摘要
目的研究CM-272对破骨细胞活化及骨质疏松性骨丢失的作用。方法(1)体外实验:小鼠骨髓腔提取BMMs细胞,使用不同剂量CM-272干预BMMs细胞,CCK-8检测确定安全剂量。诱导BMMs细胞破骨分化,使用高低剂量CM-272干预诱导过程,3d时检测氧化应激状态,5d进行TRAP染色。(2)体内实验:随机将28只C57BL/6J小鼠分为4组,假手术组、骨松组、低剂量组、高剂量组,造模休养1周后,每周尾静脉分别注射生理盐水、1mg/kg及2.5mg/kgCM-272,空白组于相同时间尾静脉注射生理盐水。给药8周后,使用micro-CT进行扫描,并进行骨参数分析。结果(1)体外实验:低于0.5μmol/L剂量的CM-272对BMMs无明显抑制作用,使用0.5及0.1μmol/L作为研究的高低剂量;CM-272可抑制RANKL诱导的破骨细胞活化,并且这种抑制作用呈现剂量依赖性;CM-272可以抑制破骨细胞活化过程中的ROS蓄积。(2)体内实验:去势导致小鼠骨质疏松性骨丢失,高剂量CM-272明显缓解此过程,提升BMD、BV/TV、TB.N,改善骨微结构。结论CM-272通过降低胞内ROS异常蓄积,缓解破骨细胞的活化,以及由此导致的骨质疏松性骨丢失。
Objective To study the effect of CM-272 on osteoclast activation and osteoporotic bone loss.Methods(1)In vitro experiments:BMMs cells were extracted from the bone marrow cavity of mice,and different doses of CM-272 were used to interfere BMMs cells,and CCK-8 assay was performed to determine the safe dose.BMMs cells were induced to osteoclast differentiation using high and low doses of CM-272 in the induction process,and oxidative stress status was detected at 3 days and TRAP staining was performed at 5 days.(2)In vivo experiments:28 C57BL/6J mice were randomly divided into 4 groups,control group,osteoporosis group,lowdose group and high-dose group.1 week after ovariectomy,saline,1 mg/kg and 2.5 mg/kg CM272 were injected into the tail vein of the osteoporosis group,low-dose group and high-dose group mice weekly respectively,and saline was injected into the tail vein of the control group at the same time.After 8 weeks of administration,scans were performed using micro-CT and bone parameters were analyzed.Results(1)In vitro experiments:CM-272 at doses below 0.5μmol/L had no significant inhibitory effect on BMMs,and 0.5 and 0.1μmol/L were used as the high and low doses for the study.CM-272 inhibited RANKL-induced osteoclast activation,and the inhibition showed a dose-dependent effect.CM-272 could inhibit ROS accumulation during osteoclast activation.(2)In vivo experiments:ovariectomy leads to osteoporotic bone loss in mice,and high doses of CM-272 significantly alleviates this process,elevate BMD,BV/TV,TB.N,and improves bone microarchitecture.Conclusion The results suggest that CM-272 alleviates osteoclast activation and the subsequent osteoporotic bone loss by reducing abnormal intracellular ROS accumulation.
作者
张巍
杨鹏
俞磊
李文明
耿德春
ZHANG Wei;YANG Peng;YU Lei;LI Wen-ming;GENG De-chun(Department of Orthopedics,First Affiliated Hospital of Soochow University,Suzhou,Jiangsu,215006,China)
出处
《中国血液流变学杂志》
CAS
2023年第1期1-5,F0002,共6页
Chinese Journal of Hemorheology
基金
国家自然科学基金资助项目(82072425,82072498,82272157)。
