补肾活血方对应力条件下终板软骨细胞生物节律分子BMAL1与CLOCK表达的影响
Effect of Bushen Huoxue Fang(BSHXF)on the expression of circadian rhythm molecules BMAL1 and CLOCK in endplate chondrocytes under mechanical loading
赵柯 1熊勇 2周帅琪 1谢榕 1杜雨轩 1王戈文 1梁煜鹏 1金哲峰 1朱立国 1展嘉文1
作者信息
- 1. 中国中医科学院望京医院,北京 100102
- 2. 湖北中医药大学针灸骨伤学院,湖北武汉 430061
- 折叠
摘要
目的 观察补肾活血方对应力退变终板软骨细胞(CEPCs)生物节律关键分子的调控作用.方法 采用地塞米松同步化处理,检测 CEPCs 48 h内节律分子表达变化;探索不同应力条件(5％或10％牵张力)对CEPCs节律分子表达的影响.筛选异常应力条件,设置空白对照组、异常应力组、补肾活血方干预组、补肾活血方对照组,采用qRT-PCR检测CEPCs节律分子,脑和肌肉芳香烃受体核转运样蛋白1基因(BMAL1)和哺乳动物时钟节律调节因子(CLOCK),以及合成代谢标志物蛋白聚糖(Aggrecan)表达;通过流式细胞术检测各组细胞凋亡情况.结果 CEPCs呈现出周期性节律性变化.生理负荷可维持CEPCs生物节律相位、周期和振幅量.异常应力(10％牵张力、1 Hz,24h)导致BMAL1和CLOCK的表达降低(P＜0.01),伴随Aggrecan表达降低(P＜0.01)与细胞凋亡率增加(P＜0.01).补肾活血方含药血清干预后BMAL1、CLOCK、Aggrecan表达显著增加(P＜0.05),CEPCs凋亡显著降低(P＜0.01).结论 补肾活血方可延缓CEPCs应力退变,并调控生物节律分子BMAL1和CLOCK表达.
Abstract
Objective To observe the role of BSHXF in regulating critical molecules in the circadian rhythms of mechanical load-degenerated endplate chondrocytes(CEPCs).Methods Dexamethasone synchronization treatment was used to detect the changes in the circadian rhythms of CEPCs over 48 h;different mechanical load conditions(5％or 10％draft force)were applied to explore the effects of mechanical load on the expression of rhythmic molecules in CEPCs.Control group,abnormal mechanical load group,intervention group of BHHXF and BHHXF control group were set up.The qRT-PCR was used to detect the expression of crucial rhythm molecules of CEPCs,brain and muscle Arnt-like 1(BMAL1)and circadian locomotor output cycles kaput(CLOCK),as well as the anabolic marker Aggrecan;apoptosis of cells in each group was detected by flow cytometry.Results CEPCs showed periodic rhythmic changes.Physiological loading caused elevated quantification of the phase,period,and amplitude of CEPCs'circadian rhythms.Abnormal mechanical loading(10％draft force,1 Hz,24 h)resulted in decreased expression of BMAL1 and CLOCK(P＜0.01),reduced expression of the Aggrecan(P＜0.01),and increased apoptosis(P＜0.05).The drug-containing serum of BSHXF elevated the expression of BMAL1,CLOCK,and Aggrecan(P＜0.05)and decreased apoptosis of CEPCs(P＜0.01)after mechanical loading regression.Conclusion BSHXF can effectively delay the mechanical load degeneration of CEPCs,and its mechanism of action is related to the regulation of BMAL1 and CLOCK.
关键词
补肾活血方/终板软骨细胞/生物节律/应力退变/应力Key words
Bushen Huoxue Fang/Endplate chondrocytes/Circadian rhythm/Mechanical load degeneration/Mechanical load引用本文复制引用
出版年
2024
NETL
NSTL
万方数据