五味子乙素对谷氨酸诱导SH-SY5Y细胞损伤的保护作用及其机制

doi:10.13481/j.1671-587x.20160116

吉林大学学报(医学版) ›› 2016, Vol. 42 ›› Issue (01): 80-84.doi: 10.13481/j.1671-587x.20160116

五味子乙素对谷氨酸诱导SH-SY5Y细胞损伤的保护作用及其机制

杨擎, 隋欣, 王宝森, 于智莘, 朱凯, 律广富, 王建玮, 闫文昊, 张慧媛, 林嘉楠, 刘英娜, 李娜, 曲晓波

长春中医药大学研发中心药理实验室, 吉林长春 130117

收稿日期:2015-09-30 发布日期:2016-01-26
通讯作者: 李娜,副研究员(Tel:0431-86172151,E-mail:lhaln@hotmail.com);曲晓波,教授,博士研究生导师(Tel:0431-86172151,E-mail:quxiaobo0504@hotmail.com) E-mail:lhaln@hotmail.com;quxiaobo0504@hotmail.com
作者简介:杨擎(1989-),女,吉林省长春市人,在读中药学硕士,主要从事分子药理学方面的研究。
基金资助:
吉林省科技厅科技发展计划项目资助课题(20130522046JH);吉林省教育厅"十二五"科学技术研究项目资助课题(201484); 吉林省卫生厅青年科研基金资助课题(2013Q001);吉林省卫生计生委科研计划项目资助课题(2014Z024)

Protective effect of schizandra B on injury of SH-SY5Y cells induced by Glu and its mechanim

YANG Qing, SUI Xin, WANG Baosen, YU Zhixin, ZHU Kai, LYU Guangfu, WANG Jianwei, YAN Wenhao, ZHANG Huiyuan, LIN Jianan, LIU Yingna, LI Na, QU Xiaobo

Pharmacology Laboratory, Research and Development Center, Changchun University of Chinese Medicine, Changchun 130117, China

Received:2015-09-30 Published:2016-01-26

摘要/Abstract

摘要：

目的: 探讨五味子乙素(Sch B)对谷氨酸(Glu)致人神经母瘤细胞SH-SY5Y细胞损伤的保护作用,阐明其对ATR信号通路的影响。方法: 将处于对数生长期的SH-SY5Y细胞分为对照组、DMSO组、模型组、Sch B1组、Sch B2组和Sch B3组。对照组SH-SY5Y细胞不做任何处理;DMSO组SH-SY5Y细胞用17 mmol·L^-1 DMSO作用24h;模型组SH-SY5Y细胞用20 mmol·L^-1 Glu孵育24h造成细胞损伤模型;Sch B1、Sch B2和Sch B3组采用0.05、 0.10和1.00 μmol·L^-1浓度Sch B预处理SH-SY5Y细胞24 h后再用Glu(20 mmol·L^-1 )继续孵育24 h。MTT法检测细胞存活率,流式细胞术检测细胞周期变化,Western blotting法检测ATR、p-CHK1、p-Cdc25c和P53蛋白表达情况。结果: MTT法检测,与模型组比较,Sch B1、Sch B2和Sch B3组细胞存活率明显升高(P<0.05或P<0.01),且Sch B2组细胞存活率最高,故选取Sch B2组作为Sch B组进行后续实验。细胞周期检测,与对照组比较,模型组G₀/G₁期细胞所占百分比降低(P<0.05),S期和G₂/M 期细胞所占百分比升高(P<0.05);与模型组比较,Sch B组G₀/G₁期细胞所占百分比升高(P<0.05),S期和G₂/M 期细胞所占百分比降低(P<0.05)。Western blotting法检测,与对照组比较,模型组细胞中ATR、p-CHK1、p-Cdc25c和P53蛋白表达水平明显升高(P<0.05);与模型组比较,Sch B组细胞中ATR、p-CHK1、p-Cdc25c和P53蛋白表达水平均降低(P<0.05)。结论: Sch B对Glu致SH-SY5Y细胞损伤具有保护作用,其机制可能与下调ATR信号通路、减少细胞周期阻滞有关。

关键词: 五味子乙素, ATR信号通路, SH-SY5Y细胞, 谷氨酸

Abstract:

