[1]王天宝,李晓飞,冷萍,等.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(03):257-260.[doi:10.13361/j.qdyxy.201703002]
 WANG Tianbao,LI Xiaofei,LENG Ping,et al.EXPRESSION OF PIEZO1 PROTEIN IN STRESS MODEL OF NUCLEUS PULPOSUS CELLS OF THE INTERVERTEBRAL DISC[J].Acta Aacademiae Medicinae Qingdao,2017,53(03):257-260.[doi:10.13361/j.qdyxy.201703002]
点击复制

椎间盘髓核细胞应力模型中Piezo1蛋白的表达()
分享到:

《青岛大学医学院学报》[ISSN:1672-4488/CN:37-1356/R]

卷:
第53卷
期数:
2017年03期
页码:
257-260
栏目:
出版日期:
2017-08-07

文章信息/Info

Title:
EXPRESSION OF PIEZO1 PROTEIN IN STRESS MODEL OF NUCLEUS PULPOSUS CELLS OF THE INTERVERTEBRAL DISC
文章编号:
1672-4488(2017)03-0257-04
作者:
王天宝1李晓飞1冷萍2马佳超1张海宁1
青岛大学附属医院,山东 青岛 266003 1 关节外科; 2 药剂科
Author(s):
WANG Tianbao LI Xiaofei LENG Ping MA Jiachao ZHANG Haining
Department of Joint Surgery, The Afflicated Hospital of Qingdao University, Qingdao 266003, China
关键词:
机械激活性离子通道椎间盘移位椎间盘髓核细胞
Keywords:
mechanically sensitive ion channel intervertebral disc displacement intervertebral disc nucleus pulposus cells
分类号:
R329.29
DOI:
10.13361/j.qdyxy.201703002
文献标志码:
A
摘要:
目的 探讨新型机械激活性离子通道Piezo1蛋白在人椎间盘髓核细胞应力模型中的表达。
方法 将人椎间盘髓核细胞分成牵张应力组和牵张应力+Piezo1蛋白抑制剂组,构建人椎间盘髓核细胞体外培养-应力刺激模型,采用多通道细胞牵张应力加载系统FX-4000T处理髓核细胞,两组均加载0、2、12、24、48 h的周期性牵张应力。采用RT-PCR、Western-blot技术检测两组Piezo1的表达水平,应用激光共聚焦显微镜检测Piezo1蛋白荧光的强弱。
结果 RT-PCR结果显示,牵张应力组给予2 h周期性牵张应力时,Piezo1 mRNA表达较给予0 h周期性牵张应力时增加,给予24 h周期性牵张应力时Piezo1 mRNA表达高于其他牵张应力作用时,而给予48 h周期性牵张应力时较给予24 h周期性牵张应力时降低,差异有显著性(F=13.917,P<0.05)。Western-blot结果显示,牵张应力组给予2 h周期性牵张应力时Piezo1蛋白表达较给予0 h周期性牵张应力时略有增加,给予24 h周期性牵张应力时Piezo1蛋白表达高于其他各组,而给予48 h周期性牵张应力时Piezo1蛋白表达较给予24 h周期性牵张应力时降低,差异有显著性(F=12.781,P<0.05)。牵张应力+Piezo1蛋白抑制剂组Piezo1 mRNA及其蛋白表达均明显小于牵张应力组。Piezo1蛋白表达于髓核细胞的细胞质与细胞核,荧光较强。
结论 人椎间盘髓核细胞中Piezo1蛋白稳定表达,并且与应力加载时间相关。
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 Piezo1 (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.
更新日期/Last Update: 2017-08-13