PDF(Pubmed)
Abstract：
OBJECTIVE: The response to glucocorticoids is hampered in many COPD patients by a yet unknown mechanism. Earlier we reported that short-term heat exposure of primary human bronchial epithelial cells (BEC) and airway smooth muscle cells (ASMC) of asthma patients increased the expression and secretion of extracellular heat shock proteins (eHSPs) resulting in increased expression of glucocorticoid receptor (GR) in BEC and inhibition of ASMC remodeling. The aim of the present study was to assess if the same mechanism is also present in primary airway wall cells of COPD patients.
METHODS: Primary BEC and ASMC were established from endobronchial biopsies obtained from COPD patients (n = 73), who participated in the HISTORIC study, an investigator-initiated and driven clinical trial. Secretion and protein expression of HSPs was assessed by ELISA and Western blotting. Expression of total GR, its isoforms GRα and GRβ and toll-like receptor 4 (TLR4) was determined by Western-blotting.
RESULTS: Short heat exposure (65 °C, 10 s) of BEC resulted in a significant increase of the secretion of eHSP70 and eHSP90, while the intracellular protein was not altered. Heat treatment or exposure to eHSP70 or eHSP90 had no effect on the expression of GR and GR-isoforms. However, eHSP70 and eHSP90 significantly reduced the expression of TLR4.
CONCLUSIONS: The results of this study indicate that primary airway cells from COPD patients respond differently to heat exposure and extracellular HSP70 or HSP90 than cells from asthma patients regarding the expression of GR and this may explain the reduced response to glucocorticoids in patients with COPD.
BACKGROUND: ISRCTN11017699.
METHODS: Primary BEC and ASMC were established from endobronchial biopsies obtained from COPD patients (n = 73), who participated in the HISTORIC study, an investigator-initiated and driven clinical trial. Secretion and protein expression of HSPs was assessed by ELISA and Western blotting. Expression of total GR, its isoforms GRα and GRβ and toll-like receptor 4 (TLR4) was determined by Western-blotting.
RESULTS: Short heat exposure (65 °C, 10 s) of BEC resulted in a significant increase of the secretion of eHSP70 and eHSP90, while the intracellular protein was not altered. Heat treatment or exposure to eHSP70 or eHSP90 had no effect on the expression of GR and GR-isoforms. However, eHSP70 and eHSP90 significantly reduced the expression of TLR4.
CONCLUSIONS: The results of this study indicate that primary airway cells from COPD patients respond differently to heat exposure and extracellular HSP70 or HSP90 than cells from asthma patients regarding the expression of GR and this may explain the reduced response to glucocorticoids in patients with COPD.
BACKGROUND: ISRCTN11017699.
摘要：
目的：许多COPD患者对糖皮质激素的反应受到一种未知机制的阻碍。早期我们报道了哮喘患者原代人支气管上皮细胞（BEC）和气道平滑肌细胞（ASMC）的短期热暴露会增加细胞外热休克蛋白（eHSPs）的表达和分泌，从而导致糖皮质激素受体（GR）在BEC中的表达增加并抑制ASMC重塑。本研究的目的是评估COPD患者的原发性气道壁细胞中是否也存在相同的机制。
方法：从COPD患者（n=73）的支气管活检中建立原发性BEC和ASMC，参加历史研究的人，研究者发起和驱动的临床试验。通过ELISA和Western印迹评估HSPs的分泌和蛋白表达。总GR的表达式,通过Western印迹测定其亚型GRα和GRβ以及toll样受体4（TLR4）。
结果：短暂的热暴露（65°C，10s）的BEC导致eHSP70和eHSP90的分泌显着增加，而细胞内蛋白没有改变。热处理或暴露于eHSP70或eHSP90对GR和GR亚型的表达没有影响。然而,eHSP70和eHSP90显著降低TLR4的表达。
结论：本研究结果表明，COPD患者的原发性气道细胞对热暴露和胞外HSP70或HSP90的反应与哮喘患者的细胞对GR表达的反应不同，这可能解释了COPD患者对糖皮质激素反应降低的原因。
背景：ISRCTN11017699。
方法：从COPD患者（n=73）的支气管活检中建立原发性BEC和ASMC，参加历史研究的人，研究者发起和驱动的临床试验。通过ELISA和Western印迹评估HSPs的分泌和蛋白表达。总GR的表达式,通过Western印迹测定其亚型GRα和GRβ以及toll样受体4（TLR4）。
结果：短暂的热暴露（65°C，10s）的BEC导致eHSP70和eHSP90的分泌显着增加，而细胞内蛋白没有改变。热处理或暴露于eHSP70或eHSP90对GR和GR亚型的表达没有影响。然而,eHSP70和eHSP90显著降低TLR4的表达。
结论：本研究结果表明，COPD患者的原发性气道细胞对热暴露和胞外HSP70或HSP90的反应与哮喘患者的细胞对GR表达的反应不同，这可能解释了COPD患者对糖皮质激素反应降低的原因。
背景：ISRCTN11017699。
