中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2012年
5期
286-289
,共4页
姚娜%王迪芬%宋曦%刘鲜林
姚娜%王迪芬%宋晞%劉鮮林
요나%왕적분%송희%류선림
依达拉奉%异丙酚%预处理%脑皮质细胞%缺血/再灌注损伤,脑%谷氨酸%凋亡
依達拉奉%異丙酚%預處理%腦皮質細胞%缺血/再灌註損傷,腦%穀氨痠%凋亡
의체랍봉%이병분%예처리%뇌피질세포%결혈/재관주손상,뇌%곡안산%조망
Edaravone%Propofol%Pretreatment%Cortical neuron%Cereberal ischemia/reperfusion injury%Glutamate%Apoptosis
目的 观察依达拉奉联合异丙酚预处理对乳鼠离体脑皮质细胞缺血/再灌注(VR)损伤的保护作用以及脑保护效应的时间窗.方法 体外培养出生24 h内SD乳鼠脑皮质细胞7d,按随机数字表法分为空白对照组、浴氨酸损伤组、药物预处理24 h对照组及药物预处理24、2、0h组.各药物预处理组分别于谷氨酸损伤(200 μmol/L 0.5 h)前24、2、0h用含100 μmol/L依达拉奉和3 mg/L异丙酚的培养基联合预处理原代培养的神经细胞.通过测定神经细胞存活率[四甲基偶氮唑盐(MTT)比色法]、乳酸脱氢酶(LDH)漏出率、神经细胞Na+-K+-ATP酶活性观察神经细胞成活和细胞损伤情况；通过测定超氧化物歧化酶(SOD)活性(黄嘌呤氧化酶法)、丙二醛(MDA)含量(硫代巴比妥酸法)观察神经细胞抗氧化和氧化水平；流式细胞仪检测神经细胞凋亡情况.结果 与空白对照组比较,谷氨酸损伤组神经细胞存活率[(62.2±23.4)％比(90.5±14.8)％]、SOD活性(U/ml:6.864±2.872比29.569±3.684)、Na+-K+-ATP酶活性(U·mg4·h-1:0.318±0.146比0.636±0.168)均显著下降,神经细胞凋亡率[(9.4±0.7)％比(6.1±0.2)％]、MDA含量(nmol/ml:0.515±0.101比0.294±0.105)、LDH漏出率[(41.2±1.6)％比(36.8±4.6)％]均显著升高(P＜0.05或P＜0.01).与谷氨酸损伤组比较,药物预处理组细胞存活率、SOD活性、Na+-K+-ATP酶活性均显著升高,细胞凋亡率、MDA含量、LDH漏出率均显著下降,并呈时间依赖性,以预处理24 h组作用最为显著[细胞存活率:(89.2±30.3)％比(62.2±23.4)％,SOD活性(U/ml):17.780±4.514比6.864±2.872,Na+-K+-ATP酶活性(U·mg-1·h-1):0.541±0.052比0.318±0.146,细胞凋亡率:(6.7±0.4)％比(9.4±0.7)％,MDA含量(nmol/ml):0.319±0.101比0.515±0.101,LDH漏出率:(37.2±1.4)％比(41.2±1.6)％,均P＜0.01].结论 依达拉奉联合异丙酚预处理对乳鼠离体脑皮质细胞I/R损伤有明显协同保护作用；且以24 h作为时间窗的脑保护作用最明显.
目的 觀察依達拉奉聯閤異丙酚預處理對乳鼠離體腦皮質細胞缺血/再灌註(VR)損傷的保護作用以及腦保護效應的時間窗.方法 體外培養齣生24 h內SD乳鼠腦皮質細胞7d,按隨機數字錶法分為空白對照組、浴氨痠損傷組、藥物預處理24 h對照組及藥物預處理24、2、0h組.各藥物預處理組分彆于穀氨痠損傷(200 μmol/L 0.5 h)前24、2、0h用含100 μmol/L依達拉奉和3 mg/L異丙酚的培養基聯閤預處理原代培養的神經細胞.通過測定神經細胞存活率[四甲基偶氮唑鹽(MTT)比色法]、乳痠脫氫酶(LDH)漏齣率、神經細胞Na+-K+-ATP酶活性觀察神經細胞成活和細胞損傷情況；通過測定超氧化物歧化酶(SOD)活性(黃嘌呤氧化酶法)、丙二醛(MDA)含量(硫代巴比妥痠法)觀察神經細胞抗氧化和氧化水平；流式細胞儀檢測神經細胞凋亡情況.結果 與空白對照組比較,穀氨痠損傷組神經細胞存活率[(62.2±23.4)％比(90.5±14.8)％]、SOD活性(U/ml:6.864±2.872比29.569±3.684)、Na+-K+-ATP酶活性(U·mg4·h-1:0.318±0.146比0.636±0.168)均顯著下降,神經細胞凋亡率[(9.4±0.7)％比(6.1±0.2)％]、MDA含量(nmol/ml:0.515±0.101比0.294±0.105)、LDH漏齣率[(41.2±1.6)％比(36.8±4.6)％]均顯著升高(P＜0.05或P＜0.01).與穀氨痠損傷組比較,藥物預處理組細胞存活率、SOD活性、Na+-K+-ATP酶活性均顯著升高,細胞凋亡率、MDA含量、LDH漏齣率均顯著下降,併呈時間依賴性,以預處理24 h組作用最為顯著[細胞存活率:(89.2±30.3)％比(62.2±23.4)％,SOD活性(U/ml):17.780±4.514比6.864±2.872,Na+-K+-ATP酶活性(U·mg-1·h-1):0.541±0.052比0.318±0.146,細胞凋亡率:(6.7±0.4)％比(9.4±0.7)％,MDA含量(nmol/ml):0.319±0.101比0.515±0.101,LDH漏齣率:(37.2±1.4)％比(41.2±1.6)％,均P＜0.01].結論 依達拉奉聯閤異丙酚預處理對乳鼠離體腦皮質細胞I/R損傷有明顯協同保護作用；且以24 h作為時間窗的腦保護作用最明顯.
