摘要
目的：评价哺乳动物雷帕霉素靶蛋白（mTOR ）在缺氧预处理（HPC ）减轻乳鼠离体心肌细胞氧糖缺失/复氧复糖（OGD/R ）损伤中的作用。方法选择出生1～3 d的SD大鼠，分离培养原代心肌细胞，细胞接种于96孔板（细胞密度1×105/ml ，200μl/孔），采用随机数字表法，将其分为6组（ n＝6）：对照组（C组）常规培养；雷帕霉素组（R组）培养基中加入雷帕霉素（终浓度20 nmol/L ）孵育1 h；OGD/R组；R＋OGD/R组培养基中加入雷帕霉素（终浓度20 nmol/L ）孵育1 h后行OGD/R；HPC＋OGD/R组完成HPC后即刻行OGD/R；R＋HPC＋OGD/R组培养基中加入雷帕霉素（终浓度20 nmol/L ）孵育1h后行HPC及OGD/R。采用氧糖缺失2h和复氧复糖3h的方法行OGD/R，采用缺氧10min复氧30 min且重复3次的方法行HPC。处理结束后采用MTT法测定细胞活力，测定细胞培养液LDH活性。结果与C组比较，OGD/R组、R＋OGD/R组、HPC＋OGD/R组、R＋HPC＋OGD/R组心肌细胞活力降低，细胞培养液LDH活性升高（ P＜0.05），R组上述指标差异无统计学意义（ P＞0.05）；与OGD/R组比较，HPC＋OGD/R组心肌细胞活力升高，细胞培养液LDH活性降低（ P＜0.05）；与HPC＋OGD/R组比较，R＋HPC＋OGD/R组心肌细胞活力降低，细胞培养液LDH活性升高（ P＜0.05）。结论 mTOR参与了HPC减轻乳鼠离体心肌细胞OGD/R损伤。
Objective To evaluate the role of mammalian target of rapamycin （mTOR ） in reduction of oxygen-glucose deprivation and restoration （OGD/R ）-induced injury to isolated cardiomyocytes by hypoxic preconditioning （HPC） in neonatal rats .Methods Primary cardiomyocytes were obtained from neonatal Sprague-Dawley rats （1-3 days after birth ） ,and seeded in 96-well plates at the density of 1 × 105 cells/ml （200μl/well ） . The cells were randomly divided into 6 groups （ n=6 each ） using a random number table：control group （group C） ,rapamycin group （group R） ,group OGD/R ,rapamycin＋OGD/R group （group R＋OGD/R）;group HPC＋OGD/R ,and rapamycin＋HPC＋OGD/R group （group R＋HPC＋OGD/R） .Rapamycin （final concentration 20 nmol/L） was added to the culture media and the cells were then incubated for 1 h in group R .In R＋OGD/R group ,rapamycin （final concentration 20 nmol/L ） was added to the culture media ,the cells were then incubated for 1 h ,and then OGD/R was performed .In HPC＋OGD/R group ,OGD/R was performed immediately after HPC was completed .In R＋HPC＋OGD/R group ,rapamycin （final concentration 20 nmol/L） was added to the culture media ,the cells were then incubated for 1 h , and then HPC and OGD/R were performed . All the cells were subjected to OGD for 2 h followed by restoration of O2-glucose supply for 3 h to induce OGD/R .HPC was induced by 3 cycles of 10 min hypoxia followed by 30 min reoxygenation .The cell viability （using MTT assay ） and lactic＆nbsp;dehydrogenase （LDH） activity in the culture medium were determined .Results Compared with group C ,the cell viability was significantly decreased , and LDH activities in the culture medium were increased in OGD/R , R＋OGD/R ,HPC＋OGD/R and R＋ HPC＋OGD/R groups （ P0.05） .The cell viability was significantly higher ,and LDH activity in the culture medium were lower in HPC＋OGD/R group than in OGD/R group ,and in R＋HPC＋OGD/R group than in HPC ＋ OGD/R group （ P〈 0.05 ） .Co
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2014年第3期363-365,共3页
Chinese Journal of Anesthesiology
基金
高等学校博士学科点专项科研基金联合资助课题（20126517110005）
新疆自治区重点学科项目[新教研（2010）7号]
