摘要
目的评价氢气对脂多糖（LPS）诱导人脐静脉内皮细胞凋亡的影响。方法离体培养人脐静脉内皮细胞株HUVEC-12，以1×104／ml接种于96孔培养板（每孑L200m）或以1×106／ml接种于6孔培养板（每孔2ml）。采用随机数字表法，将细胞随机分为4组（n=30）;正常对照组（C组）、氢气组（H2组）、LPS组和LPS＋H2组。C组和LPS组用正常培养基培养；H2组和LPS＋巩组用含饱和氢气的培养基培养，同时LPS组和LPS＋H2组加入LPS1μg/ml，而C组和地组加入等容量的生理盐水。孵育24h后用MrrT法检测细胞活力，流式细胞术测定细胞凋亡情况；采用ELISA法测定细胞上清液中HMGBl浓度。结果与C组和H2组比较，LPS组和LPS＋H2组细胞活力降低，细胞凋亡率和细胞上清液中HMGBl浓度升高（P〈O．05）；与LPS组比较，LPS＋H2组细胞活力升高，细胞凋亡率和细胞上清液中HMGBl浓度降低（P〈0．05）。结论氢气可有效减少LPS诱导人脐静脉内皮细胞凋亡，其机制与抑制HMGBl释放有关。
Objective To investigate the effect of hydrogen gas on lipopolysaccharide （LPS）-indnced apoptosis in human umbilical vein endothelial cells （HUVECs） in vitro. Methods HUVEC-12 cells were seeded in 96-well plates with a density of 1 × 104/ml （200 /A/hole） or in 6-well plates （2 ml/hole） with a density of 1×106/ml and randomly divided into 4 groups （.n = 30 each） ： control group （group C）, hydrogen gas （H2） group, LPS group and LPS ＋ H2 group. The cells were cultured in the plain culture medium in groups C and LPS or in hydrogen-saturated c61ture medium in groups H2 and LPS ＋ H2 . In addition, LPS 1 μg/ml was added simulta- neously in groups LPS and LPS ＋ H2 and the equal volume of normal saline was added instead in groups C and H2. The cell viability and apoptosis were measured by MTI＂ assay and flow cytometry respectively after 24 h incubation. The concentration of high-mobility group box 1 （ HMGB1 ） in the supernatant was determined by ELISA. Results Compared with groups C and H2, the cell viability was significantly decreased, and the apoptotic rate and concen- tration of HMGB1 in the supematant were significantly increased in groups LPS and LPS ＋ H2 （P 〈 0.05）. Com- pared with group LPS, the cell viability was significantly increased, and the apoptotic rate and concentration of HMGB1 in the supernatant were significantly decreased in group LPS ＋ H2 （ P 〈 0.05 ）. Conclusion Hhydrogen gas can effectively reduce LPS-induced apoptosis in HUVECs through inhibiting the release of HMGB1.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2012年第8期973-975,共3页
Chinese Journal of Anesthesiology
基金
国家自然科学基金（81071533,81101409）
天津市应用基础及前沿技术研究计划（11JCYBJC12900）
天津市卫生局资助项目（2011KZ108）
关键词
氢
脂多糖类
脐静脉
内皮细胞
细胞凋亡
Hydrogen
Lipopolysaccharides
Umbilical veins
Endothelial cells
Apoptosis
