成纤维细胞对纤维胶原蛋白组装的缺陷促进哮喘气道重塑
2019/04/19
摘要
基本原理：组织学染色已被用作可视化与哮喘气道重塑相关的细胞外基质（ECM）变化的金标准，但它们没有提供关于ECM的生化和结构特征的信息，而这对于理解组织功能的改变是至关重要的。
目的：展示使用非线性光学显微镜（NLOM）和纹理分析算法来成像原纤维胶原（二次谐波产生）和弹性蛋白（双光子激发自发荧光），以获得关于哮喘重塑ECM环境的生化和结构信息。
方法：来自哮喘（n = 13）和对照（n = 12）供体的非移植供体肺，通过NLOM评估气道胶原和弹性蛋白纤维，以及体外实验的肺成纤维细胞的提取。
检测和主要结果：与对照组相比，在哮喘组大、小气道内，使用NLOM成像的纤维状胶原蛋白不仅增加，而且高度混乱、碎裂。此外，这种结构改变存在于儿科和成人哮喘供体中，与致命疾病无关。体外研究表明，哮喘气道成纤维细胞对纤维胶原蛋白-I的组装有缺陷，对胶原纤维组装很重要的核心蛋白聚糖表达较少。使用透射电子显微镜观察发现，与对照相比，在哮喘气道中胶原原纤维的组装更加混乱。
结论：NLOM成像使得ECM的结构评估成为可能，并且数据表明哮喘中的气道重塑包括气道成纤维细胞引起的紊乱纤维胶原的不断积聚。该研究强调了NLOM用来评估体内气道重塑的潜在临床应用价值。
Defective Fibrillar Collagen Organization by Fibroblasts Contributes to Airway Remodeling in Asthma.
Mostaço-Guidolin LB, Osei ET, Ullah J, Hajimohammadi S, Fouadi M, Li X, Li V, Shaheen F, Yang CX, Chu F, Cole DJ, Brandsma CA, Heijink IH, Maksym GN, Walker D, Hackett TL.
Abstract
RATIONALE:Histological stains have been used as the gold standard to visualize extracellular matrix (ECM) changes associated with airwayremodeling in asthma, yet they provide no information on the biochemical and structural characteristics of the ECM, which are vital to understanding alterations in tissue function.
OBJECTIVE:Demonstrate the use of non-linear optical microscopy (NLOM) and texture analysis algorithms to image fibrillary collagen (Second harmonic generation) and elastin (Two-photon excited autofluorescence), to obtain biochemical and structural information on the remodeled ECM environment in asthma.
METHODS:Non-transplantable donor lungs from asthmatic (n=13) and control (n=12) donors, were used for the assessment of airway collagen and elastin fibers by NLOM, and extraction of lung fibroblasts for in vitro experiments.
MEASUREMENTS AND MAIN RESULTS:Fibrillar collagen is not only increased, but highly disorganized and fragmented within large and small asthmatic airways compared to controls, using NLOM imaging. Further, such structural alterations are present in pediatric and adult asthmatic donors, irrespective of fatal disease. In vitro studies demonstrated that asthmatic airway fibroblasts are deficient in their packaging of fibrillarcollagen-I, and express less decorin, important for collagen fibril packaging. Packaging of collagen fibrils was found to be more disorganized in asthmatics airways compared to controls, using transmission electron microscopy.
CONCLUSIONS:NLOM imaging enabled the structural assessment of the ECM, and the data suggest that airway remodeling in asthma involves the progressive accumulation of disorganized fibrillary collagen by airway fibroblasts. This study highlights the future potential clinical application of NLOM to assess airway remodeling in vivo.
基本原理：组织学染色已被用作可视化与哮喘气道重塑相关的细胞外基质（ECM）变化的金标准，但它们没有提供关于ECM的生化和结构特征的信息，而这对于理解组织功能的改变是至关重要的。
目的：展示使用非线性光学显微镜（NLOM）和纹理分析算法来成像原纤维胶原（二次谐波产生）和弹性蛋白（双光子激发自发荧光），以获得关于哮喘重塑ECM环境的生化和结构信息。
方法：来自哮喘（n = 13）和对照（n = 12）供体的非移植供体肺，通过NLOM评估气道胶原和弹性蛋白纤维，以及体外实验的肺成纤维细胞的提取。
检测和主要结果：与对照组相比，在哮喘组大、小气道内，使用NLOM成像的纤维状胶原蛋白不仅增加，而且高度混乱、碎裂。此外，这种结构改变存在于儿科和成人哮喘供体中，与致命疾病无关。体外研究表明，哮喘气道成纤维细胞对纤维胶原蛋白-I的组装有缺陷，对胶原纤维组装很重要的核心蛋白聚糖表达较少。使用透射电子显微镜观察发现，与对照相比，在哮喘气道中胶原原纤维的组装更加混乱。
结论：NLOM成像使得ECM的结构评估成为可能，并且数据表明哮喘中的气道重塑包括气道成纤维细胞引起的紊乱纤维胶原的不断积聚。该研究强调了NLOM用来评估体内气道重塑的潜在临床应用价值。
(中日友好医院呼吸与危重症医学科 顾宪民 摘译 林江涛 审校)
(Am J Respir Crit Care Med. 2019 Apr 5. doi: 10.1164/rccm. )
(Am J Respir Crit Care Med. 2019 Apr 5. doi: 10.1164/rccm. )
Defective Fibrillar Collagen Organization by Fibroblasts Contributes to Airway Remodeling in Asthma.
Mostaço-Guidolin LB, Osei ET, Ullah J, Hajimohammadi S, Fouadi M, Li X, Li V, Shaheen F, Yang CX, Chu F, Cole DJ, Brandsma CA, Heijink IH, Maksym GN, Walker D, Hackett TL.
Abstract
RATIONALE:Histological stains have been used as the gold standard to visualize extracellular matrix (ECM) changes associated with airwayremodeling in asthma, yet they provide no information on the biochemical and structural characteristics of the ECM, which are vital to understanding alterations in tissue function.
OBJECTIVE:Demonstrate the use of non-linear optical microscopy (NLOM) and texture analysis algorithms to image fibrillary collagen (Second harmonic generation) and elastin (Two-photon excited autofluorescence), to obtain biochemical and structural information on the remodeled ECM environment in asthma.
METHODS:Non-transplantable donor lungs from asthmatic (n=13) and control (n=12) donors, were used for the assessment of airway collagen and elastin fibers by NLOM, and extraction of lung fibroblasts for in vitro experiments.
MEASUREMENTS AND MAIN RESULTS:Fibrillar collagen is not only increased, but highly disorganized and fragmented within large and small asthmatic airways compared to controls, using NLOM imaging. Further, such structural alterations are present in pediatric and adult asthmatic donors, irrespective of fatal disease. In vitro studies demonstrated that asthmatic airway fibroblasts are deficient in their packaging of fibrillarcollagen-I, and express less decorin, important for collagen fibril packaging. Packaging of collagen fibrils was found to be more disorganized in asthmatics airways compared to controls, using transmission electron microscopy.
CONCLUSIONS:NLOM imaging enabled the structural assessment of the ECM, and the data suggest that airway remodeling in asthma involves the progressive accumulation of disorganized fibrillary collagen by airway fibroblasts. This study highlights the future potential clinical application of NLOM to assess airway remodeling in vivo.
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