山羊下颌骨颏孔区成骨细胞增殖节律的实验研究

An Experimental Study on Circadian Rhythm of the Proliferative Index of Mandibular Osteoblast in Goats

目的 研究正常生理条件下山羊下颌骨成骨细胞的增殖指数是否具有昼夜节律性变化的特点。方法 选取 2 4只健康山羊 ,随机分为 6组 ,饲养 1周后分别在 0∶0 0、4∶0 0、8∶0 0、12∶0 0、16∶0 0、2 0∶0 0点处死动物 ,并取右下颌骨颏孔区 2cm× 2cm× 0 5cm骨组织进行原代细胞培养 ,培养成骨细胞 ,采用抗 5_溴脱氧尿嘧啶核苷 (Brdu)单克隆抗体技术结合流式细胞术检测各时间点成骨细胞的增殖指数 (PI) ,并采用单余弦分析法对其进行昼夜节律性分析。结果 山羊下颌骨成骨细胞PI表现为明确的昼低夜高的 2 4h节律变化 (P <0 0 1) ,其峰值出现在 19∶4 4左右 ,谷值出现在晨 7∶4 4左右。结论 正常条件下山羊下颌骨成骨细胞的增殖活性具有昼夜节律性变化的特点 。

Objective\ The aim of this study was to investigate the circadian rhythm of the proliferative index of mandibular osteoblasts of goats in a normal situation.Methods\ Twenty four healthy goats were selected and randomly divided into six groups. They were killed, and a piece of bone tissue with 2 cm×2 cm×0.5 cm was obtained from right mandibular mental foramen region at 0∶00, 4∶00, 8∶00, 12∶00, 16∶00 and 20∶00 time points, after they had been fed for one week. The primary cell culture method was used to cultivate osteoblasts from the bone. The method of anti_5_bromodeoxyuridine monoclonal antibody and the flow cytometric analysis were used to detect the proliferative index of the mandibular osteoblasts at different time. The data were analyzed using the method of single cosine rhythmometry.Results\ The proliferative indexes of the mandibular osteoblasts of goats were higher at the nighttime and lower in the daytime. The undulation of the proliferative indexes displayed a significant circadian rhythm (P<0.01). The peak value was obtained at about 19∶44, and the lowest value at about 7∶44.Conclusion\ The proliferative indexes of the mandibular osteoblasts of goats are not stable within 24 hours in a normal situation, but with significant circadian rhythm. There are different proliferative indexes at different time points.\;