CAJ | 학술논문

目的评价异丙酚预处理对缺氧复氧诱发HEPG2细胞内质网应激的影响.方法 HEPG2细胞采用随机数字表法分为4组(n=6):对照组(C组)常规培养42 h;异丙酚组(P组)用含异丙酚(终浓度10 μmol/L)培养基孵育6h,更换正常培养基培养36 h;缺氧复氧组(H/R组):常规培养6h,缺氧24h复氧12 h;异丙酚预处理组(PP组)用含异丙酚(终浓度10 μmol/L)培养基孵育6h,更换正常培养基置入三气培养箱(1％O2、5％ CO2和94％N2)缺氧24h,随后放入正常培养箱培养12 h.采用MTT比色法检测细胞活力,采用Western blot法检测免疫球蛋白重链结合蛋白(BIP)、C/EBP同源蛋白(CHOP)和活化型caspase-3的蛋白表达,采用RT-PCR法检测BIP、CHOP和caspase-3的mRNA表达.结果与C组比较,H/R组和PP组细胞活力降低,BIP、CHOP、活化型caspase-3的蛋白及其mRNA表达上调(P＜0.05),P组上述指标差异无统计学意义(P＞0.05);与H/R组比较,PP组细胞活力升高,BIP、CHOP、活化型caspase-3的蛋白及其mRNA表达下调(P＜0.05).结论异丙酚预处理可通过抑制内质网应激,促进细胞增殖,减轻HEPG2细胞缺氧复氧损伤.
목적 평개이병분예처리대결양복양유발HEPG2세포내질망응격적영향.방법 HEPG2세포채용수궤수자표법분위4조(n=6):대조조(C조)상규배양42 h;이병분조(P조)용함이병분(종농도10 μmol/L)배양기부육6h,경환정상배양기배양36 h;결양복양조(H/R조):상규배양6h,결양24h복양12 h;이병분예처리조(PP조)용함이병분(종농도10 μmol/L)배양기부육6h,경환정상배양기치입삼기배양상(1％O2、5％ CO2화94％N2)결양24h,수후방입정상배양상배양12 h.채용MTT비색법검측세포활력,채용Western blot법검측면역구단백중련결합단백(BIP)、C/EBP동원단백(CHOP)화활화형caspase-3적단백표체,채용RT-PCR법검측BIP、CHOP화caspase-3적mRNA표체.결과 여C조비교,H/R조화PP조세포활력강저,BIP、CHOP、활화형caspase-3적단백급기mRNA표체상조(P＜0.05),P조상술지표차이무통계학의의(P＞0.05);여H/R조비교,PP조세포활력승고,BIP、CHOP、활화형caspase-3적단백급기mRNA표체하조(P＜0.05).결론 이병분예처리가통과억제내질망응격,촉진세포증식,감경HEPG2세포결양복양손상.
Objective To evaluate the effect of propofol preconditioning on endoplasmic reticulum stress induced by hypoxia-reoxygenation (H/R) in HEPG2 cells.Methods HEPG2 cells were randomly divided into 4 groups using a random number table:control group (group C),propofol group (group P),H/R group and H/R + propofol preconditioning group (group PP).In group C,the cells were cultured routinely for 42 h.In group H/R,after being cultured routinely for 6 h,the cells were exposed to 1％ O2 + 5％ CO2 + 94％ N2 for 12 h followed by 12 h reoxygenation.In group PP,the cells were cultured for 6 h in the culture medium containing propofol 10 μmol/L (final concentration),and then H/R was induced.The cell viability was detected by MTT assay.The expression of immunoglobulin heavy chain-binding protein (BIP),C/EBP homologous protein (CHOP) and activated caspase-3 was determined by Western blot.The expression of BIP,CHOP and caspase-3 mRNA was determined by RT-PCR.Results Compared with group C,the cell viability was significantly decreased,and the expression of BIP,CHOP and activated caspase-3 protein and mRNA was up-regnlated in H/R and PP groups,and no significant changes were found in the parameters mentioned above in group P.Compared with group H/R,the cell viability was significantly increased,and the expression of BIP,CHOP and activated caspase-3 protein and mRNA was down-regulated in PP group.Conclusion Propofol preconditioning can promote the cell proliferation and attenuate H/R injury to HEPG2 cells through inhibiting endoplasmic reticulum stress.