氧化苦参碱通过COX-2/PINK1/Parkin信号通路介导的线粒体自噬对MG63骨肉瘤细胞的促凋亡机制

姬健钧; 邱文奎

doi:10.6039/j.issn.1001-0408.2024.01.08

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氧化苦参碱通过COX-2/PINK1/Parkin信号通路介导的线粒体自噬对MG63骨肉瘤细胞的促凋亡机制

药学研究 | 更新时间：2024-01-11

- 氧化苦参碱通过COX-2/PINK1/Parkin信号通路介导的线粒体自噬对MG63骨肉瘤细胞的促凋亡机制
- Mechanism of oxymatrine promoting the apoptosis of osteosarcoma MG63 cell line through mitophagy mediated by COX-2/PINK1/Parkin signaling pathway
- 中国药房 2024年35卷第1期页码：44-50
- 作者机构：
  
  开封市中心医院骨科二病区，河南开封　475000
- 作者简介：
  
  主治医师，硕士。研究方向：骨科学。E-mail：jijianjun653@163.com
  副主任医师，硕士。研究方向：骨科学。E-mail：526087266@qq.com
- 基金信息：
  
  河南省医学科技攻关计划项目(LHGJ20210834)
- DOI：10.6039/j.issn.1001-0408.2024.01.08
  中图分类号： R965
- 纸质出版日期：2024-01-15，
  
  收稿日期：2023-07-14，
  
  修回日期：2023-11-17，
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姬健钧,邱文奎.氧化苦参碱通过COX-2/PINK1/Parkin信号通路介导的线粒体自噬对MG63骨肉瘤细胞的促凋亡机制 ^Δ[J].中国药房,2024,35(01):44-50.

JI Jianjun,QIU Wenkui.Mechanism of oxymatrine promoting the apoptosis of osteosarcoma MG63 cell line through mitophagy mediated by COX-2/PINK1/Parkin signaling pathway[J].ZHONGGUO YAOFANG,2024,35(01):44-50.
姬健钧,邱文奎.氧化苦参碱通过COX-2/PINK1/Parkin信号通路介导的线粒体自噬对MG63骨肉瘤细胞的促凋亡机制 ^Δ[J].中国药房,2024,35(01):44-50. DOI： 10.6039/j.issn.1001-0408.2024.01.08.

JI Jianjun,QIU Wenkui.Mechanism of oxymatrine promoting the apoptosis of osteosarcoma MG63 cell line through mitophagy mediated by COX-2/PINK1/Parkin signaling pathway[J].ZHONGGUO YAOFANG,2024,35(01):44-50. DOI： 10.6039/j.issn.1001-0408.2024.01.08.

摘要

目的

基于环氧合酶2（COX-2）/PTEN诱导激酶1（PINK1）/帕金森病蛋白2（Parkin）信号通路介导的线粒体自噬研究氧化苦参碱对人骨肉瘤MG63细胞的促凋亡机制。

方法

取MG63细胞经2.0、4.0、8.0 mg/mL的氧化苦参碱和6 μmol/L的5-氟尿嘧啶（5-FU）作用后，检测细胞凋亡率、凋亡相关蛋白[B细胞淋巴瘤2（Bcl-2）、Bcl-2相关X蛋白（Bax）]表达水平、线粒体膜电位降低比例、线粒体自噬水平以及PINK1、Parkin、微管相关蛋白1轻链3Ⅱ（LC3-Ⅱ）蛋白表达水平。采用PINK1小干扰RNA（PINK1 siRNA）干扰PINK1的表达，将细胞分为对照组、PINK1 siRNA组、氧化苦参碱组、PINK1 siRNA+氧化苦参碱组，检测细胞中PINK1、Parkin、LC3-Ⅱ蛋白表达水平和线粒体膜电位降低比例以及细胞凋亡率。采用慢病毒感染使COX-2过表达，将细胞分为对照组、氧化苦参碱组、COX-2组、COX-2+氧化苦参碱组，检测细胞中COX-2、PINK1和Parkin蛋白表达水平以及线粒体膜电位降低比例。

