Role of aquaporin-3 in hyperosmolarity-induced apoptosis of rat nucleus pulposus cells <italic style="font-style: italic">in vitro</italic>

Zetong ZHANG; Lang FANG; Yibo GAN; Pei LI; Jixing YE; Chen ZHAO; Qiang ZHOU

doi:10.16016/j.1000-5404.201912129

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Role of aquaporin-3 in hyperosmolarity-induced apoptosis of rat nucleus pulposus cells in vitro

Basic Medicine | 更新时间：2026-01-09

- Role of aquaporin-3 in hyperosmolarity-induced apoptosis of rat nucleus pulposus cells in vitro
- Journal of Third Military Medical University Vol. 42, Issue 9, Pages: 915-922(2020)
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)第一附属医院骨科
  2.401120 重庆，重庆医科大学附属第三医院骨与创伤中心
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81772378);the Basic Research Military-Civilian Integration Project of Southwest Hospital(SWH2017YBXM-20)
- DOI：10.16016/j.1000-5404.201912129
  CLC： R322.72;R329.27;R341
- Received：13 December 2019，
  
  Revised：2020-02-02，
  
  Published：15 May 2020
- 稿件说明：
移动端阅览
Zetong ZHANG, Lang FANG, Yibo GAN, et al. Role of aquaporin-3 in hyperosmolarity-induced apoptosis of rat nucleus pulposus cells in vitro[J]. Journal of Third Military Medical University, 2020, 42(9): 915-922.
DOI：

Zetong ZHANG, Lang FANG, Yibo GAN, et al. Role of aquaporin-3 in hyperosmolarity-induced apoptosis of rat nucleus pulposus cells in vitro[J]. Journal of Third Military Medical University, 2020, 42(9): 915-922. DOI： 10.16016/j.1000-5404.201912129.

摘要

目的

研究水通道蛋白3(aquaporin-3，AQP-3)在高渗透压诱导的大鼠髓核细胞(rat nucleus pulposus)凋亡中的作用。

方法

体外培养的大鼠髓核细胞，按330、550 mOsm/kg的渗透压分组，CCK-8检测髓核细胞增殖，流式细胞分析细胞凋亡率，Western blot和定量PCR检测凋亡相关分子Bcl-2、Bax和caspase3表达情况；使用免疫细胞化学、Western blot和定量PCR检测AQP-3的表达。利用过表达慢病毒提高AQP-3的表达，进一步观察AQP-3对高渗环境诱导凋亡的影响。

结果

相比于330 mOsm/kg渗透压，550 mOsm/kg高渗环境中髓核细胞培养12、24、48、72 h细胞增殖显著降低，髓核细胞凋亡率显著增加(

0.05)，蛋白和mRNA表达结果显示Bcl-2表达明显降低(

0.05)，Bax和cleaved caspase 3/caspase 3表达明显增加(

0.05)。相比于高渗环境中的空白对照髓核细胞，AQP-3过表达髓核细胞的凋亡率显著降低(

0.05)，蛋白和mRNA表达结果显示Bcl-2表达明显增加(

0.05)，Bax和cleaved caspase 3/caspase 3表达明显降低(

0.05)。

结论

高渗透压显著促进髓核细胞的凋亡并降低AQP-3的表达；过表达AQP-3可缓解高渗透压环境下髓核细胞凋亡。

Abstract

Objective

To investigate the role of aquaporin-3 (AQP-3) in hyperosmolarity-induced apoptosis of cultured rat nucleus pulposus (NP) cells.

Methods

Rat NP cells were cultured under different osmotic pressures (330 and 550 mOsm/kg)

and the proliferation and apoptotic rate of the cells were determined using CCK-8 assay and flow cytometry

respectively. Western blotting and real-time quantitative PCR were used to detect the expression levels of apoptosis-related molecules (Bcl-2

Bax

cleaved caspase 3/caspase 3)

and immunocytochemistry

Western blotting and real-time quantitative PCR were used to detect the expression of AQP-3 in the cells. The effect of lentivirus-mediated AQP-3 overexpression on apoptosis of NP cells was investigated after hyperosmolar exposure.

Results

An osmotic pressure of 550 mOsm/kg

as compared with 330 mOsm/kg

significantly decreased NP cell proliferation

increased the

cell apoptotic rate

upregulated the expression of the pro-apoptotic molecules Bax and cleaved caspase 3/caspase 3 (

0.05)

and down-regulated the expression of the anti-apoptotic molecule Bcl-2 (

0.05). The cells with lentivirus-mediated AQP-3 overexpression showed an obviously lowered apoptotic rate following the hyperosmolar exposure and significantly reduced expression of Bax and cleaved caspase 3/caspase 3 (

0.05) with an enhanced expression of Bcl-2 (

0.05).

Conclusion

Hyperosmolar exposure significantly promotes apoptosis of NP cells and reduce the expression of AQP-3. AQP-3 overexpression can partly attenuate apoptosis of NP cells following a hyperosmolar exposure.

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references

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Role of aquaporin-3 in hyperosmolarity-induced apoptosis of rat nucleus pulposus cells in vitro

DOI：10.16016/j.1000-5404.201912129

摘要

Abstract

关键词

Keywords

references