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.