异氟醚对Aβ25-35诱导大鼠PC12细胞自噬和凋亡的影响

doi:10.7694/jldxyxb20140112

吉林大学学报(医学版)

异氟醚对Aβ25-35诱导大鼠PC12细胞自噬和凋亡的影响

王伟^1,2，朴美花¹，王艳姝¹，刘楠¹，岳云³，冯春生¹

（1. 吉林大学第一医院麻醉科，吉林长春 130021；2. 吉林省长春市传染病医院麻醉科，吉林长春 130123；3. 首都医科大学附属北京朝阳医院麻醉科，北京 100020）

收稿日期:2013-10-11 出版日期:2014-01-28 发布日期:2014-01-28
通讯作者: 冯春生 E-mail: (Tel：0431-88782955，E-mail：fcs1971@hotmail.com)
作者简介:王伟(1979-)，男，吉林省长春市人，主治医师，在读医学硕士，主要从事全麻药对阿尔茨海默病发病机制影响的研究。
基金资助:
国家自然科学基金资助课题（81141065,81271215）

Effects of isoflurane on autophagy and apoptosis induced by Aβ25-35 in rat PC12 cells

WANG Wei^{1 ,2},PIAO Mei-hua¹,WANG Yan-shu¹,LIU Nan¹,YUE Yun³,FENG Chun-sheng¹

(1. Department of Anesthesiology,First Hospital,Jilin University,Changchun 130021,China;2. Department of Anesthesiology,Changchun Infectious Diseases Hospital,Changchun 130123,China;3. Department of Anesthesiology,Affiliated Beijing Chaoyang Hospital, Capital University of Medical Sciences,Beijing 100020,China)

Received:2013-10-11 Online:2014-01-28 Published:2014-01-28

摘要/Abstract

摘要：

目的：探讨吸入麻醉药异氟醚对β淀粉样蛋白（Aβ）25-35诱导的大鼠PC12细胞自噬和凋亡的影响，阐明其可能的作用机制。方法：将PC12细胞随机分为正常对照组（C组）、10 μmol•L-1Aβ25-35 处理组（Aβ组）、2%异氟醚处理组（Iso组）和2%异氟醚与10 μmol•L-1Aβ25-35联合处理组（Iso+Aβ组）。各组PC12细胞药物处理6 h后应用透射电镜观察自噬体，Western blotting法检测自噬相关蛋白LC3-Ⅱ、p62和自噬信号调控基因p-mTOR、Beclin-1表达；药物处理24 h后应用四甲基偶氮唑盐（MTT）比色法检测细胞存活率，Western blotting法检测凋亡相关蛋白Bcl-2和caspase-3表达。结果：与C组比较，Aβ组PC12细胞中可见大量的自噬体，LC3-Ⅱ、Beclin-1和caspase-3表达量升高（P<0.05），p62、p-mTOR和Bcl-2表达量降低（P<0.05），细胞存活率降低（P<0.05）;Iso组PCI2细胞未见自噬体，LC3-Ⅱ
表达量降低（P<0.05），p62、p-mTOR和caspase-3表达量升高（P<0.05），细胞存活率降低（P<0.05）;与Aβ组比较，Iso+Aβ组PC12细胞仅见少量自噬体，LC3-Ⅱ表达量降低（P<0.05），p62、p-mTOR、Beclin-1和caspase-3表达量升高（P<0.05），细胞存活率降低（P<0.05）,Bcl-2表达无明显变化（P>0.05）。结论：异氟醚能够抑制Aβ25-35诱导大鼠PC12细胞自噬的活化，促进凋亡蛋白的表达，其作用机制与激活自噬信号调控基因mTOR、抑制Beclin-1的表达有关。

关键词: 异氟醚, 阿尔茨海默病, PC12细胞, 自噬, 细胞凋亡, &beta, 淀粉样蛋白

Abstract:

To explore the effects of isoflurane on autophagy and apoptosis of rat PC12 cells induced by Aβ25-35,and to elucidate the possible mechanisms.Methods PC12 cells were randomly divided into 4 groups (n=6).Control group:the PC12 cells were administered with normal medium;10 μmol•L-1 Aβ25-35 group(Aβ group):the PC12 cells were administered with 10 μmol•L-1 Aβ25-35;2% isoflurane group(Iso group):the PC12 cells were administered with 2% isoflurane;2% isoflurane and 10 μmol•L-1 Aβ25-35 group(Iso+Aβ group):the PC12 cells were administered with 2% isoflurane +10 μmol•L-1 Aβ25-35.The autophagosomes of PC12 cells in various groups were examined by transmission electron microscope,and the expressions of autophagy associated protein LC3-Ⅱ,p62 and autophagic signal gene mTOR,Beclin-1 were detected by Western blotting method 6 h after administration;the cell viability was determined by MTT assay,and Western blotting method was performed to observe the expressions of apoptosis associated protein Bcl-2 and caspase-3 24 h after administration.Results Compared with control group,great amounts of autophagosomes were presented in PC12 cells in Aβ group,the expressions of LC3-Ⅱ,Beclin-1 and caspase-3 were significantly increased (P<0.05),and the expressions of p62,p-mTOR and Bcl-2 were significantly decreased as well as the survival rate (P<0.05);however,autophagosomes failed to be presented in PC12 cells in Iso group,the expressions of LC3-Ⅱ,Beclin-1 and Bcl-2 as well as cell survival rate were significantly decreased (P<0.05),and the expression of p62,p-mTOR and caspase-3 were significantly increased(P<0.05).Compared with Aβ group,only a few autophagosomes were found in PC12 cells in Iso+Aβ group,the expressions of LC3-Ⅱ and Beclin-1 as well as the survival rate were significantly decreased (P<0.05),and the expressions of p62,p-mTOR and caspase-3 were significantly increased(P<0.05),but no changes occurred with the expression of Bcl-2 (P>0.05).Conclusion Isoflurane can inhibit autophagy of rat PC12 cells induced by Aβ25-35 and aggravate the expressions of apoptosis rrelated proteins through activation of autophagic signal
gene mTOR and inhibition of Beclin-1.

Key words: isoflurane, Alzheimer&rsquo, s disease, PC12 cells, autophagy, apoptosis, beta-amyloid protein

中图分类号:

R614.24

王伟，朴美花，王艳姝，刘楠，岳云，冯春生. 异氟醚对Aβ25-35诱导大鼠PC12细胞自噬和凋亡的影响[J]. 吉林大学学报(医学版), 2014, 40(01): 49-54.

WANG Wei,PIAO Mei-hua,WANG Yan-shu,LIU Nan,YUE Yun,FENG Chun-sheng. Effects of isoflurane on autophagy and apoptosis induced by Aβ25-35 in rat PC12 cells[J]. Journal of Jilin University Medicine Edition, 2014, 40(01): 49-54.

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异氟醚对Aβ25-35诱导大鼠PC12细胞自噬和凋亡的影响

Effects of isoflurane on autophagy and apoptosis induced by Aβ25-35 in rat PC12 cells

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