结果

经氧化苦参碱干预后，细胞凋亡率，Bax、PINK1、Parkin、LC3-Ⅱ蛋白表达水平，线粒体自噬水平和线粒体膜电位降低比例均显著升高（

＜0.05），Bcl-2蛋白表达水平显著降低（

＜0.05）。与氧化苦参碱组比较，PINK1 siRNA+氧化苦参碱组细胞中PINK1、Parkin、LC3-Ⅱ蛋白表达水平和细胞凋亡率以及线粒体膜电位降低比例均显著降低（

＜0.05）。与氧化苦参碱组比较，COX-2+氧化苦参碱组细胞中COX-2蛋白表达水平显著升高（

＜0.05），PINK1、Parkin蛋白表达水平和线粒体膜电位降低比例均显著降低（

＜0.05）。

结论

氧化苦参碱可通过抑制COX-2表达，介导线粒体自噬信号通路PINK1/Parkin的过度活化，从而促进骨肉瘤细胞凋亡。

Abstract

OBJECTIVE

To study the mechanism of oxymatrine inducing apoptosis of osteosarcoma MG63 cell line based on mitophagy mediated by cyclooxygenase-2 （COX-2）/PTEN-induced putative kinase-1 （PINK1）/Parkinson disease protein-2 （Parkin） signaling pathway.

METHODS

MG63 cells were treated with 2.0， 4.0， 8.0 mg/mL oxymatrine and 6 μmol/L 5-fluorouracil， then the apoptotic rate， the expression of apoptosis-related proteins [B-cell lymphoma-2 （Bcl-2）， Bcl-2 related X protein （Bax）]， the proportion of decrease in mitochondrial membrane potential， the level of mitophagy as well as the protein expressions of PINK1， Parkin， and microtubule-associated protein 1 light chain-3Ⅱ （LC3-Ⅱ） were detected. PINK1 small interfering RNA （siRNA） was adopted to intervene in the expression of PINK1， the cells were divided into control group， PINK1 siRNA group， oxymatrine group， and PINK1 siRNA+oxymatrine group； the protein expressions of PINK1， Parkin， and LC3-Ⅱ， the proportion of decrease in mitochondrial membrane potential （MMP） as well as apoptotic rate were detected. The lentivirus infection technique was used to overexpress COX-2， the cells were divided into control group， oxymatrine group， COX-2 group， and COX-2+oxymatrine group. The protein expressions of COX-2， PINK1， and Parkin， as well as the proportion of decrease in MMP were detected.

RESULTS

After being treated with oxymatrine， the apoptotic rate， the protein expressions of Bax， PINK1， Parkin， and LC3-Ⅱ， the level of mitophagy as well as the proportion of decrease in MMP were significantly increased （

＜0.05）， while the protein expression of Bcl-2 was significantly decreased （

＜0.05）. Compared with the oxymatrine group， the protein expressions of PINK1， Parkin， and LC3-Ⅱ， apoptotic rate and the proportion of decrease in MMP were significantly decreased in PINK1 siRNA+oxymatrine group （

＜0.05）. Compared with the oxymatrine group， the protein expression of COX-2 in the COX-2+oxymatrine group was increased significantly （

＜0.05）， while the protein expressions of PINK1 and Parkin as well as the proportion of decrease in MMP were decreased significantly （

＜0.05）.

CONCLUSIONS

Oxymatrine can mediate the overactivity of mitophagy based on the PINK1/Parkin signaling pathway by inhibiting COX-2 expression， thus promoting the apoptosis of the MG63 osteosarcoma cell line.

关键词

氧化苦参碱骨肉瘤细胞凋亡线粒体自噬环氧合酶2PTEN诱导激酶1帕金森病蛋白2

Keywords

osteosarcomacell apoptosismitophagycyclooxygenase-2PTEN-induced putative kinase protein-1Parkinson disease protein-2

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