摘要
目的:基于PI3K/AKT信号通路观察苦参碱对呼吸道合胞病毒(RSV)感染小鼠的保护作用机制.方法:采用RSV病毒悬液滴鼻感染C57BL/6J小鼠建立模型.实验分为3组:对照组、RSV组、RSV+苦参碱组,每组10只.苏木精—伊红(HE)染色及Masson染色法观察小鼠肺组织病理学变化,酶联免疫吸附试验(ELISA)法测定小鼠肺泡灌洗液炎症因子肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)和白介素-6(IL-6)水平,蛋白免疫印迹(western blotting)检测肺组织TNF-α、IL-1β、IL-6、Bax、Bcl-2、Caspase-3及PI3K/AKT信号通路关键蛋白的表达,免疫组化检测肺组织Bax、Caspase-3、PI3K、AKT表达.结果:与对照组比较,RSV组可见明显肺水肿,肺组织细胞呈现染色体聚集,核膜收缩,出现显著的肺部病变症状,同时,肺组织TNF-α、IL-1β、IL-6水平上升,Bax和Caspase-3表达水平升高,Bcl-2、P-PI3K、P-AKT表达水平降低(均P＜0.05).与RSV组相比,RSV+苦参碱组小鼠肺水肿显著缓解,肺部病变症状显著减轻,肺组织TNF-α、IL-1β、IL-6、Bax、Caspase-3表达水平显著降低,Bcl-2、P-PI3K、P-AKT表达显著升高(均P＜0.05).结论:苦参碱可抑制RSV导致的小鼠肺部炎症反应,促进肺组织细胞凋亡,其机制可能与激活PI3K/AKT信号通路有关.
Abstract
Objective:To explore the protective mechanism of matrine against respiratory syncytial virus(RSV)infection in mice based on the PI3K/AKT signaling pathway.Methods:The model of C57BL/6J mice infected with RSV virus by suspension nasal drops was established.The experiment was divided into three groups:control group,RSV group,RSV+matrine group,with 10 mice in each group.Pathological changes in mouse lung tissue were observed using hematoxylin-eosin staining(HE)and Masson staining.The levels of inflammatory factors tu-mor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in pulmonary alveolar lavage fluid of mice were determined by enzyme-linked immunosorbent assay(ELISA).Western blotting was performed to detect the expression of TNF-α,IL-1β,IL-6,Bax,Bcl-2,Caspase-3 and key proteins in the PI3K/AKT signaling pathway in lung tissue,and immunohistochemistry was used to detect the expression of Bax,Caspase-3,PI3K and AKT in lung tissue.Results:Compared with the control group,mice in the RSV group exhibited significant pulmonary edema,chromosomal aggregation in lung tissue cells,nuclear membrane pyknosis,marked symptoms of lung lesions,increased levels of TNF-α,IL-1β and IL-6 in lung tissue,increased expression levels of Bax and Caspase-3,and decreased expression levels of Bcl-2,P-PI3K and P-Akt(all P＜0.05).Compared with the RSV group,mice in the RSV+matrine group showed significantly alleviated pulmonary edema and pulmonary disease symptoms,significantly decreased expression levels of TNF-α,IL-1β,IL-6,Bax and Caspase-3 in lung tissue,and significantly increased expression levels of Bcl-2,P-PI3K and P-Akt(all P＜0.05).Conclusion:Matrine can inhibit the RSV-induced lung inflammation in mice and promote lung cell apoptosis,and its mechanism may be related to the activation of PI3K/AKT signaling pathway.
维普
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