贵州医科大学学报

2020, v.45;No.235(04) 373-378+386

[打印本页] [关闭]
本期目录(Current Issue) | 过刊浏览(Past Issue) | 高级检索(Advanced Search)

α-硫辛酸对缺血再灌注诱导的PC12细胞应激性凋亡抑制作用的研究
Study of Inhibitory Effect of α-Lipoic Acid on Apoptosis of PC12 Cells Induced by Ischemia-Reperfusion

谢鹏;任真奎;吕菊;胡玉梅;吴昌学;禹文峰;
XIE Peng;REN Zhenkui;LV Jv;HU Yumei;WU Changxue;YU Wenfeng;Key Laboratory of Molecular Biology of Guizhou Medical University;Education Ministry Key Laboratory of Endemic and Minority Diseases of Guizhou Medical University;Department of Laboratory,People's Hospital of Southwest Guizhou Autonomous Prefecture;

摘要(Abstract):

目的:探讨α-硫辛酸(α-LA)抑制缺血再灌注诱导的大鼠肾上腺嗜铬神经瘤细胞(PC12)内质网应激性凋亡的分子机制。方法:传代培养PC12细胞分为正常组(Control组)、氧糖剥夺/再灌注(OGD/R) 4 h组(OGD4组)、OGD 4 h后复氧不同时间段组(OGD4+R6、OGD4+R12、OGD4+R18以及OGD4+R24组)、在OGD4 h后复氧阶段给予不同浓度(0. 01、0. 05、0. 1、0. 25、0. 5、1、2、5和10 mmol/L)α-LA共同复氧培养24 h作为α-LA处理组(OGD/R+α-LA组); Control组细胞在完全培养基、正常氧培养箱中培养,OGD/R组和OGD/R+α-LA组细胞按相应方法进行培养处理;蛋白免疫印迹法检测各组PC12细胞中内质网应激[葡萄糖调节蛋白78(GRP78)、环磷酸腺苷反应元件结合转录因子同源蛋白(CHOP)]和凋亡[B淋巴细胞瘤-2相关X (Bax)、裂解型胱天蛋白水解酶3(Cleaved-caspase3)]相关分子的表达;细胞活性检测试剂盒-8(CCK-8试剂盒)检测各组PC12细胞的存活率,分析α-LA作用的有效工作浓度。结果:与Control组比较,OGD4 h后复氧不同时间段均诱导了PC12细胞发生内质网应激性的凋亡,表现为内质网应激相关分子GRP78、CHOP和凋亡蛋白Bax、Cleavedcaspase3的表达伴随复氧时间段的延长而逐渐升高(P <0. 05),在OGD4+R24组升高最明显(P <0. 05);与Control组比较,OGD4+R24组细胞的存活率明显下降(P <0. 05);与OGD4+R24组比较,OGD/R+α-LA组α-LA在0. 05~5 mmol/L浓度范围内增加了细胞的存活率(P <0. 05),0. 5 mmol/L时细胞存活率升高最明显(P <0. 05),10 mmol/L时细胞存活率明显降低(P <0. 05);与OGD4+R24组比较,OGD/R+α-LA(0. 5 mmol/L)组凋亡分子Bax、Cleaved-caspase3以及内质网应激相关分子GRP78、CHOP表达明显降低(P <0. 05)。结论:缺血再灌注诱导了PC12细胞发生内质网应激性的凋亡,α-LA能够降低缺血再灌注诱导的PC12损伤,其机制可能与降低了缺血再灌注诱导的内质网应激性凋亡有关。
Objective: To explore the molecular mechanism by which α-lipoic acid( α-LA) inhibits endoplasmic reticulum( ER) stress-mediated apoptosis in rat adrenal pheochromocytoma cells( PC12)induced by ischemia-reperfusion. Methods: PC12 cells were cultured under normal condition( control group),oxygen-sugar deprivation/reperfusion( OGD/R) 4 h group( OGD4 group),and OGD4 +reoxygenation( R) at different time periods( OGD4 + R6,OGD4 + R12,OGD4 + R18,and OGD4 +R24 groups). During rexoygenation,OGD4/R were then given α-LA at different concentrations( 0. 01,0. 05,0. 1,0. 25,0. 5,1,2,5 and 10 mmol/L) for 24 h as OGD/R + α-LA group.Western blot was used to detect glucose-regulated protein 78( GRP78),cyclic adenosine phosphate response element binding transcription factor homolog protein( CHOP),Bax and Cleaved-caspase3.Cell viability was detected using CCK-8 kit. Results: Compared with the control group,reoxygenation induced PC12 cells to undergo ER-mediated apoptosis in OGD4 group,and the expression levels of GRP78,CHOP,Bax and Cleaved-caspase3 gradually increased over the prolongation of reoxygenation period( P < 0. 05),and reached the highest levels in OGD4 + R24 group( P < 0. 05). When compared with the control group,the cell survival rate was remarkably decreased in the OGD4 + R24 group( P < 0. 05) . α-LA increased the cell survival rates at the concentration range of 0. 05 ~5 mmol/L in OGD/R + α-LA group( P < 0. 05). The survival rate was the highest level at 0. 5 mmol/L of α-LA,but reduced at 10 mmol/L of α-LA( P < 0. 05). When compared with the OGD4 + R24 group,the expression levels of Bax,Cleaved-caspase3,GRP78 and CHOP were significantly reduced in OGD/R + α-LA( 0. 5 mmol/L)( P < 0. 05). Conclusion: Ischemia-reperfusion induces endoplasmic reticulum stress-mediated apoptosis in PC12 cells. α-LA can reduce ischemia-reperfusion-induced cell injury by downregulating GRP78 and CHOP.

关键词(KeyWords): 再灌注损伤;大鼠;α-硫辛酸;肾上腺嗜铬神经瘤细胞;细胞损伤;氧糖剥夺/再灌注;内质网压力介导的凋亡
reperfusion injury;rats;alpha-lipoic acid(α-LA);adrenal pheochromocytoma cell;cellular injury;oxygen-sugar deprivation/reperfusion;ER stress-mediated apoptosis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81360199);; 黔科中引地[(2019)4008号];; 贵州省卫生健康委科学技术基金(gzwjkj2019-1-039);; 黔西南州科技局基金(2019-1-10)

作者(Author): 谢鹏;任真奎;吕菊;胡玉梅;吴昌学;禹文峰;
XIE Peng;REN Zhenkui;LV Jv;HU Yumei;WU Changxue;YU Wenfeng;Key Laboratory of Molecular Biology of Guizhou Medical University;Education Ministry Key Laboratory of Endemic and Minority Diseases of Guizhou Medical University;Department of Laboratory,People's Hospital of Southwest Guizhou Autonomous Prefecture;

Email:

DOI: 10.19367/j.cnki.1000-2707.2020.04.001

参考文献(References):

文章评论(Comment):

序号(No.) 时间(Time) 反馈人(User) 邮箱(Email) 标题(Title) 内容(Content)
反馈人(User) 邮箱地址(Email)
反馈标题(Title)
反馈内容(Content)
扩展功能
本文信息
服务与反馈
本文关键词相关文章
本文作者相关文章
中国知网
分享