CAJ | 학술논문

目的探讨p38丝裂原活化蛋白激酶(p38 MAPK)抑制剂SB203580对糖皮质激素敏感性的影响机制.方法在体内实验中,Wistar大鼠60只随机分为正常对照组、哮喘组、烟雾暴露哮喘组及SB203580干预组各15只.经荧光定量PCR检测大鼠肺组织糖皮质激素受体(GR)、热休克蛋白90( HSP90)和p38 MAPK mRNA的表达,经Western印迹检测大鼠肺组织GR、HSP90和p38MAPK蛋白的表达.在体外实验中,培养人肺腺癌A549细胞,将其平均分为4组并进行如下处理:A组:空白对照组；B组:加用地塞米松(1μmol/L)；C组:加用地塞米松+尼古丁(均为1μmol/L)；D组:加用地塞米松+尼古丁+ SB203580(均为1μmol/L).采用免疫荧光染色技术分析GR亚细胞定位.结果烟雾暴露哮喘组和SB203580干预组大鼠肺组织GR mRNA表达分别为0.671 ±0.002和0.595 ±0.061 (P =0.065),GR蛋白表达分别为0.700±0.033和0.628 ±0.091(P =0.148).两组大鼠肺组织HSP90 mRNA表达分别为0.558±0.009和0.377 ±0.046(P =0.000),HSP90蛋白表达分别为0.507±0.030和0.402±0.050(P=0.005)；p38 MAPK mRNA表达分别为0.971±0.012和0.278±0.049(P =0.000),p38 MAPK蛋白表达分别为0.982±0.038和0.338±0.042(P =0.000).C组A549细胞GR的核质比(0.077±0.047)明显低于D组(0.592±0.249)(P=0.000).结论 p38 MAPK抑制剂SB203580通过促使GR核转位提高糖皮质激素敏感性.
목적 탐토p38사렬원활화단백격매(p38 MAPK)억제제SB203580대당피질격소민감성적영향궤제.방법 재체내실험중,Wistar대서60지수궤분위정상대조조、효천조、연무폭로효천조급SB203580간예조각15지.경형광정량PCR검측대서폐조직당피질격소수체(GR)、열휴극단백90( HSP90)화p38 MAPK mRNA적표체,경Western인적검측대서폐조직GR、HSP90화p38MAPK단백적표체.재체외실험중,배양인폐선암A549세포,장기평균분위4조병진행여하처리:A조:공백대조조；B조:가용지새미송(1μmol/L)；C조:가용지새미송+니고정(균위1μmol/L)；D조:가용지새미송+니고정+ SB203580(균위1μmol/L).채용면역형광염색기술분석GR아세포정위.결과 연무폭로효천조화SB203580간예조대서폐조직GR mRNA표체분별위0.671 ±0.002화0.595 ±0.061 (P =0.065),GR단백표체분별위0.700±0.033화0.628 ±0.091(P =0.148).량조대서폐조직HSP90 mRNA표체분별위0.558±0.009화0.377 ±0.046(P =0.000),HSP90단백표체분별위0.507±0.030화0.402±0.050(P=0.005)；p38 MAPK mRNA표체분별위0.971±0.012화0.278±0.049(P =0.000),p38 MAPK단백표체분별위0.982±0.038화0.338±0.042(P =0.000).C조A549세포GR적핵질비(0.077±0.047)명현저우D조(0.592±0.249)(P=0.000).결론 p38 MAPK억제제SB203580통과촉사GR핵전위제고당피질격소민감성.
Objective To establish a model of cigarette smoke exposure to asthmatic rats and glucocorticoid resistance induced by nicotine in alveolar epithelioid cells A549 and study the mechanism for the change of glucocorticoid sensitivity induced by p38 mitogen-activated protein kinase (p38 MAPK )inhibitor SB203580. Methods Sixty Wistar rats were randomly divided into 4 groups:normal group,asthmatic group,cigarette smoke exposure to asthmatic group and SB203580 group.The mRNA expressions of glucocorticoid receptor (GR),heat shock protein 90 (HSP90) and p38 MAPK were detected by real time-polymerase chain reaction (RT-PCR) while their protein expressions detected by Western blot in vivo.A549 cells were divided averagely into 4 groups:group A:normal; group B:1 μmol/L dexamethasone (DEX) ; group C:1 μmol/L DEX + 1 μ mol/L nicotine; group D:1 μmol/L DEX + 1 μmol/L nicotine +1 μ mol/L SB203580. Immunofluorescence staining was used to study the in vitro colocalization ofglucocorticoid receptor (GR) in A549 cells.Results The mRNA expression of GR was 0.671 ± 0.002 in cigarette smoke exposure to asthmatic group and 0.595 ± 0.061 in SB203580 group ( P =0.065 ). The protein expression of GR was 0.700 ± 0.033 in cigarette smoke exposure to asthmatic group and 0.628 ±0.091 in SB203580 group (P =0.148).The mRNA expression of HSP90 was 0.558 ± 0.009 in cigarette smoke exposure to asthmatic group and 0.377 ± 0.046 in SB203580 group (P =0.000 ). The protein expression of HSP90 was 0.507 ± 0.030 in cigarette smoke exposure to asthmatic group and 0.402 ± 0.050in SB203580 group ( P =0.005 ). The mRNA expression of p38 MAPK was 0.971 ± 0.012 in cigarette smoke exposure to asthmatic group and 0.278 ± 0.049 in SB203580 group (P =0.000). The proteinexpression of p38 MAPK was 0.982 ± 0.038 in cigarette smoke exposure to asthmatic group and 0.338 ±0.042 in SB203580 group ( P =0.000 ).The ratio of GR amount within A549 nucleus versus that in cytoplasm was 0.077 ± 0.047 in group C and 0.592 ± 0.249 in group D ( P =0.000 ).Conclusion The mechanism of SB203580 enhancing the corticosteroid sensitivity may be improving nuclear translocation of GR to elevate corticosteroid sensitivity.