mechanism of action in fracture treatment. METHODS: Mouse osteoblasts and osteoclasts were isolated and cultured in vitro to establish the mouse “osteoblast-osteoclast co-culture system” as a research platform. Then, Kidney Chinese Herbs at doses of 1.25, 2.5, 6.25 g/(kg?d) were given intragastricaly for 7 consecutive days. Mice in the blank control group were fed with the same volume of normal saline. RESULTS AND CONCLUSION:The alkaline phosphatase activity in osteoblasts co-cultured with osteoclasts was significantly higher than that in osteoblasts cultured alone at 24 hours of culture (P < 0.05). Real-time PCR showed that in the co-culture system, the expression of alkaline phosphatase, Runt related transcription factor 2 and osteoprotegerin were increased in a dose-dependent manner (P < 0.05). Western blot assay showed 6.25 g/(kg?d) Kidney Chinese Herbs could dramaticaly promote the expression of osteoprotegerin and receptor activator of nuclear factor kappaB ligand, but restrained the expression of receptor activator of nuclear factor kappaB (P < 0.05). These findings indicate that Kidney Chinese Herbs can dynamicaly regulate the osteoprotegerin-receptor activator of nuclear factor kappaB ligand-receptor activator signaling pathway, and has a positive effect to promote bone reconstruction and rehabilitation.