[1]张泽桐,房朗,甘翼博,等.水通道蛋白3在高渗环境诱导髓核细胞凋亡中的作用[J].第三军医大学学报,2020,42(09):915-922.
 ZHANG Zetong,FANG Lang,GAN Yibo,et al.Role of aquaporin-3 in hyperosmolarity-induced apoptosis of rat nucleus pulposus cells in vitro[J].J Third Mil Med Univ,2020,42(09):915-922.
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水通道蛋白3在高渗环境诱导髓核细胞凋亡中的作用(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第09期
页码:
915-922
栏目:
基础医学
出版日期:
2020-05-15

文章信息/Info

Title:
Role of aquaporin-3 in hyperosmolarity-induced apoptosis of rat nucleus pulposus cells in vitro
作者:
张泽桐房朗甘翼博李培叶吉星赵晨周强
陆军军医大学(第三军医大学)第一附属医院骨科;重庆医科大学附属第三医院骨与创伤中心
Author(s):
ZHANG Zetong FANG Lang GAN Yibo LI Pei YE Jixing ZHAO Chen ZHOU Qiang

Department of Orthopedics, First Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400038; Department of Orthopedics, the Third Affiliated Hospital of Chongqing Medical University, Chongqing, 401120, China

关键词:
水通道蛋白渗透压凋亡髓核细胞椎间盘退变
Keywords:
aquaporin osmolarity apoptosis nucleus pulposus intervertebral disc degeneration  
分类号:
R322.72; R329.27; R341
文献标志码:
A
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(P<0.05),蛋白和mRNA表达结果显示Bcl-2表达明显降低(P<0.05),Bax和cleaved caspase 3/caspase 3表达明显增加(P<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(P<0.05),蛋白和mRNA表达结果显示Bcl-2表达明显增加(P<0.05),Bax和cleaved caspase 3/caspase 3表达明显降低(P<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:

ObjectiveTo investigate the role of aquaporin-3 (AQP-3) in hyperosmolarity-induced apoptosis of cultured rat nucleus pulposus (NP) cells. MethodsRat 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. ResultsAn 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 (P<0.05), and down-regulated the expression of the anti-apoptotic molecule Bcl-2 (P<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 (P<0.05) with an enhanced expression of Bcl-2 (P<0.05). ConclusionHyperosmolar 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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更新日期/Last Update: 2020-05-06