CAJ | 학술논문

目的探讨微小RNA-141(miR-141)对人单核细胞株THP-1高迁移率族蛋白B1(HMGB1)表达的调控作用.方法体外培养THP-1细胞,化学合成人miR-141的拟似物和抑制剂.用脂质体Lipofectamine RNAi MAX转染miR-141的拟似物或抑制剂进入THP-1细胞,转染48 h后分别用实时荧光定量逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹试验(Western blotting)检测HMGB1的mRNA和蛋白表达.结果转染100 nmol/L miR-141拟似物后,可提高THP-1细胞miR-141的表达水平(35.33±7.24比1.21±0.20,t=-8.408,P=0.010);转染100 nmol/LmiR-141抑制剂后,可抑制THP-1细胞miR-141的表达水平(0.55±0.12比1.09±0.05,t=7.473,P=0.002).与相应对照组比较,THP-1细胞在过表达miR-141后,HMGB1的mRNA和蛋白表达明显下调(mRNA:0.43±0.06比0.97±0.08,t=9.760,P=0.001;蛋白:0.63±0.12比1.00±0.11,t=2.991,P=0.040);而抑制THP-1细胞中miR-141后,HMGB1的mRNA和蛋白表达均明显上调(mRNA:2.13±0.11比1.16±0.13,t=-9.977,P=0.001;蛋白:1.78±0.04比0.96±0.09,t=-13.778,P=0.000).结论 miR-141能调控人单核细胞HMGB1的表达水平,提示miR-141在调控免疫细胞的炎症反应过程中具有重要的作用.
목적 탐토미소RNA-141(miR-141)대인단핵세포주THP-1고천이솔족단백B1(HMGB1)표체적조공작용.방법 체외배양THP-1세포,화학합성인miR-141적의사물화억제제.용지질체Lipofectamine RNAi MAX전염miR-141적의사물혹억제제진입THP-1세포,전염48 h후분별용실시형광정량역전록-취합매련반응(RT-PCR)화단백질면역인적시험(Western blotting)검측HMGB1적mRNA화단백표체.결과 전염100 nmol/L miR-141의사물후,가제고THP-1세포miR-141적표체수평(35.33±7.24비1.21±0.20,t=-8.408,P=0.010);전염100 nmol/LmiR-141억제제후,가억제THP-1세포miR-141적표체수평(0.55±0.12비1.09±0.05,t=7.473,P=0.002).여상응대조조비교,THP-1세포재과표체miR-141후,HMGB1적mRNA화단백표체명현하조(mRNA:0.43±0.06비0.97±0.08,t=9.760,P=0.001;단백:0.63±0.12비1.00±0.11,t=2.991,P=0.040);이억제THP-1세포중miR-141후,HMGB1적mRNA화단백표체균명현상조(mRNA:2.13±0.11비1.16±0.13,t=-9.977,P=0.001;단백:1.78±0.04비0.96±0.09,t=-13.778,P=0.000).결론 miR-141능조공인단핵세포HMGB1적표체수평,제시miR-141재조공면역세포적염증반응과정중구유중요적작용.
Objective To investigate the regulatory effect of microRNA-141(miR-141)on expression of high mobility group protein B1(HMGB1)in human monocytes THP-1 cell line.Methods THP-1 cells were transfected with miR-141 mimic or inhibitor(100 nmol/L)for 48 hours with lipofectamine RNAi MAX.The levels of miR-141 and HMGB1 mRNA in the THP-1 cells were detected by real-time fluorescence quantitation reverse transcription-polymerase chain reaction(RT-PCR),and HMGB1 protein was determined with Western blotting.Results The levels ofmiR-141 could be up regulated(35.33±7.24 vs.1.21±0.20,t=-8.408,P=0.010)or down regulated(0.55±0.12 vs 1.09±0.05,t=7.473,P=0.002)after being transfected with 100 nmol/L miR-141 mimic or inhibitor for 48 hours by lipofectamine RNAi MAX in THP-1,and the level of HMGB1 mRNA and protein decreased (mRNA:0.43±0.06 vs.0.97±0.08,t=9.760,P=0.001;protein:0.63±0.12 vs.1.00±0.11,t=2.991,P=0.040)or increased(mRNA:2.13±0.11 vs.1.16±0.13,t=-9.977,P=0.001;protein:1.78±0.04 vs.0.96±0.09,t=-13.778,P=0.000)simultaneously compared with the control group.Conclusion miR-141 is involved in regulation of inflammation through HMGB1 gene and protein pathway,suggesting that miR-141 plays an important role in regulating immune cells during the inflammatory response.