| 王 侃,程 志,吕刚飞,刘志飞,张 蕾,刘星汝.基于肝癌细胞线粒体功能受损和caspase-3信号通路探讨罗哌卡因促进肝癌细胞凋亡的作用机制研究[J].,2023,(2):244-250 |
| 基于肝癌细胞线粒体功能受损和caspase-3信号通路探讨罗哌卡因促进肝癌细胞凋亡的作用机制研究 |
| To Investigate the Mechanism of Ropivacaine Promoting Apoptosis of Hepatocellular Carcinoma Cells Based on Mitochondrial Function Impairment and Caspase-3 Signaling Pathway |
| 投稿时间：2022-05-23 修订日期：2022-06-18 |
| DOI：10.13241/j.cnki.pmb.2023.02.008 |
| 中文关键词: 罗哌卡因 肝癌细胞 线粒体 caspase-3 凋亡 |
| 英文关键词: Ropivacaine Hepatoma carcinoma cell Mitochondria Caspase-3 Apoptosis |
| 基金项目:山东省医药卫生科技发展计划项目（2017DX0355） |
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| 中文摘要: |
| 摘要 目的：基于肝癌细胞线粒体功能受损和天冬氨酸蛋白水解酶3（caspase-3）信号通路探讨罗哌卡因促进肝癌细胞凋亡的作用机制。方法：选用细胞株人肝癌细胞BEL-7402进行实验研究。用不同浓度罗哌卡因处理BEL-7402细胞后，采用溴化噻唑蓝四氮唑（MTT）法检测肝癌细胞的增殖情况，光镜及4，6-二苯胺-2-苯吲哚二盐酸盐（DAPI）溶液染色观察细胞形态，台盼蓝染色法测定细胞活力，流式细胞术分析BEL-7402细胞的凋亡情况，电子显微镜下观察细胞线粒体，激光共聚焦显微镜观察caspase-3在BEL-7402细胞中的细胞核迁移情况，蛋白免疫印迹试验评价罗哌卡因对细胞质凋亡相关蛋白、线粒体凋亡相关蛋白、BEL-7402细胞和线粒体凋亡相关蛋白表达的影响。结果：罗哌卡因能够抑制肝癌细胞的生长，并呈剂量依赖性和时间依赖性。罗哌卡因可诱导BEL-7402细胞发生凋亡，显著增加BEL-7402细胞的凋亡率。罗哌卡因能够损伤肝癌细胞线粒体功能。激光共聚焦显微镜观察显示caspase-3分子迁移到细胞核。罗哌卡因与caspase-3相互作用，促进caspase-3向细胞核内迁移，刺激caspase-3和聚腺苷二磷酸核糖聚合酶（PARP-1）、天冬氨酸蛋白水解酶9（caspase-9）蛋白的表达，抑制B细胞淋巴瘤-2基因（Bcl-2）的表达，促进凋亡酶激活因子（Apaf-1）的表达，促进线粒体释放细胞色素C（Cytochrome C），激活caspase-3活性。结论：罗哌卡因具有促进肝癌细胞凋亡的作用，其作用机制可能与破坏肝癌细胞线粒体功能和激活caspase-3信号通路有关。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the mechanism of ropivacaine promoting apoptosis of hepatoma carcinoma cell based on mitochondrial dysfunction and caspase-3 signaling pathway. Methods: Human hepatoma carcinoma cell line BEL-7402 was used for experimental study. After Bel-7402 cells were treated with different concentrations of ropivacaine, the proliferation of hepatoma carcinoma cell was detected by MTT method, the cell morphology was observed by light microscopy and DAPI solution staining, the cell viability was determined by trypan blue staining, the apoptosis of Bel-7402 cells was analyzed by flow cytometry, and the mitochondria were observed under electron microscopy. The nuclear migration of caspase-3 in Bel-7402 cells was observed by laser confocal microscopy, and the effects of ropivacaine on the expression of cytoplasmic apoptosis-related proteins, mitochondrial apoptosis-related proteins, Bel-7402 cells and mitochondrial apoptosis-related proteins were evaluated by western blot assay. Results: Ropivacaine can inhibit the growth of hepatoma carcinoma cell in a dose-dependent and time-dependent manner. The apoptosis of Bel-7402 cells was induced by ropivacaine, and the apoptosis rate of Bel-7402 cells was significantly increased. Ropivacaine can injure mitochondrial function of hepatoma carcinoma cells. Laser confocal microscopy showed the migration of caspase-3 molecules to the nucleus. Ropivacaine interacts with caspase-3 to promote the migration of caspase-3 into the nucleus, stimulate the expression of caspase-3, PARP-1 and caspase-9 proteins, inhibit the expression of Bcl-2, promote the expression of APAF-1, promote mitochondria to release Cytochrome C and activate caspase-3 activity. Conclusion: Ropivacaine can promote the apoptosis of hepatoma carcinoma cell, and its mechanism may be related to the destruction of mitochondrial function and activation of caspase-3 signaling pathway. |
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