尼氟灭酸对氯化钴诱导海马神经元凋亡的抑制作用

doi:10.13481/j.1671-587x.20150506

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (05): 914-918.doi: 10.13481/j.1671-587x.20150506

尼氟灭酸对氯化钴诱导海马神经元凋亡的抑制作用

田晶¹, 邹荣军¹, 梁祖凤¹, 石婉婷¹, 齐玲², 李妍³

1. 吉林医药学院生理学教研室, 吉林吉林 132013;
2. 吉林医药学院病理学教研室, 吉林吉林 132013;
3. 吉林医药学院病原学教研室, 吉林吉林 132013

收稿日期:2015-01-08 出版日期:2015-09-28 发布日期:2015-09-29
通讯作者: 齐玲,教授(Tel:0432-64560027,E-mail:qiling17187@163.com) E-mail:qiling17187@163.com
作者简介:田晶(1973-),女,吉林省吉林市人,副教授,医学硕士,主要从事脑肿瘤及脑缺血损伤方面的研究。
基金资助:
国家自然科学基金资助课题(81201671);吉林省教育厅科研项目资助课题(2014369)

Inhibitory effects of niflumic acid on apoptosis induced by cobalt chloride in hippocampus neurons

TIAN Jing¹, ZOU Rongjun¹, LIANG Zufeng¹, SHI Wanting¹, QI Ling², LI Yan³

1. Department of Physiology, Jilin Medical College, Jilin 132013, China;
2. Department of Pathology, Jilin Medical College, Jilin 132013, China;
3. Department of Pathogenic Biology, Jilin Medical College, Jilin 132013, China

Received:2015-01-08 Online:2015-09-28 Published:2015-09-29

摘要/Abstract

摘要：

目的:探讨尼氟灭酸(NFA)对氯化钴(CoCl₂)诱导的海马神经元凋亡的抑制作用,阐明其可能的机制。方法:原代培养乳鼠海马细胞,并在倒置显微镜下观察海马细胞的形态变化。将海马神经元随机分为对照组(含1%血清培养液)、CoCl₂损伤组(含200 μmol·L^-1NFA等体积培养液)和不同浓度NFA组(在CoCl₂损伤组基础上加入20、40、80 μmol·L^-1 NFA)。MTT法检测各组海马神经元的存活率,流式细胞术分析海马神经元调亡率, ELISA法检测神经元培养液上清中相关凋亡蛋白Bcl-2、Bax、caspase-3和缺氧相关蛋白低氧诱导因子(HIF-1α)的表达水平。结果:海马神经元在培养7 d后形态规则,突触相互交织形成网络。MTT检测,与对照组比较,CoCl₂损伤组神经元存活率明显降低(P<0.01);与CoCl₂损伤组比较,20、40和80 μmol·L^-1NFA组神经元存活率升高(P<0.05或P<0.01)。流式细胞术检测,与对照组比较,CoCl₂损伤组神经元早期凋亡率、晚期凋亡率和总凋亡率升高(P<0.05或P<0.01);与CoCl₂损伤组比较,20、40和80 μmol·L^-1NFA组神经元早期凋亡率、晚期凋亡率及总凋亡率下降 (P<0.05或P<0.01)。ELISA法检测,与对照组比较,CoCl₂损伤组神经元培养液上清中HIF-1α、caspase-3 和Bax表达水平升高(P<0.01);与CoCl₂损伤组比较,20和40 μmol·L^-1 NFA组神经元培养上清中HIF-1α、caspace-3 和Bax表达水平下降,Bcl-2表达水平升高(P<0.05或P<0.01)。结论:NFA可以降低CoCl₂对海马神经元的缺氧损伤作用,其机制与抑制HIF-1α的表达、下调Bax/Bcl-2及抑制神经元凋亡有关联。

关键词: 尼氟灭酸, 海马神经元, 氯化钴, 细胞凋亡

Abstract:

