CAJ | 학술논문

目的探讨NF-κB信号通路在异丙酚抑制脂多糖(LPS)诱导RAW264.7细胞诱导型一氧化氮合酶( iNOS)基因表达上调中的作用.方法体外培养RAW264.7细胞,以5×105/ml密度接种于6 cm培养皿(3 ml/皿)或6孔板(2 ml/孔),采用随机数字表法,将其随机分为3组(n=18):正常对照组(C组)、LPS组(L组)和LPS+异丙酚(LP组).C组不做任何处理；L组和LP组均加入1μg/mlLPS,LP组于加入LPS前2h加入50 μmol/L异丙酚.于LPS孵育30 min时,每组取6皿和6孔,收集细胞,分别采用免疫印迹法测定磷酸化IκB激酶(p-IKK)和NF-κB活性；于LPS孵育6h时,每组取6皿,收集细胞,测定iNOS mRNA表达.结果与C组比较,L组p-IKK和iNOS mRNA表达上调,NF-κB活性升高(P＜0.05)；与L组比较,LP组p-IKK和iNOS mRNA表达下调,NF-κB活性降低(P＜0.05).结论 NF-κB信号通路参与了异丙酚抑制脂多糖诱导的RAW264.7细胞iNOS基因表达上调.
목적 탐토NF-κB신호통로재이병분억제지다당(LPS)유도RAW264.7세포유도형일양화담합매( iNOS)기인표체상조중적작용.방법 체외배양RAW264.7세포,이5×105/ml밀도접충우6 cm배양명(3 ml/명)혹6공판(2 ml/공),채용수궤수자표법,장기수궤분위3조(n=18):정상대조조(C조)、LPS조(L조)화LPS+이병분(LP조).C조불주임하처리；L조화LP조균가입1μg/mlLPS,LP조우가입LPS전2h가입50 μmol/L이병분.우LPS부육30 min시,매조취6명화6공,수집세포,분별채용면역인적법측정린산화IκB격매(p-IKK)화NF-κB활성；우LPS부육6h시,매조취6명,수집세포,측정iNOS mRNA표체.결과 여C조비교,L조p-IKK화iNOS mRNA표체상조,NF-κB활성승고(P＜0.05)；여L조비교,LP조p-IKK화iNOS mRNA표체하조,NF-κB활성강저(P＜0.05).결론 NF-κB신호통로삼여료이병분억제지다당유도적RAW264.7세포iNOS기인표체상조.
Objective To investigate the role of nuclear factor-kappa B (NF-κB)signaling pathway in propofol-induced suppression of up-regulation of inducible nitric oxide synthase (iNOS) gene expression in LPSstimulated RAW264.7 cells.Methods RAW264.7 cells were purchased from cell bank of Chinese Academy of Sciences and cultured in DMEM culture medium containing 10％ fetal bovine serum.The cells were seeded in 6 cm diameter dishes (3 ml/dish) or in 6-well plates (2 ml/well) with a density of 5 × 105/ml and randomly divided into 3 groups ( n =18): normal control group (group C),group LPS (group L)and group LPS + propofol (group LP).The cells were incubated with LPS 1 μg/ml in groups L and LP.Propofol 50μmol/L was added to the culture medium at 2 h before LPS in group LP.Cells were harvested at 30 min after being stimulated with LPS.Phosphorylation of IκB kinase(p-IKK) and NF-κB activity were detected by Western blot.The expression of iNOS mRNA was determined after 6 h exposure of the cells to LPS.Results LPS significantly up-regulated the expression of p-IKK and iNOS mRNA and increased NF-κB activity in group L as compared with group C.Propofol pretreatment significantly attenuated the effects of LPS on p-IKK,iNOS mRNA expression and NF-κB activity.Conclusion NF-κB signaling pathway is involved in the propofol-induced suppression of up-regulation of iNOS mRNA expression in LPS-stimulated RAW264.7 cells.