CAJ | 학술논문

目的观察热休克蛋白70抑制剂PFTμ对脂多糖(LPS)诱导的RAW 264.7细胞一氧化氮(NO)的生成及诱生型一氧化氮合酶(iNOS)表达的作用以及与缺血再灌注损伤小鼠炎症反应的关系,并探讨其作用机制.方法采用LPS诱导的RAW 264.7细胞株建立细胞炎症反应模型,分为PFTμ组(PFTμ 20 μmol/L预处理15 min后加入LPS 2 g/L)和对照组(等量DMSO预处理15 min后加入等量LPS).Griess试剂法测定 NO释放量.Western blot法测定蛋白的表达.逆转录聚合酶链反应分析iNOS mRNA表达改变.建立小鼠心脏缺血再灌注模型,分为PFTμ组(40 mg/kg,溶于DMSO腹腔注射)和对照组(等量DMSO腹腔注射),测定小鼠心肌梗死面积的变化.结果 PFTμ可抑制LPS诱导的RAW264.7细胞NO的释放(PFTμ组NO的释放显著低于对照组,P<0.05).PFTμ可下调LPS诱导的RAW264.7细胞iNOS mRNA和蛋白表达(PFTμ组iNOS mRNA和蛋白表达均显著低于对照组,P均<0.05).PFTμ可减少缺血再灌注小鼠心脏梗死面积(PFTμ组小鼠心肌梗死面积显著低于对照组,P<0.05).结论 PFTμ有抑制炎症反应的作用,该作用机制可能与抑制NO的生成有关.
목적 관찰열휴극단백70억제제PFTμ대지다당(LPS)유도적RAW 264.7세포일양화담(NO)적생성급유생형일양화담합매(iNOS)표체적작용이급여결혈재관주손상소서염증반응적관계,병탐토기작용궤제.방법 채용LPS유도적RAW 264.7세포주건립세포염증반응모형,분위PFTμ조(PFTμ 20 μmol/L예처리15 min후가입LPS 2 g/L)화대조조(등량DMSO예처리15 min후가입등량LPS).Griess시제법측정 NO석방량.Western blot법측정단백적표체.역전록취합매련반응분석iNOS mRNA표체개변.건립소서심장결혈재관주모형,분위PFTμ조(40 mg/kg,용우DMSO복강주사)화대조조(등량DMSO복강주사),측정소서심기경사면적적변화.결과 PFTμ가억제LPS유도적RAW264.7세포NO적석방(PFTμ조NO적석방현저저우대조조,P<0.05).PFTμ가하조LPS유도적RAW264.7세포iNOS mRNA화단백표체(PFTμ조iNOS mRNA화단백표체균현저저우대조조,P균<0.05).PFTμ가감소결혈재관주소서심장경사면적(PFTμ조소서심기경사면적현저저우대조조,P<0.05).결론 PFTμ유억제염증반응적작용,해작용궤제가능여억제NO적생성유관.
Objective To observe the effects of hot shock protein 70 (HSP70) inhibitor (PFTμ) on inflammation response in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and mice underwent myocardial ischemia-reperfusion (I/R) injury.Methods RAW264.7 macrophage line of mice was stimulated by LPS as an inflammatory model. These were divided into control (15 min DMSO pretreatment and LPS 2 g/L)and PFTμ treated groups(15 min PFTμ 20 μmol/L pretreatment and LPS 2 g/L). NO concentration was measured by Griess Kit. The expression of iNOS protein and mRNA were detected by Western blot and RT-PCR.Infarct size was determined on mice underwent myocardial ischemia-reperfusion (I/R) injury in the absence or presence (PFTμ 40 mg/kg,intraperitoneal injection).Results PFTμ significantly blocked the production of NO and protein and mRNA expression of iNOS (P<0.05 vs. control). PFTμ also significantly reduced the infarct size on mice underwent I/R injury (P<0.05 vs. control).Conclusion These results suggest that PFTμ could be a potential therapeutic agent for the treatment of inflammatory diseases through inhibiting the production of NO and reducing informatory responses.