鹿胎肽对巨噬细胞RAW264.7的免疫调节作用

Immunomodulatory Effects of Peptides from Deer Fetus on RAW264.7 Macrophages

本实验从鹿胎中提取鹿胎肽(DFP),通过体外实验研究其对巨噬细胞RAW264.7的免疫调节作用。采用噻唑蓝(MTT)实验检测超滤组分各肽段的增殖活性,筛选增殖活性高的组分。选用中性红吞噬实验、酶联免疫吸附检测法测定DFP对RAW264.7细胞吞噬指数、一氧化氮(NO)浓度及细胞因子如白介素-1β(interleukin-1β,IL-1β)、IL-6和肿瘤坏死因子子-α(tumor necrosis factor-α,TNF-α)的分泌能力,同时还研究了DFP对巨噬细胞各周期分布的影响。结果表明,分子质量小于1 kDa的DFP-5对巨噬细胞RAW264.7作用后,可显著(P<0.05)提高巨噬细胞的增殖活性、吞噬活性、NO释放量以及细胞因子的分泌水平,并且在DFP-5质量浓度为25μg/mL时效果最好。细胞周期结果表明,DFP-5能够促进RAW264.7细胞的生长,且主要作用在G0/G1期。以上结果表明,鹿胎肽具有增强免疫能力的潜在作用。

Deer fetus peptides(DFP)was extracted from deer fetus and its immunomodulatory activities in macrophage RAW264.7 was evaluated in vitro.The proliferative activity of each peptide segment in ultrafiltration fraction was determined by methyl thiazolyltetrazolium(MTT)assay,and the components with high proliferative activity were screened.To further evaluate its immunomodulatory activity,the effects of DFP on the phagocytic activity,nitric oxide(NO)and cytokines secretion of RAW264.7 macrophages were detected by inverted microscope observation,neutral red phagocytosis assay and enzyme-linked immunosorbent assay(ELISA),respectively.The effects on the cell cycle distribution of macrophages were also observed.The results showed that DFP-5,with molecula mass less than 1 kDa,could significantly(P<0.05)increase cytokines in RAW264.7 such as nitric oxide(NO),interleukin-1β(IL-1β),interleukin-6(IL-6)and the tumor necrosis factor-α(TNF-α).When the concentration of DFP was 25μg/mL,the secretion of these factors was the closest to that of the positive control group(LPS).Cell cycle results showed that DFP could promote the proliferation of RAW264.7 cells and the effect was mainly at the G0/G1 phase.All these results indicated that the deer fetus peptide had the potential to activate macrophages and enhance immune function.