Goldstein Benjamin D, Lauer Mark E, Caplan Arnold I, Bonfield Tracey L
Department of Pediatric Pulmonology, Rainbow Babies and Children's Hospital, University Hospitals Cleveland Medical Center, Cleveland, OH USA.
Cleveland Clinic Foundation, Department of Biomedical Engineering, Cleveland, OH USA.
J Inflamm (Lond). 2017 Aug 30;14:18. doi: 10.1186/s12950-017-0165-4. eCollection 2017.
Previous studies have demonstrated that ovalbumin sensitization promotes chronic asthma phenotype in murine asthma model. Human mesenchymal stem cells (hMSCs) are multipotent cells in vitro that have been shown to decrease inflammation and can reverse airway remodeling when infused into an in vivo chronic asthma model. However, the mechanism by which hMSCs reverse remodeling is still unclear. In this study, we hypothesized that hMSCs influence remodeling by decreasing extracellular matrix (ECM) deposition, more specifically by decreasing collagen I, collagen III, and hyaluronan synthesis.
Chronic asthma phenotype was produced in an in vitro model with 3 T3 murine airway fibroblast cells by stimulating with GM-CSF. Collagen I and collagen III gene expression was investigated using RT-PCR and Taqman techniques. Hyaluronan was evaluated using FACE and Western Blots. The chronic asthma phenotype was produced in vivo in murine model using sensitization with ovalbumin with and without hMSC infusion therapy. ECM deposition (specifically trichrome staining, soluble and insoluble collagen deposition, and hyaluronan production) was evaluated. Image quantification was used to monitor trichrome staining changes.
GM-CSF which induced collagen I and collagen III production was down-regulated with hMSC using co-culture. In the in vivo model, Ovalbumin induced enhanced ECM deposition, soluble and insoluble collagen production, and lung elastance. hMSC infusions decreased ECM deposition as evidenced by decreases in soluble and insoluble collagen production.
hMSCs participate in improved outcomes of remodeling by reversing excess collagen deposition and changing hyaluronan levels.
先前的研究表明，在小鼠哮喘模型中，卵清蛋白致敏可促进慢性哮喘表型。人间充质干细胞（hMSCs）是体外多能细胞，已显示其可减轻炎症，并且当注入体内慢性哮喘模型时可逆转气道重塑。然而，hMSCs逆转重塑的机制仍不清楚。在本研究中，我们假设hMSCs通过减少细胞外基质（ECM）沉积，更具体地说是通过减少I型胶原、III型胶原和透明质酸的合成来影响重塑。
在体外模型中，用GM-CSF刺激3T3小鼠气道成纤维细胞产生慢性哮喘表型。使用RT-PCR和Taqman技术研究I型胶原和III型胶原基因表达。使用FACE和蛋白质印迹法评估透明质酸。在体内小鼠模型中，通过卵清蛋白致敏并联合或不联合hMSC输注疗法产生慢性哮喘表型。评估ECM沉积（特别是三色染色、可溶性和不溶性胶原沉积以及透明质酸产生）。使用图像定量监测三色染色变化。
与hMSC共培养可下调诱导I型胶原和III型胶原产生的GM-CSF。在体内模型中，卵清蛋白诱导ECM沉积增加、可溶性和不溶性胶原产生增加以及肺弹性增加。hMSC输注减少了ECM沉积，可溶性和不溶性胶原产生减少证明了这一点。
hMSCs通过逆转过量的胶原沉积和改变透明质酸水平参与改善重塑结果。