목적 관찰의체랍봉연합이병분예처리대유서리체뇌피질세포결혈/재관주(VR)손상적보호작용이급뇌보호효응적시간창.방법 체외배양출생24 h내SD유서뇌피질세포7d,안수궤수자표법분위공백대조조、욕안산손상조、약물예처리24 h대조조급약물예처리24、2、0h조.각약물예처리조분별우곡안산손상(200 μmol/L 0.5 h)전24、2、0h용함100 μmol/L의체랍봉화3 mg/L이병분적배양기연합예처리원대배양적신경세포.통과측정신경세포존활솔[사갑기우담서염(MTT)비색법]、유산탈경매(LDH)루출솔、신경세포Na+-K+-ATP매활성관찰신경세포성활화세포손상정황；통과측정초양화물기화매(SOD)활성(황표령양화매법)、병이철(MDA)함량(류대파비타산법)관찰신경세포항양화화양화수평；류식세포의검측신경세포조망정황.결과 여공백대조조비교,곡안산손상조신경세포존활솔[(62.2±23.4)％비(90.5±14.8)％]、SOD활성(U/ml:6.864±2.872비29.569±3.684)、Na+-K+-ATP매활성(U·mg4·h-1:0.318±0.146비0.636±0.168)균현저하강,신경세포조망솔[(9.4±0.7)％비(6.1±0.2)％]、MDA함량(nmol/ml:0.515±0.101비0.294±0.105)、LDH루출솔[(41.2±1.6)％비(36.8±4.6)％]균현저승고(P＜0.05혹P＜0.01).여곡안산손상조비교,약물예처리조세포존활솔、SOD활성、Na+-K+-ATP매활성균현저승고,세포조망솔、MDA함량、LDH루출솔균현저하강,병정시간의뢰성,이예처리24 h조작용최위현저[세포존활솔:(89.2±30.3)％비(62.2±23.4)％,SOD활성(U/ml):17.780±4.514비6.864±2.872,Na+-K+-ATP매활성(U·mg-1·h-1):0.541±0.052비0.318±0.146,세포조망솔:(6.7±0.4)％비(9.4±0.7)％,MDA함량(nmol/ml):0.319±0.101비0.515±0.101,LDH루출솔:(37.2±1.4)％비(41.2±1.6)％,균P＜0.01].결론 의체랍봉연합이병분예처리대유서리체뇌피질세포I/R손상유명현협동보호작용；차이24 h작위시간창적뇌보호작용최명현.
Objective To investigate the protective effect of combined pretreatment of edaravone and propofol on cerebral cortex with ischemia/reperfusion ( I/R ) injury and its therapeutic window.Methods Sprague Dawley (SD) rat brain cortex cells harvested within 24 hours of birth were cultured in vitro for 7 days.The cells were then divided into blank control group,glutamate injury group,24-hour drug precondition control group,and 24-,2-,0-hour drug precondition groups according to random number table.The nerve cells in each pretreatment group were cultured in medium containing 100 μmol/L of edaravone and 3 mg/L of propofol 24,2,or 0 hour before glutamate damage (200 μmol/L for 0.5 hour).Nerve cell survival or damage was determined by methyl thiazolyl tetrazolium (MTT),lactate dehydrogenase (LDH) leakage rate,and nerve cell Na+-K+-ATPase activity.The oxidation and anti-oxidation ability of nerve cells was observed by determining superoxide dismutase (SOD) activity (xanthine oxidase),malondialdehyde (MDA) content (thiobarbituric acid).Nerve apoptosis was detected by flow cytometry.Results Compared with blank control group,in the glutamate injury group,nerve cell survival rate [ (62.2 ± 23.4)％ vs.(90.5 ± 14.8)％],the activity of SOD (U/ml:6.864 ± 2.872 vs.29.569 ± 3.684),Na+-K+-ATPase activity ( U· mg-1· h-1:0.318 ± 0.146 vs.0.636 ± 0.168 ) were significantly decreased,and rate of neuronal apoptosis [ (9.4 ±0.7)％ vs. (6.1 ±0.2)％ ],the content of MDA (nmol/ml:0.515 ±0.101 vs.0.294 ±0.105),LDH leakage rate [(41.2 ± 1.6)％ vs.(36.8 ±4.6)％]were significantly increased (P＜0.05 or P＜0.01 ).Compared with glutamate injury group,the cell survival rate and the activity of SOD and Na+-K+-ATPase were significantly increased in the drug pretreatment groups,and apoptosis rate,MDA content,and LDH leakage rate were significantly decreased with time-department,and effect in the 24-hour pretreatment group was most significant [survival rate of cell: (89.2 ±30.3)％vs. (62.2±23.4)％,SOD activity (U/ml):17.780±4.514 vs.6.864±2.872,Na+-K+-ATPase activity ( U· mg-1· h-1 ):0.541 ± 0.052 vs.0.318 ± 0.146,the rate of cell apoptosis:( 6.7 ± 0.4 )％ vs.(9.4 ± 0.7 ) ％,the content of MDA (nmol/ml):0.319±0.101 vs.0.515±0.101,LDHleakagerate: (37.2±1.4)％vs.(41.2±1.6)％,all P＜0.01].Conclusion The synergistic protective effect of pretreatment with edaravone combined with propofol on neonatal rat brain cortex cells with I/R injury in vitro was evident; and 24-hour pretreatment is the best time window of protection for the cerebral neurons.
