人退变髓核细胞中内质网应激水平的观察研究

文天用; 刘月; 程实; 应金威; 阮狄克

doi:10.16016/j.1000-5404.201803077

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人退变髓核细胞中内质网应激水平的观察研究

临床医学 | 更新时间：2026-01-09

- 人退变髓核细胞中内质网应激水平的观察研究
- Endoplasmic reticulum stress in degenerative human nucleus pulposus cells
- 第三军医大学学报 2018年40卷第12期页码：1098-1103
- 作者机构：
  
  1.710032 西安，空军军医大学西京医院骨科
  2.100048 北京，海军总医院骨科
- 作者简介：
  
  [ 阮狄克，E-mail：ruandikengh@163.com RUAN Dike, E-mail:ruandikengh@163.com
- 基金信息：
  
  国家自然科学基金面上项目(81472121);海军总医院创新培育基金(CXPY201402)
- DOI：10.16016/j.1000-5404.201803077
  中图分类号： R361.3;R394.3;R681.53
- 收稿：2018-03-11，
  
  修回：2018-04-03，
  
  纸质出版：2018-01
- 稿件说明：
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文天用, 刘月, 程实, 等. 人退变髓核细胞中内质网应激水平的观察研究[J]. 第三军医大学学报, 2018,40(12):1098-1103.

Tianyong WEN, Yue LIU, Shi CHENG, et al. Endoplasmic reticulum stress in degenerative human nucleus pulposus cells[J]. Journal of Third Military Medical University, 2018, 40(12): 1098-1103.
文天用, 刘月, 程实, 等. 人退变髓核细胞中内质网应激水平的观察研究[J]. 第三军医大学学报, 2018,40(12):1098-1103. DOI： 10.16016/j.1000-5404.201803077.

Tianyong WEN, Yue LIU, Shi CHENG, et al. Endoplasmic reticulum stress in degenerative human nucleus pulposus cells[J]. Journal of Third Military Medical University, 2018, 40(12): 1098-1103. DOI： 10.16016/j.1000-5404.201803077.

摘要

目的

观察探讨退变与正常髓核组织中内质网应激水平的差异。

方法

获取正常和退变髓核组织共9例(均为海军总医院骨科2016年1月至2017年6月期间住院手术患者)，取组织及分离培养髓核细胞，通过透射电镜观察内质网形态、免疫组化法对髓核组织中葡萄糖调节蛋白78(glucose regulated protein 78

GRP78)、C/EBP同源蛋白(C/EBP homologous protein，CHOP)的表达分布进行定性分析，RT-PCR及Western blot对髓核细胞中GRP78、CHOP进行mRNA和蛋白质水平的分析。

结果

透射电镜结果显示退变髓核细胞的内质网扩张、肿胀，正常片状折叠结构消失；免疫组化染色及Western blot检测结果均可见退变髓核组织GRP78、CHOP的表达升高；RT-PCR提示较之于正常组，退变组GRP78[(1.076±0.13)

(2.471±0.53)

]

、CHOP[(1.09±0.21)

(6.873±1.4)

]

明显升高(

0.05)。

结论

退变髓核细胞中内质网应激水平较正常髓核细胞升高，提示内质网应激可能在髓核细胞退变过程中发挥作用。

Abstract

Objective

To investigate the difference in the levels of endoplasmic reticulum (ER) stress between degenerative and normal human nucleus pulposus cells.

Methods

Nucleus pulposus tissues were collected from specimens of normal and degenerative intervertebral discs from 9 surgical patients (mean age 33 years) admitted in the Department of Orthopedic Surgery

Navy General Hospital between January

2016 and June

2017. The nucleus pulposus cells (NPCs) were isolated and cultured

and the morphology of the ER was observed using transmission electron microscopy (TEM). The expression levels of 2 ER stress-related molecules

namely glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP)

were analyzed using immunohistochemical assay

real-time PCR and Western blotting.

Results

TEM revealed obvious ER dilation and swelling in degenerative NPCs as compared with normal NPCs. Immunohistochemical assay and Western blotting showed obviously increased expressions of GRP78 and CHOP proteins in degenerative NPCs. GRP78 and CHOP mRNA expressions were also significantly increased in degenerative as compared with normal NPCs (2.471±0.53

1.076±0.13

and 6.873±1.4

1.09±0.21

respectively;

0.05).

Conclusion

The abnormal morphology of ER and increased expression of ER stress markers in degenerative NPCs suggest the involvement of ER stress in the development of NPC degeneration.

关键词

Keywords

references

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