■穿心莲内酯磺酸盐(Andro-S),中药,在中国通常用于治疗小儿呼吸道感染。然而,其在呼吸道合胞病毒(RSV)引起的感染中的治疗作用尚未报道。因此,我们旨在使用RSV感染诱导的气道炎症的小鼠模型研究Andro-S的治疗效果。
■用RSV鼻内感染免疫功能低下(环磷酰胺治疗)的BALB/c小鼠,用鼻内或腹膜内Andro-S治疗,每天一次,连续五天,从感染的那一天开始。使用苏木精和伊红染色评估肺的组织病理学变化。总炎性细胞计数和巨噬细胞,淋巴细胞,中性粒细胞,显微镜下测定支气管肺泡灌洗液(BALF)中的嗜酸性粒细胞计数。使用酶联免疫吸附测定(ELISA)检测BALF中的干扰素-γ(IFN-γ)水平。采用实时定量聚合酶链反应(qRT-PCR)检测肺组织中RSV核蛋白(N)和Toll样受体(TLRs)1-9的信使RNA水平。RSVN的蛋白质水平,RSV融合蛋白(F),通过蛋白质印迹分析检测TLR2、TLR3和含有TIR结构域的衔接子诱导干扰素-β(TRIF)。
■RSV感染引起肺部炎症,表现为毛细支气管炎,肺泡炎,和血管周围炎症;增加炎症细胞的数量;和BALF中IFN-γ水平升高。感染小鼠肺部炎症与肺部RSVN水平呈正相关。鼻内Andro-S显著下调RSVN,RSVF,TLR3和TRIF蛋白在肺部表达并改善感染动物的肺部炎症。然而,腹膜内Andro-S对RSV感染引起的肺部炎症无影响。
■鼻内Andro-S通过下调TLR3和TRIF抑制RSV复制并改善RSV感染诱导的肺部炎症。因此,鼻内施用可以是用于治疗RSV感染的合适的药物递送方法。
UNASSIGNED: Andrographolide sulfonate (Andro-S), a traditional Chinese medicine, is commonly used to treat pediatric respiratory tract infections in China. However, its therapeutic effects in infections caused by respiratory syncytial virus (RSV) have not been reported. We thus aimed to investigate the therapeutic effects of Andro-S using a mouse model of RSV infection-induced airway inflammation.
UNASSIGNED: Immunocompromised (cyclophosphamide-treated) BALB/c mice were intranasally infected with RSV and treated with intranasal or intraperitoneal Andro-S once daily for five consecutive days, starting on the day of infection. Histopathological changes in the lung were evaluated using hematoxylin and eosin staining. Total inflammatory cell counts and macrophage, lymphocyte, neutrophil, and eosinophil counts in the bronchoalveolar lavage fluid (BALF) were microscopically determined. Interferon-γ (IFN-γ) levels in the BALF were detected using enzyme-linked immunosorbent assay (ELISA). The messenger RNA levels of RSV nucleoprotein (N) and Toll-like receptors (TLRs) 1-9 in lung tissues were determined with quantitative real-time polymerase chain reaction (qRT-PCR). The protein levels of RSV N, RSV fusion protein (F), TLR2, TLR3, and TIR domain-containing adapter-inducing interferon-β (TRIF) were detected via Western blot analysis.
UNASSIGNED: RSV infection caused lung inflammation, manifesting as bronchiolitis, alveolitis, and perivascular inflammation; increased the number of inflammatory cells; and elevated IFN-γ levels in the BALF. Lung inflammation was positively correlated with pulmonary RSV N levels in infected mice. Intranasal Andro-S significantly downregulated RSV N, RSV F, TLR3, and TRIF protein expression in the lung and ameliorated lung inflammation in infected animals. However, intraperitoneal Andro-S showed no effects on lung inflammation caused by RSV infection.
UNASSIGNED: Intranasal Andro-S inhibits RSV replication and ameliorates RSV infection-induced lung inflammation by downregulating TLR3 and TRIF. Therefore, intranasal administration may be a suitable drug delivery method for treating RSV infection.