氧糖剥夺(OGD)诱导SH-SY5Y细胞自噬性死亡
摘要
目的探讨应用体外氧糖剥夺(Oxygen-glucose Deprivation,OGD)模型诱导人神经母细胞瘤SH-SY5Y细胞自噬性死亡与自噬过程的潜在作用机制。方法应用自噬抑制剂(3-Methyladenine,3MA)和siRNA沉默ATG5的表达后,OGD处理24小时,LDH检测各组细胞的死亡率(P<0.01);Western blotting检测各组细胞中自噬相关蛋白的表达水平。结果 OGD诱导SH-SY5Y细胞自噬水平升高;细胞死亡率增加,应用3MA和siRNA技术沉默ULK1基因后,细胞死亡率(P<0.05)及自噬蛋白Beclin1、LC3蛋白表达水平显著降低,P62蛋白表达水平升高。结论自噬参与并促进了OGD诱导SH-SY5Y细胞死亡。
引文
[1]Lee JC,Won MH,Neuroprotection of antioxidant enzymes against transient global cerebral ischemia in gerbils[J].Anat Cell Biol,2014,47(3):149-56.
[2]Jauch EC,Saver JL,Adams HP,et al.American Heart Association Stroke Council,Council on Cardiovascular Nursing,Council on Peripheral Vascular Disease,Council on Clinical Cardiology.Guidelines for the early management of patients with acute ischemic stroke:a guideline for healthcare professionals from the American Heart Association/American Stroke Association[J].Stroke,2013,44:870-947.
[3]Molina CA,Alvarez-Sabín J.Recanalization and reperfusion therapies for acute ischemic stroke[J].Cerebrovasc Dis,2009,27:S162-S167.
[4]Adeoye O.,Hornung R.,Khatri P.,Kleindorfer D.Recombinant tissue-type plasminogen activator use for ischemic stroke in the United States:a doubling of treatment rates over the course of 5 years[J].Stroke,2011,42(7):1952-1955.
[5]Chapman S.N.,Mehndiratta P.,Johansen M.C.,McMurry T.,Johnston K.C.,Southerland A.M.Current perspectives on the use of intravenous recombinant tissue plasminogen activator(tPA)for treatment of acute ischemic stroke[J].Vascular Health and Risk Management,2014,10:75-87.
[6]Adibhatla R.,Hatcher J.Tissue plasminogen activator(tPA)and matrix metalloproteinases in the pathogenesis of stroke:therapeutic strategies.CNS&Neurological Disorders Drug Targets[J],2008,7(3):243-253.
[7]Mari?o G,Madeo F,Kroemer G,et al Autophagy for tissue homeostasis and neuroprotection[J].Curr Opin Cell Biol,2011,23:198-206.
[8]Lipinski,M.M.et al.Genome-wide analysis reveals mechanisms modulating autophagy in normal brain aging and in Alzheimer’s disease[J].Proceedings of the National Academy of Sciences of the United States of America,2012,107,14164-14169.
[9]Ravikumar,B.et al.Inhibition of mTOR induces autophagy and reduces toxicity of polyglutamine expansions in fly and mouse models of Huntington disease[J].Nature genetics,2011,36,585-595.
[10]Mizushima,N.,Yoshimori,T.&Ohsumi,Y.Role of the Apg12 conjugation system in mammalian autophagy[J].The international journal of biochemistry&cell biology,2003,35,553-561.
[2]Jauch EC,Saver JL,Adams HP,et al.American Heart Association Stroke Council,Council on Cardiovascular Nursing,Council on Peripheral Vascular Disease,Council on Clinical Cardiology.Guidelines for the early management of patients with acute ischemic stroke:a guideline for healthcare professionals from the American Heart Association/American Stroke Association[J].Stroke,2013,44:870-947.
[3]Molina CA,Alvarez-Sabín J.Recanalization and reperfusion therapies for acute ischemic stroke[J].Cerebrovasc Dis,2009,27:S162-S167.
[4]Adeoye O.,Hornung R.,Khatri P.,Kleindorfer D.Recombinant tissue-type plasminogen activator use for ischemic stroke in the United States:a doubling of treatment rates over the course of 5 years[J].Stroke,2011,42(7):1952-1955.
[5]Chapman S.N.,Mehndiratta P.,Johansen M.C.,McMurry T.,Johnston K.C.,Southerland A.M.Current perspectives on the use of intravenous recombinant tissue plasminogen activator(tPA)for treatment of acute ischemic stroke[J].Vascular Health and Risk Management,2014,10:75-87.
[6]Adibhatla R.,Hatcher J.Tissue plasminogen activator(tPA)and matrix metalloproteinases in the pathogenesis of stroke:therapeutic strategies.CNS&Neurological Disorders Drug Targets[J],2008,7(3):243-253.
[7]Mari?o G,Madeo F,Kroemer G,et al Autophagy for tissue homeostasis and neuroprotection[J].Curr Opin Cell Biol,2011,23:198-206.
[8]Lipinski,M.M.et al.Genome-wide analysis reveals mechanisms modulating autophagy in normal brain aging and in Alzheimer’s disease[J].Proceedings of the National Academy of Sciences of the United States of America,2012,107,14164-14169.
[9]Ravikumar,B.et al.Inhibition of mTOR induces autophagy and reduces toxicity of polyglutamine expansions in fly and mouse models of Huntington disease[J].Nature genetics,2011,36,585-595.
[10]Mizushima,N.,Yoshimori,T.&Ohsumi,Y.Role of the Apg12 conjugation system in mammalian autophagy[J].The international journal of biochemistry&cell biology,2003,35,553-561.