摘要
目的 探讨GR和HSP90在糖皮质激素敏感型(SS)、依赖型(SD)和抵抗型(SR)哮喘中的表达及其在SR哮喘发病中的作用。
方法 采用反转录-聚合酶链反应(RT-PCR)和蛋白印迹(Western blot)的方法,分别测定正常人(10例)、SS哮喘(10例)、SD哮喘(5例)和SR哮喘(10例)的外周血单个核细胞(PBMC)中,GR、HSP90mRNA表达和GR、HSP90蛋白水平,并在体外用IL-2、IL-4分别和联合刺激上述细胞,观察其受刺激后GR、HSP90mRNA表达和GR、HSP90蛋白水平的改变情况。
结果 SR哮喘的GR和HSP90mRNA表达水平最高(分别为0.730±0.171、1.122±0.165),SS哮喘次之(分别为0.359±0.350、0.885_+0.250),SD哮喘最低(分别为0.017_+0.008、0.078_+0.039)。正常人有一定表达(分别为0.052±0.013、0.362±0.101)。GR和HSP90mRNA的表达各组间相比P<0.05。正常人、SS、SD和SR哮喘HSP90/GR的比值分别为7.15±1.84、8.39±7.95、5.51±3.30、1.574±0.18,SR哮喘HSP90/GR比值明显低于前三组(P<0.05)。IL-2和IL-4单独刺激对SS、SD和SR哮喘的GR、HSP90mRNA表达无明显影响,二者联合刺激可使SS、SD和SR哮喘GRmRNA表达增强和SS、SD哮喘HSP90mRNA表达增强,却不能使SR哮喘HSP90mRNA表达增强。GR和HSP90的蛋白水平与其mRNA的表达相同。
结论 SR哮喘中HSP90表达相对不足造成HSP90/GR比值降低可能是SR哮喘形成的原因之一,IL-2+IL-4对GR和HSP90表达的不同调节作用可能是形成SR哮喘HSP90/GR比值降低的原因。
Objective To investigate the expression of GR,HSP90mRNA in peripheral blood mononuclear cells(PBMC) from steroid-sensitive asthma(SS),steroid-dependant asthma(SD) and steroid-resistant asthma(SR) and the role of GR and HSP90 on the pathogenesis of SR. Method With reverse transcription-polymerase chain reaction(RT-PCR) and Western blot ,the expression of GR and HSP90 were detected in PBMC from 10 normal volunteers, 10SS,5SD and 6SR as and as in vitro cultured in the absence and presence of IL-2 and IL-4. Results The expression of GR,HSP90mRNA in PBMC from SR patients(0.730± 0.171,1.122 ± 0.165 respectively) was the highest.The seconds from SS patients(0.359±0.350,0.885 ± 0.250 respectively).The lowest was from SD patients(0.017±0.008, 0.078±0.039 respectively).The expression of GR and HSP90mRNA in PBMC from normal volunteers was 0.052 ± 0.013 and 0.362±0.101 respectively.The difference of the expression of GR or HSP90 between each other of them was significant(P<0.05).The ratio of hsp90/GR in normal volunteers,SS,SD and SR patients was 7.15 ± 1.84,8.39 ± 7.95,5.51 ± 3.30 and 1.57 ± 0.18 respectively.HSP90/GR in SR was significant lower than that of the others and it suggested that the expression of HSP90mRNA was not sufficient in this patients.When PBMC from SS,SD and SR were incubated with IL-2 or IL-4 alone,no change of GR and HSP90 mRNA expression were observed. When incubated with combination of IL-2 and IL-4,a significant higher expression of GRmRNA expression was observed in all asthmatics and a significant higher expression of HSP90mRNA was observed only in SS and SD patients but not in SR patients.The level of proteins of GR and HSP90 was similar with that of their mRNA experssion.
Conclusion The low of the ratio of HSP90/GR may be one of the cause of SR.IL-2 and IL-4 may play a important role in the imbalance of HSP90/GR.
引文
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