一氧化氮诱导星形胶质细胞HIF-1α活化的实验研究  

A Experimental Study on Activation of HIF-1αin Astrocytes Induced by Nitric Oxide

作　　者：董越[1] 王朝驹[1] 王德伟 王黎[3] 李凯 伏建峰[3] 史清海[3] DONG Yue;WANG Zhao-ju;WANG De-wei;WANG Li;LI Kai;FU Jian-feng;SHI Qing-hai(The Second Department of Cadre Ward,1.Traditional Chinese Medicine Hospital Affiliated to Xinjiang Medical University,Urumqi 830000,China;Department of Rehabilitation Medicine,1.Traditional Chinese Medicine Hospital Affiliated to Xinjiang Medical University,Urumqi 830000,China;Clinical Laboratory Diagnostic Center,General Hospital of Xinjiang Military Area Command,Urumqi 830000,China)

机构地区：[1]新疆医科大学附属中医医院干二科,乌鲁木齐830000 [2]新疆医科大学附属中医医院康复科,乌鲁木齐830000 [3]新疆军区总医院全军临床检验诊断中心,乌鲁木齐830000

出　　处：《解放军医药杂志》2020年第10期19-23,共5页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：国家自然科学基金面上项目(81871020)。

摘　　要：目的探讨一氧化氮(NO)诱导星形胶质细胞(ASTs)低氧诱导因子-1α(HIF-1α)活化表达的作用。方法分离新生大鼠大脑ASTs和成年大鼠大脑微血管内皮细胞(MECs),并进行体外原代培养。分别给予ASTs细胞DETA NONOATE预处理和与MECs共培养,观测ASTs细胞HIF-1α活化表达水平的变化。结果常氧条件下,ASTs给予0.8、1.0 mmol/L DETA NONOATE预处理12 h后,HIF-1α蛋白水平高于未给药组(P<0.01);1.0 mmol/L DETA NONOATE给药预处理12、24 h后,HIF-1α蛋白水平高于未给药组(P<0.01)。常氧和低氧组MECs细胞NO释放水平高于ASTs,且MECs在低氧条件下高于常氧组(P<0.01)。常氧条件下,MECs给予内皮型一氧化氮合酶(eNOS)抑制剂L-NAME(100μmol/L)预处理后,NO释放水平降低(P<0.01)。MECs和ASTs共培养后,共培养组ASTs细胞HIF-1α蛋白水平较ASTs单独培养组增高(P<0.01);共培养组给予L-NAME(100μmol/L)预处理后,ASTs中HIF-1α蛋白水平与ASTs单独培养组比较差异无统计学意义(P>0.05)。结论本研究初步观测到体外培养的ASTs在给予2种来源NO后均可诱导HIF-1α活化。Objective To investigate the role of nitric oxide(NO)in inducing activation of hypoxia-inducible factor 1α(HIF-1α)in astrocytes(ASTs).Methods ASTs from neonatal rat brain and microvascular endothelial cells(MECs)from adult rat brain were isolated and cultured in vitro.ASTs were pretreated with DETA NONOATE and co-cultured with MECs to observe changes in the expression of HIF-1αactivation in ASTs.Results Under normoxic conditions,HIF-1αlevels in ASTs treated with 0.8 and 1.0 mmol/L DETA NONOATE for 12 h was higher than that of the non-administration group(P<0.01).At 12 and 24 h after treatment with 1.0 mmol/L DETA NONOATE,HIF-1αlevels were higher than those of the non-administration group(P<0.01).The NO release level of MECs in normoxic and hypoxic groups was higher than that of ASTs(P<0.01),and MECs in hypoxia condition were higher than those in normoxic group(P<0.01).Under normoxic conditions,after MECs were treated with endothelial nitric oxide synthase(eNOS)inhibitor L-NAME(100μmol/L),the NO release level was decreased(P<0.01).After co-culture of MECs and ASTs,the level of HIF-1αin ASTs of the co-culture group was higher than that in the ASTs alone group(P<0.01).After co-culture group was given L-NAME(100μmol/L)treatment,there was no significant difference in the level of HIF-1αin ASTs of the co-culture group compared with that in the ASTs alone group(P>0.05).Conclusion In this study,we observed that in vitro cultured ASTs could induce HIF-1αactivation after treatment with NO from two sources.

关 键 词：一氧化氮 低氧诱导因子-1Α 星形胶质细胞 微血管内皮细胞 原代细胞培养 大鼠 Wistar 

分 类 号：R-332[医药卫生]