中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2010年
4期
226-229,前插2
,共5页
神经生长因子%依达拉奉%谷氨酸%预处理%缺血/再灌注损伤,脑
神經生長因子%依達拉奉%穀氨痠%預處理%缺血/再灌註損傷,腦
신경생장인자%의체랍봉%곡안산%예처리%결혈/재관주손상,뇌
Nerve growth factor%Edaravone%Glutamate%Pretreatment%Cerebral ischemia/ reperfusion injury
目的 比较神经生长因子(NGF)和自由基清除剂依达拉奉预处理对脑缺血/再灌注(I/R)损伤神经细胞的保护作用.方法 取出生24 h内的SD乳鼠脑皮质细胞,体外培养7 d后分为对照组,药物损伤组,NGF 10、50、100 μg/L预处理组及依达拉奉100 μmol/L预处理组.各预处理组分别预处理24 h后,给予200μmol/L谷氨酸(Glu)损伤0.5 h,换正常培养液继续培养24 h;培养9 d时检测神经细胞存活率[四甲基偶氮唑盐(MTT)比色法]、乳酸脱氢酶(LDH)漏出率(分光光度法)、细胞凋亡率(流式细胞术);用苏木素-伊红(HE)染色后在倒置相差显微镜下观察细胞形态变化,在透射电镜下观察细胞超微结构改变.结果 与药物损伤组比较,NGF 10、50、100μg/L预处理组和依达拉奉预处理组细胞存活率明显升高C(0.21±0.04)%、(0.23±0.04)%、(0.21±0.04)%、(0.24±0.04)%比(0.19±0.04)%],LDH漏出率[(0.50±0.06)%、(0.46±0.07)%、(0.50±0.02)%,(0.43±0.06)%比(0.56±0.03)%]和细胞凋亡率[(10.77±1.07)%、(10.38±0.70)%、(13.34±0.57)%、(9.99±0.77)%比(14.52±0.77)%]均不同程度降低(P<0.05或P<0.01);细胞形态受损程度和神经元超微结构病变均减轻.NGF 3个浓度组中以50μg/L组效果最佳,且与依达拉奉组各指标差异无统计学意义.结论 NGF和依达拉奉预处理脑皮质细胞24 h对脑I/R损伤均有保护作用;50 μg/L NGF和100 μmol/L依达拉奉预处理效果最佳,且二者最佳浓度时疗效相当.
目的 比較神經生長因子(NGF)和自由基清除劑依達拉奉預處理對腦缺血/再灌註(I/R)損傷神經細胞的保護作用.方法 取齣生24 h內的SD乳鼠腦皮質細胞,體外培養7 d後分為對照組,藥物損傷組,NGF 10、50、100 μg/L預處理組及依達拉奉100 μmol/L預處理組.各預處理組分彆預處理24 h後,給予200μmol/L穀氨痠(Glu)損傷0.5 h,換正常培養液繼續培養24 h;培養9 d時檢測神經細胞存活率[四甲基偶氮唑鹽(MTT)比色法]、乳痠脫氫酶(LDH)漏齣率(分光光度法)、細胞凋亡率(流式細胞術);用囌木素-伊紅(HE)染色後在倒置相差顯微鏡下觀察細胞形態變化,在透射電鏡下觀察細胞超微結構改變.結果 與藥物損傷組比較,NGF 10、50、100μg/L預處理組和依達拉奉預處理組細胞存活率明顯升高C(0.21±0.04)%、(0.23±0.04)%、(0.21±0.04)%、(0.24±0.04)%比(0.19±0.04)%],LDH漏齣率[(0.50±0.06)%、(0.46±0.07)%、(0.50±0.02)%,(0.43±0.06)%比(0.56±0.03)%]和細胞凋亡率[(10.77±1.07)%、(10.38±0.70)%、(13.34±0.57)%、(9.99±0.77)%比(14.52±0.77)%]均不同程度降低(P<0.05或P<0.01);細胞形態受損程度和神經元超微結構病變均減輕.NGF 3箇濃度組中以50μg/L組效果最佳,且與依達拉奉組各指標差異無統計學意義.結論 NGF和依達拉奉預處理腦皮質細胞24 h對腦I/R損傷均有保護作用;50 μg/L NGF和100 μmol/L依達拉奉預處理效果最佳,且二者最佳濃度時療效相噹.
