摘要
目的评价七氟醚预处理联合后处理对新生大鼠心肌细胞缺氧复氧损伤的影响。方法健康新生SD大鼠，1～3d龄，处死后取心室肌组织，分离心肌细胞，将心肌细胞于DMEM培养液中培养，细胞密度3×105个／ml，接种于24孔培养板（1ml/孔）、35mm培养皿（5ml／皿）或50ml培养瓶（8ml／瓶），采用随机数字表法，将其随机分为9组（n=24）：正常对照组（C组）常规培养160min；缺氧复氧组（A／R组）、七氟醚预处理组（S1组）、七氟醚预处理＋SB203580组（S1＋SB组）、七氟醚后处理组（S2组）、七氟醚后处理＋SB203580组（S2＋SB组）、七氟醚预处理＋七氟醚后处理组（S3组）、七氟醚预处理＋七氟醚后处理＋SB203580组（S3＋SB组）和SB203580组（SB组）均进行缺氧120min，复氧20min。S1组、S1＋SB组、S3组和S3＋SB组缺氧前用2．5％七氟醚孵育20min，S1＋SB组和S3＋SB组七氟醚预处理同时加入5μmol／Lp38丝裂原激活蛋白激酶（p38MAPK）特异性抑制剂SB203580；S2组、S2＋sB组、s3组和s，＋sB组复氧开始时采用2．5％七氟醚孵育20min，S2＋SB组和S3＋SB组于七氟醚后处理同时加入5μmol／LSB203580；SB组缺氧前20min和复氧开始时均用5μmol／LSB203580孵育20min。复氧结束时测定细胞培养液乳酸脱氢酶活性、细胞存活率和细胞凋亡率，分别于七氟醚预处理结束和后处理结束时测定磷酸化p38MAPK（p-p38MAPK）表达水平。结果与C组比较，其余各组乳酸脱氢酶活性和细胞凋亡率升高，细胞存活率降低（P〈0．05）；与A／R组比较，S1组、S2组和S3组LDH活性和细胞凋亡率降低，细胞存活率升高（P〈0．05）；S1组、S2组和S3组间上述指标差异无统计学意义（P〉0．05）；SB203580可取消七氟醚预处理、后处理及二者联合应用时心肌保护效应（P〈0．05）。七氟醚预处理和后处理均可上调p-p38MAPK表达，且二者联合应用时上调p
Objective To investigate the effect of sevoflurane preconditioning combined with postcondi- tioning （Spost） on anoxia/reoxygenation （A/R） injury to neonatal rat cardiomyocytcs. Methods Primary cultured neonatal rat eardiomyocytes were isolated from SD rats aged 1-3 days and cultured in DMEM liquid culture medi- um. The cells were seeded in 24-well plates （ 1 ml/hole）, 35 mm diameter dishes （5 ml/dish） or in 50 mm culture flasks （8 ml/flask） with a density of 3 × 105/rnl and randomly divided into 9 groups （ n = 24 each） ： control group （group C）, A/R group, Spre group （group S1 ）, Spre ＋ SB203580 group （group S1 ＋ SB）, sevoflurane postcon- ditioning （Spost） group （group S2 ）, Spost ＋ SB203580 group （ group S2 ＋ SB）, Spre ＋ Spost group （ group S3 ）,Spre ＋ Spost ＋ SB203580 group （group S3 ＋ SB）, and group SB203580 （group SB）. The cells were cultured rou- tinely for 160 min in group C and the cells were exposed to 95 % N2-5 % CO2 in an incubator at 37 ℃ for 120 min followed by reoxygenation for 20 rain in the other groups. The cells were incubated with 2,5 % sevoflurane for 20 min before anoxia in groups Sl , S1 ＋ SB, S3 and S3 ＋ SB and in addition SB203580 （specific p38MAPK inhibitor） 5 μmol/L was added simultaneously in groups S1 ＋ SB and S3 ＋ SB. The cells were incubated with 2.5% sevoflu- rane for 20 rain after beginning of reoxygenation in groups S2 , S2 ＋ SB, S3 and S3 ＋ SB, and in addition SB203580 5 μmol/L was added simultaneously in groups S2 ＋ SB andS3 ＋ SB. The cells were incubated with SB203580 5 μmol/L for 20 rain before anoxia and after beginning of reoxygenation in group SB. The lactate dehydrogenase （LDH） activity, cell survival rate and apoptotic rate were measured at the end of reoxygenation. The levels of phosphor-p38MAPK （p-p38MAPK） was detected at the end of Spre and Spost. Results Compared with group C, the LDH activity and apoptotic rate were significantly increased, while the cell su
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2012年第8期1013-1016,共4页
Chinese Journal of Anesthesiology
关键词
麻醉药
吸入
细胞低氧
肌细胞
心脏
缺血预处理
婴儿
新生
后处理
Anesthetics, inhalation
Cell hypoxia
Myocytes, cardiac
Ischemic preconditioning
Infant, newborn
Postconditioning