Objective To explore the inhibitory effects of niflumic acid (NFA) on the apoptosis of hippocampal neurons induced by cobalt chloride (CoCl₂),and to elucidate the possible mechanisms. Methods The hippocampal neurons of rats were isolated and cultured,and inverted phase contrast microscope was used to observe the morphological changes of neurons.The hippocampal neurons were cultured and randomly divided into control group (1% serum medium),CoCl₂ injury group (200 μmol·L^-1 CoCl₂) and different concentrations(20,40,80 μmol·L^-1)of NFA groups.The survival rates of hippocampal neurons in various groups were measured by MTT assay.The apoptotic rates of hippocampal neurons were analyzed by flow cytometry.ELISA method was performed to observe the expression levels of apoptosis associated proteins Bax,Bcl-2,caspase-3 and hypoxia-inducible factor 1α (HIF-1α) in the supernatant of neurons. Results The hippocampal neurons showed regular shapes and the synapses reticulated after cultured for 7 d. Compared with control group,the survival rate of neurons in CoCl₂ injury group was decreased (P<0.01).Compared with CoCl₂ injury group,the survival rates of neurons in 20,40,and 80 μmol·L^-1 NFA groups were increased significantly (P<0.05 or P<0.01).The results of flow cytometry showed that compared with control group,the apoptotic rate in CoCl₂ injury group was increased (P<0.05);compared with CoCl₂ injury group,the apoptotic rates of neurons in 20,40,and 80 μmol·L^-1 NFA groups were decreased (P<0.05 or P<0.01).The ELISA results showed that the expression levels of HIF-1α,caspase-3,and Bax in the supernatant of neurons in CoCl₂ injury group were increased compared with control group(P<0.01);compared with CoCl₂ injury group,the expression levels of HIF-1α,caspase-3,and Bax in the supernatant of neurons in 20 and 40 μmol·L^-1 NFA groups were decreased,while the expression level of Bcl-2 was increased (P<0.05 or P<0.01). Conclusion NFA can protect the hippocampal neurons from hypoxia injury induced by CoCl₂,which may be related to suppressing the expression of HIF-1α,reducing the ratio of Bax/Bcl2 and suppressing the apoptosis of neurons.

Key words: niflumic acid, hippocampal neurons, cobalt chloride, apoptosis

中图分类号:

R737.33

田晶, 邹荣军, 梁祖凤, 石婉婷, 齐玲, 李妍. 尼氟灭酸对氯化钴诱导海马神经元凋亡的抑制作用[J]. 吉林大学学报(医学版), 2015, 41(05): 914-918.

TIAN Jing, ZOU Rongjun, LIANG Zufeng, SHI Wanting, QI Ling, LI Yan. Inhibitory effects of niflumic acid on apoptosis induced by cobalt chloride in hippocampus neurons[J]. Journal of Jilin University Medicine Edition, 2015, 41(05): 914-918.

参考文献

[1] Abouzid KA,Khalil NA,Ahmed EM,et al.Synthesis and biological evaluation of new heteroaryl carboxylic acid derivatives as anti-inflammatory-analgesic agents [J].Chem Pharm Bull (Tokyo),2013,61(2):222-228.

[2] Srtuass KI,Marini AM.Cyelooxygenase-2 ihnbitiion protects cultured cerebellar granule neurons from glutamate-mediated cell death [J].J Neurotrauma,2002,19(5):627-638.

[3] Feng ZH,Wang TG,Li DD,et al.Cyclooxygenase-2-deficient mice are resistant to 1-methyl-4-phenyl1,2,3,6-tetrahydropyridine-induced damage of dopaminergic neurons in the substantia nigra [J].Neurosci Lett,2002,329(3):354-358.

[4] 田晶,陶凌,曹云新,等.尼氟灭酸对肝癌细胞增殖的影响 [J].生理学报,2003,55(2):160-164.