목적 비교신경생장인자(NGF)화자유기청제제의체랍봉예처리대뇌결혈/재관주(I/R)손상신경세포적보호작용.방법 취출생24 h내적SD유서뇌피질세포,체외배양7 d후분위대조조,약물손상조,NGF 10、50、100 μg/L예처리조급의체랍봉100 μmol/L예처리조.각예처리조분별예처리24 h후,급여200μmol/L곡안산(Glu)손상0.5 h,환정상배양액계속배양24 h;배양9 d시검측신경세포존활솔[사갑기우담서염(MTT)비색법]、유산탈경매(LDH)루출솔(분광광도법)、세포조망솔(류식세포술);용소목소-이홍(HE)염색후재도치상차현미경하관찰세포형태변화,재투사전경하관찰세포초미결구개변.결과 여약물손상조비교,NGF 10、50、100μg/L예처리조화의체랍봉예처리조세포존활솔명현승고C(0.21±0.04)%、(0.23±0.04)%、(0.21±0.04)%、(0.24±0.04)%비(0.19±0.04)%],LDH루출솔[(0.50±0.06)%、(0.46±0.07)%、(0.50±0.02)%,(0.43±0.06)%비(0.56±0.03)%]화세포조망솔[(10.77±1.07)%、(10.38±0.70)%、(13.34±0.57)%、(9.99±0.77)%비(14.52±0.77)%]균불동정도강저(P<0.05혹P<0.01);세포형태수손정도화신경원초미결구병변균감경.NGF 3개농도조중이50μg/L조효과최가,차여의체랍봉조각지표차이무통계학의의.결론 NGF화의체랍봉예처리뇌피질세포24 h대뇌I/R손상균유보호작용;50 μg/L NGF화100 μmol/L의체랍봉예처리효과최가,차이자최가농도시료효상당.
Objective To compare the protective effect of nerve growth factor (NGF) and edaravone (a free radical scavenger) pretreatment against cerebral ischemia/reperfusion (I/R) injury to nerve cells. Methods Cortical neurons of Sprague-Dawley (SD) mouse aged shorter than 24 hours were cultured for 7 days, then they were randomly divided into control group, I/R group, NGF of 10, 50, 100 μg/L pretreatment groups and 100 μmol/L edaravone pretreatment group. In pretreatment groups the cells were pretreated with drugs correspondingly. After culturing for 24 hours, glutamate of 200 μmol/L was given into the culture of all groups, except control group, for half an hour. Then culture medium in all groups were renewed with ordinary culture medium, and cultures were continued for 24 hours. The survival rate (by methyl thiazolyl tetrazolium assay), the content of lactate dehydrogenase (LDH, by spectrometry) and the rate of apoptosis (by flow cytometric) were determined. The cellular shape and ultrastructure were observed by hematoxylin-eosin (HE) staining and electronic microscopy correspondingly. Results The survival rate of nerve cells in NGF groups and edaravone group was significantly higher than that in I/R group [(0.21± 0.04)%, (0.23±0.04)%, (0.21±0.04)%, (0.24±0.04)% vs. (0.19±0.04)%. The content of LDH in culture medium and the rate of apoptosis in NGF groups and edaravone group were lower than those in I/R group (P<0.05 or P<0.01). The release rate of LDH in each group was (0.50±0.06)%, (0.46± 0.07)%, (0.50±0.02)%, (0.43±0.06)% vs. (0.56±0.03)%, respectively. The rate of apoptosis in each group was (10.77±1.07)%, (10.38±0.70)%, (13.34±0.57)%, (9.99±0.77)% vs. (14.52± 0.77)%, respectively. The cellular shape and ultrastructure of nerve cells in NGF groups and edaravone group were affected much less than that of I/R group. NGF of 50 μg/L pretreatment group gave the best effect among three groups. There was no significant difference between NGF 50 μg/L pretreatment group and edaravone pretreatment group. Conclusion NGF and edaravone pretreatment 24 hours before cerebral I/R give protective effects against cerebral I/R injury. The protective effects are best in NGF of 50 μg/L pretreatment group and edaravone pretreatment group, and there is no significant difference between them.