Objective: To investigate the protective effects of schizandra B (Sch B)on the injury of human neuroblastoma SH-SY5Y cells induced by glutamic acid (Glu), and to clarify its influence in ATR signaling pathway.Methods: The SH-SY5Y cells were divided into control group,dimethyl sulfoxide(DMSO)group,model group,Sch B1 group,Sch B2 group,and Sch B3 group.The SH-SY5Y cells in control group were untreated; the SH-SY5Y cells in DMSO group were treated with DMSO (17 mmol·L^-1)for 24 h;the SH-SY5Y cells in model group were treated with Glu (20 mmol·L^-1)for 24 h. The SH-SY5Y cells in Sch B1 group,Sch B2 group,and Sch B3 group were firstly pretreated with Sch B (0.1 μmol·L^-1)for 24 h,and exposed to 20 mmol·L^-1 Glu for 24 h at the same time.The survival rate of SH-SY5Y cells was detected by MTT assay,and the changes of cell cycle of SH-SY5Y cells were determined by flow cytometry.Western blotting method was used to detect the protein expressions.Results: The MTT detection results showed that compared with model group,the survival rates of SH-SY5Y cells in Sch B1 group, Sch B2 group,and Sch B3 group were increased (P<0.05 or P<0.01),and the survival rate in Sch B2 group was the highest (P<0.01),the Sch B2 group was selected as Sch B group for the following experiment.The cell cycle detection results showed that the percentage of SH-SY5Y cells in model group at G₀/G₁ phage was decreased(P<0.05); compared with control group,the percentages of SH-SY5Y cells in model group at S phage and G₂/M phage were increased (P<0.05);compared with model group, the percentage of SH-SY5Y cells in Sch B group at G₀/G₁ phage was increased (P<0.05),and the percentages of SH-SY5Y cells in Sch B group at S phage and G₂/M phage were decreased (P<0.05). The Western blotting detection results showed that compared with control group,the expression levels of ATR,p-CHK1,p-Cdc25c and P53 proteins in the SH-SY5Y cells in model group were increased (P<0.05);compared with model group,the expression levels of ATR,p-CHK1,p-Cdc25c and P53 proteins in the SH-SY5Y cell in Sch B group were decreased (P<0.05). Conclusion: Sch B could protect the damage of SH-SY5Y cells by GLu,and its mechanism may be related to inhibiting the ATR signaling pathway and reducing the cell cycle arrest.

Key words: schizandra B, ATR signaling pathway, SH-SY5Y cell, glutamic

中图分类号:

R285.5

杨擎, 隋欣, 王宝森, 于智莘, 朱凯, 律广富, 王建玮, 闫文昊, 张慧媛, 林嘉楠, 刘英娜, 李娜, 曲晓波. 五味子乙素对谷氨酸诱导SH-SY5Y细胞损伤的保护作用及其机制[J]. 吉林大学学报(医学版), 2016, 42(01): 80-84.

YANG Qing, SUI Xin, WANG Baosen, YU Zhixin, ZHU Kai, LYU Guangfu, WANG Jianwei, YAN Wenhao, ZHANG Huiyuan, LIN Jianan, LIU Yingna, LI Na, QU Xiaobo. Protective effect of schizandra B on injury of SH-SY5Y cells induced by Glu and its mechanim[J]. Journal of Jilin University Medicine Edition, 2016, 42(01): 80-84.

参考文献

[1] Zussy C,Brureau A,Keller E,et al.Alzheimer's disease related markers,cellular toxicity and behavioral deficits induced six weeks after oligomeric amyloid-β peptide injection in rats[J].PLoS One,2013,8(1):e53117.

[2] Xu MF,Xiong YY,Liu JK,et al.Asiatic acid,a pentacyclic triterpene in centella asiatica,attenuates glutamate-induced cognitive deficits in mice and apoptosis in SH-SY5Y cells[J].Acta Pharmacol Sinica,2012,33(5):578-587.

[3] Pivovarova NB,Andrews SB.Calcium-dependent mitochondrial function and dysfunction in neurons[J].FEBS J,2010,277(18):3622-3636.

[4] 黄丹华,周珊珊.氧化应激与阿尔茨海默病[J].国际神经病学神经外科学杂志,2012,39(3):268-270.

[5] 王佳丽,杨洪涛.五味子主要化学成分的药理研究[J].河南中医,2014,34(2):357-359.

[6] Chiu PY,Lam PY,Yan CW,et al.Schisandrin B protects against solar irradiation-induced oxidative injury in BJ human fibroblasts[J].Fitoterapia,2011,82(4):682-691.

[7] Liu W,Xu ZF.Research progress on antioxidants against neurotoxicity induced by methylmercury[J].Chin J Ind Med,2011,24(4):276-279.

[8] Zou Q,Yan MM,Cha YH,et al.Protective effect and probable mechanism of schisandrin B on PC12 cells[J].Herald Med,2013,32(8):1010-1014.

[9] 冯波,王蓉,盛树力.神经退行性疾病研究中拟神经细胞模型:人神经母细胞瘤株SH-SY5Y的来源特性及应用[J].中国临床康复,2006,10(6):121-123.

[10] Nampoothiri M,Reddy ND,John J,et al.Insulin blocks glutamate-induced neurotoxicity in differentiated SH-SY5Y neuronal cells[J].Behav Neurol,2014,2014:674164.doi:/10.1155/2014/674164.

[11] 邹琼,严明敏,查运红,等.五味子乙素保护神经细胞作用及其可能机制[J].医药导报,2013,32(8):1010-1014.

[12] 梁婧,侯海燕,兰晓霞,等.五味子乙素的药理作用及其分子机制的研究进展[J].中国现代应用药学,2014,31(4):506-510.

[13] 孙佳,李胜范,郑丽丽.ATM/ATR在DNA损伤反应中的作用[J].中华临床医师杂志:电子版,2011,5(6):1683-1686.

[14] Brooks CL,Gu W.New insights into p53 activation[J].Cell Res,2010,20(6):614-621.

[15] Zilfou JT,Lowe SW.Tumor suppressive functions of p53[J].Cold Spring Harb Perspect Biol,2009,1(5):a001883.

五味子乙素对谷氨酸诱导SH-SY5Y细胞损伤的保护作用及其机制

Protective effect of schizandra B on injury of SH-SY5Y cells induced by Glu and its mechanim

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