右美托咪定对氧糖剥夺后星形胶质细胞反应性增殖的影响  

Effects of Dexmedetomidine on Reactive Astrocyte Proliferation after Oxygen-glucose Deprivation

作　　者：廖明锋[1] 迟晓慧[1] 杨柳[1] 赵以林[1] 冯丽[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院麻醉学教研室,武汉430030

出　　处：《华中科技大学学报（医学版）》2016年第3期258-262,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81500982);湖北省卫生计生委资助项目(No.WJ2015MB008)

摘　　要：目的探讨右美托咪定对氧糖剥夺后星形胶质细胞反应性增殖的影响。方法采用出生1~2d的SD大鼠,提取原代星形胶质细胞,将其随机分为5组：对照组（C组）、氧糖剥夺/复氧（OGD/R）组、右美托咪定＋OGD/R组（Dex＋OGD/R组）、α2肾上腺能受体抑制剂育亨宾＋OGD/R组（YHB＋OGD/R组）和右美托咪定＋育亨宾＋OGD/R组（Dex＋YHB＋OGD/R组）。缺糖缺氧1h后,复糖复氧培养24h制备OGD/R模型。C组星形胶质细胞仅做正常培养;OGD/R组星形胶质细胞制备OGD/R模型;Dex＋OGD/R组星形胶质细胞于OGD处理1h后,于复糖的培养液中加入终浓度为10μmol/L的右美托咪定;YHB＋OGD/R组星形胶质细胞于OGD/R处理前30min向培养液中加入终浓度为10μmol/L的育亨宾,之后制备OGD/R模型;Dex＋YHB＋OGD/R组于OGD/R处理前30min向培养液中加入终浓度为10μmol/L的育亨宾,其它处理同Dex＋OGD/R组。EdU法检测细胞增殖能力;Western blot法测定增殖细胞核抗原（PCNA）、胶质纤维酸性蛋白（GFAP）、磷酸化STAT3（p-STAT3）的蛋白表达水平;ELISA法测定培养液中白细胞介素-6（IL-6）和睫状神经营养因子（CNTF）的浓度。结果与C组比较,OGD/R组EdU阳性细胞率,PCNA、GFAP、pSTAT蛋白表达水平,培养液中IL-6和CNTF释放量均显著增高（均P〈0.05）;与OGD/R组比较,Dex＋OGD/R组EdU阳性细胞率,PCNA、GFAP、p-STAT蛋白表达水平,培养液中IL-6和CNTF释放量较低（均P〈0.05）;与Dex＋OGD/R组比较,Dex＋YHB＋OGD/R组EdU阳性细胞率,PCNA、GFAP、p-STAT3蛋白表达水平,培养液中IL-6和CNTF释放量均较高（均P〈0.05）。结论右美托咪定可缓解氧糖剥夺后星形胶质细胞反应性增殖。Objective To investigate the effects of dexmedetomidine on reactive astrocyte proliferation after oxygen-glucose deprivation.Methods Primary astrocytes were harvested from newborn SD rats of postnatal day 1-2.They were randomly di-vided into 5 groups：control group （Group C）,oxygen-glucose deprivation/reperfusion （OGD/R）group （Group OGD/R）, dexmedetomidine＋OGD/R group （Group Dex＋OGD/R）,yohimbine＋OGD/R group （Group YHB＋OGD/R）and dexme-detomidine＋yohimbine＋OGD/R group （Group Dex＋YHB＋OGD/R）.The model of OGD/R was established by replacing the standard astrocyte culture medium （DMEM/F12）with glucose-free medium and then immediately exposing cells to 95%N2 and 5% CO2 in the anaerobic chamber at 37＆#176;C for 1 h,followed by a return to standard culture conditions for another 24 h.The astrocytes in Group OGD/R were subj ected to OGD/R treatment as described above.Those in Group C were not ex-posed to OGD.The treatment for Group Dex＋OGD/R was the same as for Group OGD/R,except that 10μmol/L dexmedeto-midine was added to the normal medium during reoxygenation.The treatment for Group YHB＋OGD/R was the same as for Group OGD/R,except that 10 μmol/L ofα2 adrenergic receptor antagonist yohimbine was added to medium 30 min before OGD/R treatment.In Group Dex＋YHB＋OGD/R,10μmol/L yohimbine was added to medium 30 min before OGD/R treat-ment and 10μmol/L dexmedetomidine to medium during reoxygenation.The 5-ethynyl-2’-deoxyuridine （EdU）assay was used to evaluate the astrocyte proliferation after OGD/R treatment.Western blotting was applied to detect the protein levels of pro-liferating cell nuclear antigen （PCNA）and glial fibrillary acidic protein （GFAP）,phospho-signal transducer and activator of transcription 3 （STAT3）.ELISA assay was used to determine the concentrations of interlukin-6 （IL-6）and ciliary neurotrophic factor （CNTF）in the medium.Results The proportion of EdU-positive cells,the protein levels of PCNA,GFAP,p-STAT,and the release of IL-

关 键 词：右美托咪定 氧糖剥夺 胶质细胞反应性增殖 缺血性损伤 

分 类 号：R971.3[医药卫生—药品]