基础医学与临床 ›› 2016, Vol. 36 ›› Issue (10): 1417-1421.

• 研究论文 • 上一篇    下一篇

栀子苷减轻幼年哮喘小鼠气道炎性反应并下调TLR4/NF-κB活性

刘雨东,王美霞,邓跃林,罗建峰,柴华,李杨柳,孙新   

  1. 陕西省西安市西京医院
  • 收稿日期:2015-11-12 修回日期:2016-03-28 出版日期:2016-10-05 发布日期:2016-09-27
  • 通讯作者: 孙新 E-mail:boyue145@126.com

Geniposide attenuates airway inflammation and down-regulation TLR4/NF-κBactivity in infant asthma mice

  • Received:2015-11-12 Revised:2016-03-28 Online:2016-10-05 Published:2016-09-27

摘要: 目的 探讨栀子苷(Gen)对幼年哮喘小鼠气道炎症和平滑肌细胞(ASMCs)损伤的保护作用。方法 将小鼠分为对照组、哮喘组(Ova)、Gen组(Ova+Gen,80mg/kg)(每组n=10)。苏木精-伊红(H&E)和阿辛兰-过碘酸雪夫(AB-PAS)染色观察肺组织病理变化(细胞浸润和杯状黏液细胞分泌);观察支气管肺泡灌洗液(BALF)中炎性反应细胞嗜酸性粒细胞、中性粒细胞和巨噬细胞数目;ELISA检测BALF中TLR4及Th2型细胞因子IL-4,IL-5及IL-13表达。体外分离和培养ASMCs,分为对照组、哮喘组(Ova)、低剂量Gen组(20mg/L)、中剂量Gen组(50mg/L)和高剂量Gen组(100mg/L)(每组n=8)。qRT-PCR和Western blot检测TLR4和NF-κB p65表达;四甲基偶氮唑蓝法(MTT)检测ASMCs增殖。结果 体内实验发现,Gen可显著降低肺部细胞浸润和杯状黏液细胞分泌;减少炎性反应细胞增多(P<0.01);降低TLR4和Th2型细胞因子表达(P<0.01)。体外实验发现,Gen可抑制TLR4/NF-κB表达;抑制ASMCs增殖。结论 Gen可能通过抑制TLR4/NF-κB活化减轻幼年哮喘小鼠气道炎性反应和SMCs损伤。

关键词: 栀子苷, 哮喘, 炎症, 平滑肌细胞, TLR4/NF-κB

Abstract: Objective To explore the protective effects of geniposide on airway inflammation and smooth muscle cells (ASMCs) injury in infant asthma mice. Methods BALA/c mice were divided into three groups,including control group, asthmagroup, and geniposide group(80 mg/kg) (10 in each group). The pathological changes in experimental lung tissues, including cells infiltrating, goblet mucus secretion were detected by H&E staining and AB-PAS staining.The number of inflammatory cells in bronchoalveolar lavage fluid (BALF), including Eosinophils, Neutrophils, and Macrophages, was also observed.The levels of TLR4 and Th2 type cytokines IL-4, IL-5 and IL-13 in the BALF were determined by ELISA assay. Then, ASMCs were isolated and cultured in vitro. Cultured cells were divided into five groups, control group, asthma group (Ova), low dose of geniposide group(20mg/L), medium dose of geniposide group (50mg/L) and high dose of geniposide group (100mg/L)(8 in each group). The expression of TLR4 and NF-κB p65 were detected by qRT-PCR and Western blot analyses.The cell proliferation ability of ASMCs was determined using the MTT assay. Results In vivo studies, the results indicated that geniposide induced the decrease of asthma lung cells infiltrating and goblet mucus cells secretion, and the increase of inflammation cells(P<0.01),as well as the reduction of TLR4 and Th2 type cytokines secretion(P<0.01).In vitro studies, the results demonstrated that geniposide inhibited TLR4/NF-κB activation, and restrained cell proliferation in asthma-derived ASMCs. Conclusions The effect of Geniposide reduces airway inflammation and ASMCs injury in infant asthma mice, which may be through the ability of inhibiting the TLR4/NF-κB activation.