枸杞果多糖对哮喘小鼠巨噬细胞的免疫调节作用

Immunomodulatory Effects of Lycium Barbarum Polysaccharide on Macrophages in Asthmatic Mice

目的 探究枸杞果多糖(lycium barbarum polysaccharide,LBPs)对小鼠巨噬细胞的免疫调节作用。方法 以卵清蛋白(ovalbumin,OVA)诱导的哮喘小鼠为模型,并给与LBPs灌胃干预。将30只雌性BALB/C小鼠随机分为三组:PBS对照组、哮喘模型组、LBPs+哮喘模型组,每组10只。通过HE染色观察小鼠肺组织结构的病理学变化,使用流式细胞术检测小鼠肺组织中巨噬细胞的总数以及M1巨噬细胞的占比。以不同剂量的LBPs(50、100、150μg)干预100 ng·mL^(-1)脂多糖(lipopolysaccharide,LPS)诱导的RAW264.7小鼠巨噬细胞,分组为PBS对照组、M1模型组、50μg LBPs+M1模型组、100μg LBPs+M1模型组、150μg LBPs+M1模型组,观察LBPs对小鼠巨噬细胞形态变化的影响,流式细胞术检测各组中M1型巨噬细胞的占比。结果 HE染色表明LBPs可以减轻哮喘小鼠的炎症水平。与PBS对照组总巨噬细胞(4.43%)和M1巨噬细胞(3.75%)相比,哮喘模型组肺部总巨噬细胞和M1巨噬细胞的比例分别增加至6.99%和7.79%(P均<0.05);与哮喘模型组比较,经过LBPs干预后,小鼠肺部总巨噬细胞和M1巨噬细胞的比例分别下降至5.51%和4.36%(P均<0.05)。在RAW264.7小鼠巨噬细胞实验中,与PBS对照组M1型巨噬细胞(0.22%)相比,在LPS诱导的模型组中M1巨噬细胞的比例增加(P<0.05);与M1模型组(57.1%)比较,50、100、150μg的LBPs干预组,M1细胞的比例分别下降至52.3%(P>0.05)、51.4%和46.9%(P均<0.05)。结论 LBPs可以减少哮喘小鼠总巨噬细胞和M1型巨噬细胞的比例,LBPs可以减少LPS诱导的M1型RAW264.7巨噬细胞的占比。

Objective To investigate the immunological regulation of lycium barbarum polysaccharide(LBPs)on macrophages in mice.Methods Asthma mice induced by ovalbumin(OVA)were treated with LBPs.Thirty female BALB/C mice were randomly divided into three groups:PBS control group,asthma model group and LBPs+ asthma model group,with 10 mice in each group.HE staining was used to observe the pathological changes of lung tissue structure of mice.Flow cytometry was used to detect the total number of macrophages and the proportion of M1 macrophages in mouse lung tissue.LBPs at different doses(50 μg,100 μg,150 μg)were used to intervene 100 ng·mL^(-1) lipopolysaccharide(LPS)-induced RAW264.7 mouse macrophages,which were divided into:PBS control group,M1 model group,50 μg+M1 model group,100 μg+M1 model group and150 μg+M1 model group were used to observe the effects of LBPs on the morphologic changes of macrophages in mice,and the proportion of M1 macrophages in each group was detected by flow cytometry.Results HE staining showed that LBPs could reduce the level of inflammation in asthmatic mice.Compared with total macrophages(4.43%)and M1 macrophages(3.75%)in PBS control group,the proportion of total macrophages and M1 macrophages in asthma model group increased to 6.99% and 7.79%,respectively(P all<0.05).Compared with asthma model group,after LBPs intervention,the proportion of total macrophages and M1 macrophages in the lungs of mice decreased to 5.51% and 4.36%,respectively(P all<0.05).In RAW264.7 mouse macrophage level experiments,the proportion of M1 macrophages in the LPS-induced model group was increased compared with M1 macrophages in the PBS control group(0.22%)(P <0.05).Compared with the M1 model group(57.1%),the proportion of M1 cells in the 50 μg,100 μg and 150 μg LBPs intervention groups decreased to52.3%(P>0.05),51.4%,and 46.9%,respectively(P all <0.05).Conclusion LBPs reduced the proportion of total macrophages and M1 macrophages in asthmatic mice,and LBPs reduced the proportion of LPS-induced M1-type RAW264.7 macrophages.