水飞蓟宾对哮喘幼鼠肺内炎症反应的影响及可能机制

Effects of silybin on lung inflammation in asthmatic young mice and its possible mechanism

目的 探讨水飞蓟宾对哮喘幼鼠肺内炎症反应的影响及可能机制。方法 45只SPF级BALB/c幼鼠随机分为对照组(Control)、哮喘组(Model)和水飞蓟宾组(Silybin),每组15只,采用卵清蛋白致敏(OVA)制备幼鼠支气管哮喘模型。收集各组小鼠支气管肺泡灌洗液(BALF),进行总细胞计数和白细胞分类计数,ELISA方法检测各组小鼠BALF中白介素-1β(IL-1β)、白介素-4(IL-4)和肿瘤坏死因子α(TNF-α)的表达水平;HE染色观察各组小鼠肺组织病理学变化;免疫组织化学方法与Western blot方法检测各组小鼠肺组织TLR4与NF-κB p65蛋白的表达。结果 与哮喘组相比,水飞蓟宾组幼鼠BALF中细胞总数及嗜酸性粒细胞数明显降低(P<0.01),BALF中炎性因子IL-1β、IL-4与TNF-α的表达水平明显降低(P<0.01)。HE染色结果显示,水飞蓟宾能够减轻哮喘幼鼠肺组织的炎症反应。免疫组织化学方法与Western blot结果显示,水飞蓟宾能够减轻哮喘幼鼠肺组织TLR4与NF-κB p65蛋白的表达(P<0.01)。结论 水飞蓟宾能够通过抑制TLR4/NF-κB信号通路抑制哮喘幼鼠的肺内炎性反应。

Objective To investigate the effect of silybin on lung inflammation in asthmatic mice and its possible mechanism. Methods Forty-five SPF BALB/C young mice were randomly divided into control group, asthma group and silybin group, with 15 young mice in each group. The young mice were sensitized by ovalbumin(OVA) to establish bronchial asthma model. Bronchoalveolar lavage fluid(BALF) of mice in each group was collected, and total cell count and leukocyte classification count were performed. The levels of interleukin-1 β(IL-1β), interleukin-4(IL-4) and tumor necrosis factor α(TNF-α) in BALF of mice in each group were detected by ELISA. HE staining was used to observe the pathological changes of lung tissues in each group. The expression of TLR4 and NF-κB P65 protein in lung tissues of each group was detected by immunohistochemistry and Western blot. Results Compared with asthma group, the total number of cells and eosinophils in BALF were significantly decreased in silybin group(P<0.01), and the levels of IL-1β, IL-4 and TNF-α in BALF were significantly decreased in silybin group(P<0.01). The results of HE staining showed that silybin could reduce the inflammatory response of lung tissue in asthmatic young mice. The results of immunohistochemical and Western blot showed that silybin could reduce the expression of TLR4 and NF-κB P65 protein in lung tissues of asthmatic young mice(P<0.01). Conclusion Silybin can inhibit pulmonary inflammatory response in asthmatic young mice by inhibiting TLR4/NF-κB signaling pathway.