文章摘要
不同浓度右美托咪定对糖氧剥夺/再灌注诱导神经细胞凋亡的影响
Effects of dexmedetomidine on oxygen-glucose deprivation/reperfusion-induced neuronal apoptosis
  
DOI：
中文关键词: 糖氧剥夺/再灌注  内质网应激  中脑星形胶质细胞源性神经营养因子  右美托咪定  神经保护
英文关键词: Oxygen-glucose deprivation/reperfusion  ER stress  Mesencephalic astrocyte-derived neurotrophic factor  Dexmedetomidine  Neuroprotection
基金项目:安徽省2015年第一批科技计划项目对外科技合作计划（1503062021）
作者单位E-mail
康恺 230000,合肥市,安徽医科大学附属省立医院麻醉科  
康芳 230000,合肥市,安徽医科大学附属省立医院麻醉科  
沈玉君 230000,合肥市,安徽医科大学附属省立医院麻醉科  
沈玉先 230000,合肥市,安徽医科大学附属省立医院麻醉科  
黄祥 230000,合肥市,安徽医科大学附属省立医院麻醉科  
李娟 230000,合肥市,安徽医科大学附属省立医院麻醉科 huamuzi1999@126.com 
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中文摘要:
      
目的 研究不同浓度右美托咪定对糖氧剥夺/再灌注(OGD/R)诱导神经细胞凋亡的保护作用。
方法 应用全反式维甲酸(ATRA)和十四烷酰佛波醇乙酯酸(TPA)序贯诱导人源性神经母细胞瘤(SH-SY5Y)分化为神经细胞,均分为六组。选择OGD12 h/R12 h构建OGD/R模型。D0、D1、D2、D3、D4、D5组在OGD开始即刻分别加入右美托咪定0、0.1、1、10、100、1 000 μmol/L。于再灌注12 h后,采用MTT法检测细胞存活率,流式细胞凋亡法检测细胞凋亡水平,以观察不同浓度右美托咪定对OGD/R诱导神经细胞凋亡的保护作用。随后选择保护效果确切组,应用Western-blot法检测内质网(endoplasmic reticulum, ER)应激特异性蛋白-中脑星形胶质细胞源性神经营养因子(mesencephalic astrocyte-derived neurotrophic factor,MANF)含量以及促凋亡蛋白Caspase-3活性和CHOP含量。
结果 与D0组比较,D1、D2组细胞存活率和细胞凋亡率差异无统计学意义;D4、D5组细胞存活率明显下降,细胞凋亡率明显升高(P<0.01或P<0.05);D3组则显著改善并提高了OGD/R诱导后神经细胞的存活率,抑制了细胞凋亡(P<0.01)。Western blot结果显示,D3组细胞MANF含量明显高于D0组(P<0.01),Caspase-3活性和CHOP含量明显低于D0组(P<0.01)。
结论 右美托咪定在10 μmol/L终浓度时对OGD/R诱导的神经细胞凋亡具有保护作用,并显著提高了细胞存活率。右美托咪定神经保护作用的机制可能与上调ER应激特异性蛋白MANF,抑制凋亡蛋白Caspase-3和CHOP的表达相关。
英文摘要:
      
Objective To investigate protective effects of dexmedetomidine on oxygen-glucose deprivation/reperfusion (OGD/R)-induced neuronal apoptosis.
Methods SH-SY5Y cells were differentiated to neurons with ATRA and followed by TPA. According to the results of preliminary experiment, OGD/R modle was constructed by oxygen-glucose deprivation(OGD) for 12 h and reperfusion (R) for another 12 h. During the start of the OGD, neurons were immediately divided into six groups: group D0 (0 μmol/L dexmedetomidine), group D1 (0.1 μmol/L dexmedetomidine), group D2 (1 μmol/L dexmedetomidine), group D3 (10 μmol/L dexmedetomidine), group D4 (100 μmol/L dexmedetomidine), group D5 (1 000 μmol/L dexmedetomidine). After reperfusion 12 h, the cell viability was evaluated by the method of MTT. The cellular apoptosis was observed by flow cytometry method. The protective effects of different concentration dexmedetomidine on OGD/R-induced neuronal apoptosis were investigated. Then in chosen the exact group having protective effects, endoplasmic reticulum stress specific protein mesencephalic astrocyte derived neurotrophic factor (MANF) and pro-apoptotic protein Caspase-3 and CHOP were detected by Western blot method.
Results Compared with group D0, there was no difference on the cell viability and cellular apoptosis induced by OGD/R in groups D1 and D2, but a significant decrease and increase in groups D4 and D5 (P<0.01 or P<0.05). And only group D3 had a neuroprotective effect, significantly increased the cell viability and inhibited the apoptosis (P<0.01). Further studys found that group D3 significantly up-regulated ER stress specific protein MANF (P<0.01) and inhibited up-regulation of Caspase-3 and CHOP (P<0.01).
Conclusion These data suggest that 10 μmol/L dexmedetomidine had neuroprotective effect on OGD/R-induced neuronal apoptosis and significantly increased cell viability. Our results also indicate that up-regulation of ER stress specific protein MANF and inhibition of CHOP and Caspase-3 by MANF are involved in the neuroprotective effects of dexmedetomidine.
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