青岛大学医学院学报2017，Vol.53Issue(3)：257-260,4.DOI:10.13361/j.qdyxy.201703002
椎间盘髓核细胞应力模型中Piezo1蛋白的表达
EXPRESSION OF PIEZO1 PROTEIN IN STRESS MODEL OF NUCLEUS PULPOSUS CELLS OF THE INTERVERTEBRAL DISC
王天宝 1李晓飞 1冷萍 2马佳超 1张海宁1
作者信息
- 1. 青岛大学附属医院关节外科,山东青岛266003
- 2. 青岛大学附属医院药剂科,山东青岛266003
- 折叠
摘要
Abstract
Objective To investigate the expression of a new mechanically activated ion channel,Piezo1 protein,in a stress model of human nucleus pulposus cells of the intervertebral disc.Methods The human nucleus pulposus cells of the intervertebral disc were divided into stretch stress group and stretch stress+ Piezo1 protein inhibitor group.An in vitro culture-stress stimulation model of human nucleus pulposus cells of the intervertebral disc was established,and FX-4000T was used for the treatment of nucleus pulposus cells.Both groups were treated with 0-,2-,12-,24-,and 48 h cyclic stretch stress.RT-PCR and Western blot were used to measure the mRNA and protein expression of Piezo1,and a laser scanning confocal microscope was used to measure the fluorescence intensity of Piezo1 protein.Results The results of RT-PCR showed that the stretch stress group with 2-hour cyclic stretch stress had a significant increase in the mRNA expression of Piezo1 compared with the group with 0-hour cyclic stretch stress;the stretch stress group with 24-hour cyclic stretch stress had higher mRNA expression of Piezo1 than the other groups,and compared with this group,the stretch stress group with 48-hour cyclic stretch stress had a significant reduction in the mRNA expression of Piezo 1 (F=13.917,P＜0.05).The results of Western blot showed that the stretch stress group with 2-hour cyclic stretch stress had a slight increase in the protein expression of Piezo1 compared with the group with 0-hour cyclic stretch stress;the stretch stress group with 24-hour cyclic stretch stress had higher protein expression of Piezo1 than the other groups,and compared with this group,the stretch stress group with 48 h cyclic stretch stress had a significant reduction in the protein expression of Piezo1 (F=12.781,P＜0.05).The stretch stress+Piezo1 protein inhibitor group had significantly lower mRNA and protein expression of Piezo1 than the stretch stress group.Piezo1 protein was expressed in the cytoplasm and nucleus of the nucleus pulposus cells and had strong fluorescence.Conclusion Piezo1 protein is expressed stably in human nucleus pulposus cells and its expression is associated with stress loading time.关键词
机械激活性离子通道/椎间盘移位/椎间盘髓核细胞Key words
mechanically sensitive ion channel/intervertebral disc displacement/intervertebral disc nucleus pulposus cells分类
医药卫生引用本文复制引用
王天宝,李晓飞,冷萍,马佳超,张海宁..椎间盘髓核细胞应力模型中Piezo1蛋白的表达[J].青岛大学医学院学报,2017,53(3):257-260,4.
