Abstract：
Persistent type (T) 2 airway inflammation plays an important role in the development of severe asthma. However, the molecular mechanisms leading to T2 severe asthma have yet to be fully clarified. Human normal lung epithelial cells (BEAS-2B cells) were transfected with LINC00158/BCL11B plasmid/small interfering RNA (siRNA). Levels of epithelial-mesenchymal transition (EMT)-related markers were measured using real-time qPCR (RT-qPCR) and western blot. A dual luciferase reporter assay was used to validate the targeting relationship between LINC00158 and BCL11B. The effects of LINC00158-lentivirus vector-mediated overexpression and dexamethasone on ovalbumin (OVA)/lipopolysaccharide (LPS)-induced severe asthma were investigated in mice in vivo. Our study showed that overexpression of LINC00158/BCL11B inhibited the levels of EMT-related proteins, apoptosis, and promoted the proliferation of BEAS-2B cells. BCL11B was a direct target of LINC00158. And LINC00158 targeted BCL11B to regulate EMT, apoptosis, and cell proliferation of BEAS-2B cells. Compared with severe asthma mice, LINC00158 overexpression alleviated OVA/LPS-induced airway hyperresponsiveness and airway inflammation, including reductions in T helper 2 cells factors in lung tissue and BALF, serum total- and OVA-specific IgE, inflammatory cell infiltration, and goblet cells hyperplasia. In addition, LINC00158 overexpression alleviated airway remodeling, including reduced plasma TGF-β1 and collagen fiber deposition, as well as suppression of EMT. Additionally, overexpression of LINC00158 enhanced the therapeutic effect of dexamethasone in severe asthmatic mice models. LINC00158 regulates BEAS-2B cell biological function by targeting BCL11B. LINC00158 ameliorates T2 severe asthma in vivo and provides new insights into the clinical treatment of severe asthma.
摘要：
持续性(T)2型气道炎症在重症哮喘的发生发展中起重要作用。然而,导致T2重度哮喘的分子机制尚未完全阐明.用LINC00158/BCL11B质粒/小干扰RNA(siRNA)转染人正常肺上皮细胞(BEAS-2B细胞)。使用实时qPCR(RT-qPCR)和蛋白质印迹测量上皮-间质转化(EMT)相关标志物的水平。使用双荧光素酶报告基因测定来验证LINC00158和BCL11B之间的靶向关系。在小鼠体内研究了LINC00158-慢病毒载体介导的过表达和地塞米松对卵清蛋白（OVA）/脂多糖（LPS）诱导的严重哮喘的影响。我们的研究表明，LINC00158/BCL11B的过表达抑制了EMT相关蛋白的水平，凋亡，并促进BEAS-2B细胞的增殖。BCL11B是LINC00158的直接靶标。LINC00158针对BCL11B来监管EMT，凋亡，和BEAS-2B细胞的细胞增殖。与重度哮喘小鼠相比，LINC00158过表达减轻OVA/LPS诱导的气道高反应性和气道炎症,包括肺组织和BALF中辅助性T细胞2因子的减少，血清总特异性和OVA特异性IgE，炎性细胞浸润，和杯状细胞增生。此外,LINC00158过表达减轻气道重塑，包括降低血浆TGF-β1和胶原纤维沉积，以及EMT的抑制。此外,LINC00158的过度表达增强了地塞米松在重度哮喘小鼠模型中的治疗效果。LINC00158通过靶向BCL11B调节BEAS-2B细胞生物学功能。LINC00158可在体内改善T2重症哮喘，为重症哮喘的临床治疗提供新的见解。