[5] Jiang J,Borisenko GG,Osipov A,et al.Arachidonic acid-induced carbon-centered radicals and phospholipid peroxidation in cyclo-oxygenase-2-transfected PC12 cells [J].J Neurochem,2004,90(5):1036-1049.

[6] Chavez JC,LaManna JC.Activation of hypoxia-inducible factor-1 in the rat cerebral cortex after transient global ischemia:potential role of insulin-like growth factor-1 [J].J Neurosci,2002,22(20):8922-8931.

[7] Gilkes DM,Semenza GL,Wirtz D.Hypoxia and the extracellular matrix:drivers of tumour metastasis [J].Nat Rev Cancer,2014,14(6):430-439.

[8] Wang LX,Zeng JP,Wei XB,et al.Effects of scutellarin on apoptosis induced by cobalt chloride in PC12 cells [J].Chin J Physiol,2007,50(6):301-307.

[9] Dorsey ER,George BP,Leff B,et al.The coming crisis:obtaining carefor the growing burden of neurodegenerative condition [J].Neurology,2013,80(21):1989-1996.

[10] Mehta SL,Manhas N,Raghubir R.Molecular targets in cerebral ischemia for developing novel therapeutics [J].Brain Res Rev,2007,54(1):34-66.

[11] Igarashi KM,Ito HT,Moser EI,et al.Functional diversity along the transverse axis of hippocampal area CA1 [J].FEBS Lett,2014,588(15):2470-2476.

[12] Zhang Y,Zhang H,Feustel PJ,et al.DCPIB,a specific inhibitor of volume regulated anion channels,reduces infarct size in MCAo and the release of glutamate in the ischemic cortical penumbra [J].Exp Neurol,2008,210(2):514-520.

[13] Semenza G.Signal transduction to hypoxia-inducible factor 1 [J].Biochem Pharmacol,2002,64(5-6):993-998.

[14] Liu BN,Han BX,Liu F.Neuroprotective effect of pAkt and HIF-1 α on ischemia rats [J].Asian Pac J Trop Med,2014,7(3):221-225.

[15] Ceradini DJ,Yao D,Grogan RH,et al.Decreasing intracellular superoxide corrects defective ischemia-induced new vessel formation in diabetic mice [J].Biol Chem,2008,283(16):10930-10938.

[16] Althaus J,Bernaudin M,Petit E,et al.Expression of the gene encoding thepro-apoptotic BNIP3 protein and stimulation of hypoxia-inducible factor-1 alpha (HIF-1 alpha) protein following focal cerebral ischemia in rats [J].Neurochem Int,2006,48(8):687-695.

[17] Yu CH,Moon CT,Sur JH,et al.Serial expression of hypoxia inducible factor-1α and neuronal apoptosis in hippocampus of rats with chronic ischemic brain [J].J Korean Neurosurg Soc,2011,50(6):481-485.

[18] 柯跃斌,郑荣梁.自由基毒理学 [M].北京:人民卫生出版社,2012:218-228.

[19] Larkins TL,Nowell M,Singh S,et al.Inhibition of cyclooxygenase-2 decreases breast cancer cell motility,invasion and matrix metalloproteinase expression [J].BMC Cancer,2006,10(6):181.

[20] Kim BM,Maeng K,Lee KH,et al.Combined treatment with the Cox-2 inhibitor niflumic acid and PPARγ ligand ciglitazone induces ER stress/caspase-8-mediated apoptosis in human lung cancer cells [J].Cancer Lett,2011,300(2):134-144.

[21] 杜森,夏青波,叶琳,等.大鼠海马认知障碍对糖脂代谢的影响及其与海马神经元凋亡的关系[J].中国老年学杂志,2014,35(4):1000-1001.

尼氟灭酸对氯化钴诱导海马神经元凋亡的抑制作用

Inhibitory effects of niflumic acid on apoptosis induced by cobalt chloride in hippocampus neurons

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