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目的:探讨磷酸二酯酶(PDE)抑制剂对慢性阻塞性肺疾病(COPD)患者外周血单个核细胞(PBMC)IL-8mRNA表达的影响及机制。方法:采集20例急性发作期COPD患者(A组)和15例正常人(B组)的PBMC。A组的每份PBMC分4组培养:A1组为空白对照,A2组和A3组分别加入100μmol/L及1mmol/L的非选择性PDE抑制剂茶碱,A4组加入选择性PDEⅣ抑制剂Rolipram。应用RT-PCR检测IL-8mRNA的表达,以免疫组化法检测NF-κB阳性细胞的百分率,用Westernblot检测I-κB蛋白的含量。结果:(1)A1组PB-MC中IL-8mRNA的表达及NF-κB阳性细胞的百分率,均显著高于B组;IκB的水平显著低于B组(P<0.01)。(2)与A1组比较,A2组和A3组的PB-MC中IL-8mRNA的表达无显著差异(P>0.05);A4组IL-8mRNA的表达明显下降(P<0.01)。(3)与A1组比较,A2组NFκB阳性细胞的百分率及IκB的水平无显著差异(P>0.05);而A3组的PBMC中NFκB阳性细胞的百分率下降(P<0.05),IκB蛋白的水平无显著变化(P>0.05)。A4组PB-MC中,NF-κB阳性细胞的百分率较A1组明显下降,I-κB蛋白的水平明显增加(P<0.01)。结论:COPD患者PBMC中IL-8mRNA的表达较正常增高,提示IL-8参与了COPD的发病过程;选择性PDEⅣ抑制剂Roli-pram可抑制IL-8基因的表达,其作用机制可能与NF-κB阳性细胞百分率的下降有关。非选择性PDE抑
Objective: To investigate the effect and mechanism of phosphodiesterase (PDE) inhibitor on the expression of IL-8mRNA in peripheral blood mononuclear cells (PBMCs) in patients with chronic obstructive pulmonary disease (COPD). Methods: PBMC from 20 patients with acute exacerbation of COPD (group A) and 15 normal subjects (group B) were collected. Each PBMC in group A was cultured in 4 groups: group A1 was blank control, group A2 and group A3 were added 100μmol / L and 1mmol / L non-selective PDE inhibitor theophylline, A4 group added selective PDE Ⅳ inhibitor Rolipram . The expression of IL-8 mRNA was detected by RT-PCR. The percentage of NF-κB positive cells was detected by immunohistochemistry. The content of I-κB protein was detected by Western blot. Results: (1) The expression of IL-8 mRNA and the percentage of NF-κB positive cells in PB-MC in A1 group were significantly higher than those in B group; the level of IκB was significantly lower than that in B group (P <0.01). (2) Compared with group A1, the expression of IL-8mRNA in PB-MC in A2 group and A3 group had no significant difference (P> 0.05). The expression of IL-8mRNA in A4 group was significantly decreased (P <0.01). (3) Compared with A1 group, the percentage of NFκB positive cells in A2 group and the level of IκB had no significant difference (P> 0.05); while the percentage of NFκB positive cells in PBMC of A3 group decreased (P <0.05) No significant change (P> 0.05). The percentage of NF-κB positive cells in A4-PBMCs was significantly lower than that in A1 and the level of I-κB protein was significantly increased (P <0.01). Conclusion: The expression of IL-8mRNA in PBMC of COPD patients is higher than normal, suggesting that IL-8 is involved in the pathogenesis of COPD. Roli-pram, a selective PDEⅣinhibitor, can inhibit the expression of IL-8, κB-positive cells decreased. Non-selective PDE inhibition