CAJ | 학술논문

目的观察马来酸桂哌齐特预处理对大鼠脑缺血半暗带区胞外信号调节激酶(ERK1/2)磷酸化表达的影响，探讨其对脑缺血的保护作用及可能的作用机制。方法采用改良线栓法建立大鼠永久性大脑中动脉阻塞(pMCAO)模型。SD大鼠随机分为①马来酸桂哌齐特组(pMCAO模型大鼠，n=57。尾静脉注射马来酸桂哌齐特3.0 mg·kg-1，×5 d)；②对照组(pMCAO模型大鼠，n=57。尾静脉注射生理盐水0.5 mL，×5 d)；③假手术组(不插线栓，n=50。尾静脉注射生理盐水0.5 mL，×5 d)；④正常组(n=3，不做任何处理)。应用TTC染色法测定梗死体积，蛋白印迹法和免疫组化法检测不同时间点(术后3、6、24、48和72 h)缺血半暗带区ERK蛋白的表达。结果马来酸桂哌齐特组与对照组比较，梗死体积减少17.91%(P=0.001)。马来酸桂哌齐特组缺血半暗带pERK1/2表达于3 h开始增加，24 h达高峰[为正常的(5.75±0.70)倍]后逐渐降低，72 h仍为正常的(2.89±0.51)倍，各时间点与正常组比较，均差异有显著统计学意义(P<0.01)。6、24和48 h 3个时间点，马来酸桂哌齐特组缺血半暗带内pERK1/2表达较对照组增加(P<0.05)。pERK的免疫组化检测示马来酸桂哌齐特预处理能上调缺血半暗带区内ERK磷酸化表达(P<0.05)。结论马来酸桂哌齐特预处理可减少脑梗死体积，并能上调缺血半暗带区的ERK磷酸化表达；参与MAPK信号通路、上调缺血半暗带区内ERK磷酸化的表达，可能是其保护缺血性脑损伤的机制之一。
목적관찰마래산계고제특예처리대대서뇌결혈반암대구포외신호조절격매(ERK1/2)린산화표체적영향，탐토기대뇌결혈적보호작용급가능적작용궤제。방법채용개량선전법건립대서영구성대뇌중동맥조새(pMCAO)모형。SD대서수궤분위①마래산계고제특조(pMCAO모형대서，n=57。미정맥주사마래산계고제특3.0 mg·kg-1，×5 d)；②대조조(pMCAO모형대서，n=57。미정맥주사생리염수0.5 mL，×5 d)；③가수술조(불삽선전，n=50。미정맥주사생리염수0.5 mL，×5 d)；④정상조(n=3，불주임하처리)。응용TTC염색법측정경사체적，단백인적법화면역조화법검측불동시간점(술후3、6、24、48화72 h)결혈반암대구ERK단백적표체。결과마래산계고제특조여대조조비교，경사체적감소17.91%(P=0.001)。마래산계고제특조결혈반암대pERK1/2표체우3 h개시증가，24 h체고봉[위정상적(5.75±0.70)배]후축점강저，72 h잉위정상적(2.89±0.51)배，각시간점여정상조비교，균차이유현저통계학의의(P<0.01)。6、24화48 h 3개시간점，마래산계고제특조결혈반암대내pERK1/2표체교대조조증가(P<0.05)。pERK적면역조화검측시마래산계고제특예처리능상조결혈반암대구내ERK린산화표체(P<0.05)。결론마래산계고제특예처리가감소뇌경사체적，병능상조결혈반암대구적ERK린산화표체；삼여MAPK신호통로、상조결혈반암대구내ERK린산화적표체，가능시기보호결혈성뇌손상적궤제지일。
Aim To observe the influence of cinepazide maleate pretreatment on the expression of extracellular signal-regulated kinase (ERK1/2) phosphorylation in cerebral ischemic penumbra in the permanent middle cerebral artery occlusion (pMCAO) model rats, and explore its neuroprotective function and other possible mechanism.MethodsThe pMCAO model rats were established by modified suture embolization method. SD rats were randomly divided into a cinepazide maleate group (n=57) (pMCAO model rats, tail intravenous injection of cinepazide maleate 3.0 mg·kg-1 for 5 days), a control group (n=57) (pMCAO model rats, tail intravenous injection of normal saline 0.5 mL·d-1 for 5 days) and a sham group (n=50) (the same model of pMCAO but no suture inserted, tail intravenous injection of normal saline 0.5 mL·d-1 for 5 days). Infarct volume was determined by TTC staining, the expression of ERK1/2 phosphorylation in cerebral ischemic penumbra at different time points after cerebral ischemia (3, 6, 24, 48 and 72h) were detected by Western blotting and immunohistochemistry.ResultsCompared with the control group, the infarct volum of cinepazide maleate groupe reduced by 17.91% (P=0.001 ). The expression of pERK1/2 in ischemic penumbra of cinepazide maleate group began to increase at 3 hours of ischemia, and reached the peak at 24 hours [(5.75±0.70) times of the normal]. Then it was still (2.89±0.51) times of the normal at 72 hours of ischemia. Compared with the normal group, each time point have signiifcant statistical differences (P<0.01). At the three time points (6 h, 24 h and 48 h), the expression of pERK1/2 of cinepazide maleate group compared with that of the control group increased deifnitely in the ischemic penumbra (P<0.05). Through immunohistochemistry staining, it indicated the cinepazide maleate pretreatment could up-regulate the expression of phosphorylated ERK in ischemic penumbra (P<0.05). Conclusion Cinepazide maleate pretreatment could reduce infarct volume and increase the expression of ERK phosphorylation in ischemic penumbra. With participating in the MAPK signaling pathway, the increase of the expression of ERK phosphorylation in ischemic penumbra may be one of the mechanisms for the protection of ischemic brain injury.