大鼠脑缺血-再灌注后G蛋白偶联受体84的表达升高  

Increased expression of G protein-coupled receptor 84 after cerebral ischemia-reperfusion in rats

作　　者：赵翀翀[1] 郭文婷 蔡宏斌[1] 王欢[1] ZHAO Chong-chong;GUO Wen-ting;CAI Hong-bin;WANG Huan(Department of Neurology, the Second Hospital of Lanzhou University, Lanzhou 730030, China)

机构地区：[1]兰州大学第二医院神经内科,甘肃兰州730030

出　　处：《基础医学与临床》2021年第3期398-403,共6页Basic and Clinical Medicine

基　　金：甘肃省自然科学基金(20JR5RA330)。

摘　　要：目的探讨G蛋白偶联受体84(GPR84)在大鼠脑缺-再灌注(I/R)后的表达及其功能。方法构建大鼠大脑中动脉阻断(MCAO)模型,将大鼠分为假手术组(sham)和脑缺血2 h后再灌注12 h组(I/R 12 h)、24 h组(I/R 24 h)、48 h组(I/R 48 h)、72 h组(I/R 72 h);体外培养大鼠原代皮质神经元,将细胞分成对照组(control)、氧糖剥夺1 h复氧复糖24 h组(OGD/R 1 h)和氧糖剥夺2 h复氧复糖24 h组(OGD/R 2 h);GPR84小干扰RNA(siRNA)转染神经元,将细胞分为对照组(control)、氧糖剥夺1 h复氧复糖24 h组(OGD/R 1 h)、神经元转染干扰对照后氧糖剥夺1 h复氧复糖24 h组(NC+OGD/R 1 h)和神经元转染GPR84 siRNA后氧糖剥夺1 h复氧复糖24 h组(GPR84 siRNA+OGD/R 1 h);Western blot检测GPR84的表达;免疫荧光检测脑组织中GPR84蛋白的定位;DNA断裂的原位末端标记(TUNEL)检测神经元的凋亡比例。结果与假手术组相比,大鼠脑I/R后各时间点GPR84蛋白水平明显升高(P<0.01)。GPR84主要定位于神经元和小胶质细胞,星形胶质细胞几乎不表达,并且神经元中GPR84的表达在脑I/R后明显提高。与对照组相比,OGD/R 1 h和OGD/R 2 h组神经元中GPR84蛋白的表达均明显提高(P<0.01),并且GPR84干扰可以明显抑制氧糖剥夺后复氧复糖诱导的GPR84表达上调。与对照组相比,OGD/R组神经元凋亡明显提高(P<0.01);与NC+OGD/R 1 h组相比,GPR84 siRNA+OGD/R 1 h组神经元凋亡明显减少。结论脑I/R诱导的GPR84表达升高在脑缺血性损伤中发挥着重要作用。Objective To determine the expression and function of G protein-coupled receptor 84(GPR84)in rat cerebral ischemia-reperfusion(I/R).Methods The rat middle cerebral artery occlusion(MCAO)models were performed,and rats were then divided into sham group,cerebral ischemia for 2 h and reperfusion for 12 h group(I/R 12 h),24 h group(I/R 24 h),48 h group(I/R 48 h),72 h group(I/R 72 h);rat primary cortical neurons were isolated and cultured in vitro,neurons were then divided into control group(control),oxygen-glucose deprivation(OGD)for 1 h and recovery for 24 h group(OGD/R 1 h),OGD for 2 h and recovery for 24 h group(OGD/R 2 h);neurons were transfected with GPR84 small interfering RNA(siRNA),cells were divided into control group,OGD/R 1 h group,neurons pretreated with negative control and exposed to OGD for 1 h and recoveryfor 24 h group(NC+OGD/R 1 h),neurons pretreated with GPR84 siRNA and exposed to OGD for 1 h and recovery for 24 h group(GPR84 siRNA+OGD/R 1 h).Western blot was used to determine the expression of GPR84 protein.Immunofluorescence assay was used to analyze the localization of the immunofluorescence signals of GPR84 in cerebral ischemic tissues after MCAO.TdT-mediated dUTP nick end labeling(TUNEL)assay was used to detect the percentage of apoptotic neurons.Results Compared with the sham group,the expression of GPR84 protein was significantly up-regulated at each time point after cerebral I/R(P<0.01).In addition,GPR84 was mainly localized in neurons and microglia,while little GPR84 was found in astrocytes.Neurons in the MCAO group showed a higher level of GPR84 expression than those in the sham group.GPR84 knockdown significantly inhibited the increase of GPR84 expression induced by OGD/R.Compared with the control group,neurons in the OGD/R 1 h group exhibited higher apoptotic cells percentage.Compared with the NC+OGD/R 1 h group,the percentage of apoptotic neurons in GPR84 siRNA+OGD/R 1 h group was significantly decreased.Conclusions These results suggest that the elevated GPR84 expression induced by cer

关 键 词：GPR84 神经元 脑缺血 凋亡 

分 类 号：R743.3[医药卫生—神经病学与精神病学]