南五味子多糖对体外培养巨噬细胞吞噬功能与IL-12表达的影响  

Effects of Kadsura longepedunculata polysaccharide on phagocytosis and IL-12 expression of macrophages in vitro

作　　者：汪洋 张宏方[1] 于鹏龙[1] 李文侠 刘洋[1] 谭芷芬 WANG Yang;ZHANG Hongfang;YU Penglong;LI Wenxia;LIU Yang;TAN Zhifen(Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi,China)

机构地区：[1]陕西中医药大学,陕西咸阳712046

出　　处：《现代中西医结合杂志》2023年第6期770-775,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：陕西省科技厅自然基础科研基金项目(2019JM-551);陕西省中医药管理局科研项目(JCP002)。

摘　　要：目的研究南五味子多糖对巨噬细胞吞噬功能的影响及对巨噬细胞吞噬作用的免疫调节机制。方法依照中药多糖的提取方法从南五味子中提取粗多糖,从小鼠腹腔分离提取巨噬细胞并培养。采用CCK-8法考察50μg/mL、100μg/mL、200μg/mL、300μg/mL、400μg/mL、600μg/mL南五味子多糖对巨噬细胞增殖影响;设置空白组、南五味子多糖50μg/mL组、南五味子多糖150μg/mL组、南五味子多糖250μg/mL组,每组巨噬细胞给予相应干预48 h后,显微镜下观察活细胞的形态、墨汁和金黄色葡萄球菌吞噬情况,细胞酶化学染色观察巨噬细胞胞内酸性磷酸酶的阳性表达情况;设置空白组、南五味子多糖50μg/mL组、南五味子多糖100μg/mL组、南五味子多糖150μg/mL组,每组巨噬细胞给予相应干预48 h后,免疫组化法检测巨噬细胞中白细胞介素-12(IL-12)阳性表达情况。结果南五味子多糖浓度在50~200μg/mL时,巨噬细胞增殖活性递增性高于空白组(P均<0.05),但南五味子多糖浓度超过200μg/mL时,巨噬细胞的增殖活性逐渐减弱,浓度为600μg/mL时,巨噬细胞活性被抑制。与空白组比较,南五味子多糖50μg/mL组和南五味子多糖150μg/mL组巨噬细胞的体积明显增大,巨噬细胞吞噬墨汁阳性面积百分率、酸性磷酸酶阳性面积占比均显著增高(P均<0.05),吞噬金黄色葡萄球菌吸光度值均明显降低(P均<0.05);而南五味子多糖250μg/mL组巨噬细胞的体积形态无显著变化,巨噬细胞吞噬墨汁阳性面积百分率和吞噬金黄色葡萄球菌吸光度值、酸性磷酸酶阳性面积占比与空白组比较差异均无统计学意义(P均>0.05);南五味子多糖各组IL-12表达半定量面积均明显高于空白组(P均<0.05)。结论南五味子多糖在50~150μg/mL浓度范围内可激活巨噬细胞,增强巨噬细胞的吞噬功能和分泌IL-12的功能,发挥免疫促进作用,但浓度在250μg/mL时出现免疫抑制作用。Objective It is to study the effect of Kadsura longepedunculata polysaccharides(KLP)on phagocytic function of macrophages,and to explore the immunomodulatory mechanism on macrophage phagocytosis.Methods The crude polysaccharide was extracted from Kadsura longepedunculata according to the extraction method of traditional Chinese medicine polysaccharide,macrophages were isolated and culultured from mouse abdominal cavity.The effect of 50μg/mL,100μg/mL,200μg/mL,300μg/mL,400μg/mL,600μg/mL KLP on macrophage proliferation were detected by CCK-8 method.An blank group,50μg/mL KLP group,150μg/mL KLP group,250μg/mL KLP group were set up,after 48 hours of corresponding intervention on the macrophages in each group,the morphology of living cells,ink and Staphylococcus aureus phagocytized were observed under microscope,and the positive expression of intracellular acid phosphatase in macrophages was observed by cytochemical staining.An blank group,50μg/mL KLP group,100μg/mL KLP group,150μg/mL KLP group were set up,after 48 hours of corresponding intervention on the macrophages in each group,the positive expression of interleukin 12(IL-12)in macrophages was detected by immunohistochemical method.Results The increasing proliferative activity of macrophages was higher when the concentration of KLP was between 50-200μg/mL(P<0.05),but the proliferative activity of macrophages was gradually decreased when the concentration of KLP exceeded 200μg/mL,and the proliferative activity of macrophages was inhibited when the concentration was 600μg/mL.Compared with the blank group,the volume of macrophages in the 50μg/mL KLP group and 150μg/mL KLP group were significantly increased,with a significant increase in the percentage of ink positive areas and the proportion of acid phosphatase positive areas swallowed by macrophages(P<0.05),and a significant decrease in the absorbance value of Staphylococcus aureus swallowed(P<0.05);There was no significant change in the volume and morphology of macrophages in the 250μg/mL KLP grou

关 键 词：南五味子多糖 巨噬细胞 免疫调节作用 

分 类 号：R-332[医药卫生]