羊骨胶原肽对大鼠腹腔巨噬细胞免疫能力的影响  

Effect of sheep bone collagen peptide on immune function in rat peritoneal macrophages

作　　者：张慧琴 霍乃蕊[1] 冀霞 段卓 马林峰 古少鹏[1] ZHANG Huiqin;HUO Nairui;JI Xia;DUAN Zhuo;MA Linfeng;GU Shaopeng(College of Veterinary Medicine,Shanxi Agricultural University,Jinzhong 030801,China)

机构地区：[1]山西农业大学动物医学学院,山西晋中030801

出　　处：《中国实验动物学报》2021年第2期176-182,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(31201347);山西省重点研发计划(201803D221024-2)。

摘　　要：目的探究羊骨胶原肽(sheep bone collagen peptide,SBCP)对正常状态及脂多糖(lipopolysaccharide,LPS)活化状态巨噬细胞免疫能力的影响。方法以不同剂量的SBCP作用于未经LPS活化和LPS活化的大鼠腹腔巨噬细胞,分别以MTT法和中性红法测定巨噬细胞代谢活力和吞噬活性,Griess法测定NO分泌量,ELISA检测TNF-α、IL-6含量,qRT-PCR测定Toll样受体(Toll-like receptors,TLRs)通路相关基因及负性调控因子单Ig区IL-1相关受体(Single Ig IL-1-related receptor,SIGIRR)的mRNA表达量。结果SBCP作用于正常状态的巨噬细胞,可提高其代谢活力,吞噬活性及NO、TNF-α、IL-6分泌量,上调TLRs通路相关基因表达量,下调SIGIRR mRNA表达量,均呈剂量依赖性,SBCP剂量为105μg/mL时,与LPS空白组无显著性差异。SBCP作用于LPS活化的巨噬细胞时,可抑制其代谢活力,以102μg/mL效果最佳;对吞噬活性及NO、TNF-α、IL-6分泌量的抑制均以103μg/mL最佳;并下调TLRs相关基因表达量且上调SIGIRR mRNA的表达量。结论SBCP通过TLRs信号通路对炎症反应发挥双向调节作用,剂量为10~104μg/mL时可增强巨噬细胞的免疫能力,105μg/mL时可诱导巨噬细胞释放大量炎症介质。而对LPS诱导的炎症反应,SBCP各剂量组均具有抑制作用。Objective To investigate the effect of sheep bone collagen peptide(SBCP)on the immune function of macrophages with or without stimulation with lipopolysaccharide(LPS).Methods Rat peritoneal macrophages were treated with different doses of SBCP,with or without stimulation with LPS.The metabolic activity and phagocytic ability of macrophages were measured by MTT assay and the neutral red method,respectively.The production of NO was measured with the Griess reaction,and TNF-α and IL-6 were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA expression levels of key genes in the Toll-like receptor pathway and the negative regulatory factor Single Ig IL-1-related receptor were measured by qRT-PCR.Results SBCP treatment of normal macrophages increased their metabolic activity,phagocytosis,NO,TNF-α and IL-6 production,and upregulated mRNA expression of key genes in the TLR pathway and downregulated SIGIRR mRNA expression in a dose-dependent manner.At a concentration of 105μg/mL,there was no significant difference between the SBCP and LPS group.SBCP treatment of LPS-stimulated macrophages inhibited their metabolic activity,with 102μg/mL having the greatest effect,while 103μg/mL most strongly inhibited phagocytosis and the secretion of NO,TNF-α and IL-6.This concentration also downregulated the mRNA expression of TLR-related genes and upregulated the mRNA expression of SIGIRR.Conclusions SBCP has a dual regulatory effect on the inflammatory response through the TLR signaling pathway.At a concentration of 10~104μg/mL,it enhances the immune activity of macrophages,while at 105μg/mL,it induces macrophages to release various inflammatory mediators.However,SBCP inhibits the inflammatory reaction induced by LPS.

关 键 词：羊骨胶原肽 巨噬细胞 免疫功能 Toll样受体(TLRs) 炎症因子 

分 类 号：Q95-33[生物学—动物学]