CAJ | 학술논문

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(每孔200μl)或以1 × 106/ml接种于6孔培养板(每孔2 ml).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+ H2组用含饱和氢气的培养基培养,同时LPS组和LPS+H2组加入LPS 1μg/ml,而C组和H2组加入等容量的生理盐水.孵育24 h后用MTT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HMGB1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HMGB1浓度升高(P＜0.05)；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HMGB1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HMGB1释放有关.
목적 평개경기대지다당(LPS)유도인제정맥내피세포조망적영향.방법 리체배양인제정맥내피세포주HUVEC-12,이1×104/ml접충우96공배양판(매공200μl)혹이1 × 106/ml접충우6공배양판(매공2 ml).채용수궤수자표법,장세포수궤분위4조(n=30):정상대조조(C조)、경기조(H2조)、LPS조화LPS+H2조.C조화LPS조용정상배양기배양；H2조화LPS+ H2조용함포화경기적배양기배양,동시LPS조화LPS+H2조가입LPS 1μg/ml,이C조화H2조가입등용량적생리염수.부육24 h후용MTT법검측세포활력,류식세포술측정세포조망정황；채용ELISA법측정세포상청액중HMGB1농도.결과 여C조화H2조비교,LPS조화LPS+ H2조세포활력강저,세포조망솔화세포상청액중HMGB1농도승고(P＜0.05)；여LPS조비교,LPS+H2조세포활력승고,세포조망솔화세포상청액중HMGB1농도강저(P＜0.05).결론 경기가유효감소LPS유도인제정맥내피세포조망,기궤제여억제HMGB1석방유관.
Objective To investigate the effect of hydrogen gas on lipopolysaccharide (LPS)-induced 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 μl/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 culture medium in groups H2 and LPS + H2.In addition,LPS 1 μg/ml was added simultaneously 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 MTT 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 concentration of HMGB1 in the supernatant were significantly increased in groups LPS and LPS + H2 (P ＜ 0.05).Compared 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.