Hemeoxygenase-1 protects degenerative human nucleus pulposus cells against apoptosis by NF-κB signaling pathway

Xuemei ZHANG; Wei JIANG

doi:10.16016/j.1000-5404.201711058

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Hemeoxygenase-1 protects degenerative human nucleus pulposus cells against apoptosis by NF-κB signaling pathway

Basic Medicine | 更新时间：2026-01-09

- Hemeoxygenase-1 protects degenerative human nucleus pulposus cells against apoptosis by NF-κB signaling pathway
- Journal of Third Military Medical University Vol. 40, Issue 7, Pages: 561-568(2018)
- 作者机构：
  
  1.400016 重庆，重庆医科大学附属第一医院：产科
  2.400016 重庆，重庆医科大学附属第一医院：骨科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81171751);the Young Scholar of National Natural Science Foundation of China(81501286)
- DOI：10.16016/j.1000-5404.201711058
  CLC： R681.53;R966;R977.3
- Received：08 November 2017，
  
  Revised：2017-12-29，
  
  Published：15 April 2018
- 稿件说明：
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Xuemei ZHANG, Wei JIANG. Hemeoxygenase-1 protects degenerative human nucleus pulposus cells against apoptosis by NF-κB signaling pathway[J]. Journal of Third Military Medical University, 2018, 40(7): 561-568.
DOI：

Xuemei ZHANG, Wei JIANG. Hemeoxygenase-1 protects degenerative human nucleus pulposus cells against apoptosis by NF-κB signaling pathway[J]. Journal of Third Military Medical University, 2018, 40(7): 561-568. DOI： 10.16016/j.1000-5404.201711058.

摘要

目的

探讨血红素氧合酶-1(hemeoxygenase-1，HO-1)对人退变椎间盘髓核细胞凋亡的作用及机制。

方法

采用免疫组化和Western blot检测正常(LVF组，标本来源于年轻的椎体骨折行手术治疗的患者)及退变(IDD组，标本来源于椎间盘退变性疾病行手术治疗的患者)人椎间盘髓核组织中HO-1和磷酸化的P65(p-P65)的表达差异；体外培养退变髓核细胞，通过IL-1β处理增加髓核细胞凋亡，再用载HO-1质粒的慢病毒及小干扰RNA处理髓核细胞后检测凋亡的变化；通过P65抑制剂PDTC抑制p-P65表达后检测髓核细胞凋亡的变化。

结果

免疫组化结果显示，HO-1在IDD组中的阳性表达率为(18.58±0.59)%，明显低于LVF组(58.26±2.48)%(

0.05)，而p-P65在IDD组中的阳性表达率为(40.27±2.03)%，明显高于LVF组(19.75±1.98)%(

0.05)。IL-1β能增加退变髓核细胞的凋亡，同时引起p-P65表达增加，HO-1-siRNA抑制HO-1表达后可以进一步提高髓核细胞凋亡率(

0.05)，而过表达HO-1处理细胞后能够降低IL-1β引起的凋亡增加，同时降低p-P65表达(

0.05)。使用P65抑制剂PDTC抑制髓核细胞P65信号通路后，髓核细胞凋亡率明显降低，此时沉默HO-1不能增加髓核细胞凋亡。

结论

HO-1能够通过NF-κB通路抑制人退变椎间盘髓核细胞凋亡。

Abstract

Objective

To determine the effect of hemeoxygenase-1 (HO-1) on apoptosis in human degenerative nucleus pulposus cells (NPCs)

and investigate the underlying mechanism.

Methods

The expression of HO-1 and nuclear factor kappa B (NF-κB) subunit phosphorylated P65 (p-P65) was determined with immunohistochemical assay and Western blot analysis in human nucleus pulposus tissues from healthy young individuals (due to lumbar vertebral fracture

LVF group) and old patients with intervertebral disc degeneration (IDD group). After the degenerative NPCs were cultured

in vitro

and IL-1β was added to induce cell apoptosis measured by annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) double staining analysis. The effects of lentiviral vector of HO-1 and its siRNA on the cell apoptosis were also determined. The apoptosis was also observed after the treatment of p65 inhibitor PDTC.

Results

The positive rate of HO-1 was (18.58±0.59)% in the IDD

group

significantly lower than that in the LVF group [(58.26±2.48)%

0.05

]

and opposite phenomenon was found in that of p-P65 [(40.27±2.03)%

(19.75±1.98)%

0.05

]

. IL-1β enhanced the cell apoptosis and up-regulated the expression of p-P65 in the degenerative NPCs. HO-1-siRNA inhibited the HO-1 expression and then promoted cell apoptosis (

0.05). Overexpression of HO-1 by LV-HO-1 inhibited the increased NPCs apoptosis after IL-1β treatment

and simultaneously inhibited the expression of p-P65 in human NPCs (

0.05). Furthermore

the NF-κB inhibitor

PDTC

was used to treat NPCs

the number of apoptotic NPCs was significantly decreased no matter with or without HO-1-siRNA transfection.

Conclusion

HO-1 inhibits apoptosis in degenerative human NPCs through NF-κB pathway.

关键词

Keywords

references

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