摘要
目的评价异丙酚对人肝细胞氧糖剥夺-复氧复糖时自噬的影响。方法HL-7702细胞培养至对数生长期后，接种于培养板中，采用随机数字表法分为3组（n=12）：对照组（C组）、氧糖剥夺一复氧复糖组（OGD／R组）和异丙酚＋氧糖剥夺-复氧复糖组（P＋OGD／R组）。C组正常培养；OGD／R组和P＋OGD／R组氧糖剥夺6h，复氧复糖12h制备氧糖剥夺-复氧复糖损伤模型；P＋OGD／R组于缺氧缺糖前10min加入终浓度50μmol／L的异丙酚。采用MTT法检测细胞活力；采用Western blot法测定自噬蛋白微管相关蛋白1轻链3（LC3）和Beclin-1的表达水平；采用免疫荧光法观察自噬小体的数量及分布情况。结果与C组比较，OGD／R组和P＋OGD／R组肝细胞活力降低，LC3和Beclin-1的表达上调（P〈0．05），自噬小体数量增加；与OGD／R组比较，P＋OGD／R组肝细胞活力升高，LC3和Beclin—1的表达下调（P〈0．05），自噬小体数量减少。结论异丙酚减轻人肝细胞氧糖剥夺-复氧复糖损伤的机制可能与抑制自噬有关。
Objective To evaluate the effect of propofol on autophagy during oxygen-glucose deprivation and restoration （OGD/R） in human liver cells. Methods Human hepatic HL-7702 cells at the logarithmic growth phase were seeded into culture plates and randomly divided into 3 groups （n = 12 each） using a random number table： control group （ group C） , OGD/R group, and propofol ＋ OGD/R group （ group P＋OGD/R）. The cells were cultured in normal culture medium in group C. In OGD/R and P＋OGD/R groups, the cells were subjected to O2-glucose deprivation for 6 h followed by restoration of O2-glucose supply for 12 h. Propofol with a final concentration of 50 mmol/L was added at 10 rain before oxygen-glucose deprivation. The cell viability was detected by MTT assay. The expression of autophagy-related proteins such as microtubule-associated protein light chain 3 （LC3） and Beelin-1 was evaluated by Western blot. Immunofluorescence was used to determine the number and distribution of autophagosomes. Results Compared with group C, the cell viability was significantly decreased, the expression of LC3 and Beelin-1 was significantly up-regulated （P〈0.05）, and the number of autophagosomes was significantly increased in OGD/R and P＋OGD/R groups. Compared with group OGD/R, the cell viability was significantly increased, the expression of LC3 and Beclin-I was significantly down-regulated （P〈0.05） , and the number of autoph- agosomes was significantly decreased in group P＋OGD/R. Conclusion The mechanism by which propofol reduces OGD/R injury is probably related to inhibition of autophagy in human liver ceils.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2016年第6期725-727,共3页
Chinese Journal of Anesthesiology
基金
高等学校博士学科点专项科研基金（20133420110009）
关键词
异丙酚
再灌注损伤
肝
自噬
Propofol
Reperfusion injury
Liver
Autophagy
