猪皮胶原肽对RAW264.7细胞的免疫调节作用

Immunomodulatory effects of porcine skin collagen peptides on RAW264.7 cells

摘要

为探究猪皮胶原肽对RAW264.7细胞的免疫调节作用,本研究通过酶解制备猪皮胶原肽,并对其氨基酸组成及分子量分布进行分析。采用MTT法、中性红吞噬实验、细胞形态学观察、细胞因子含量测定以及实时荧光定量PCR等方法,系统评估猪皮胶原肽对RAW264.7细胞增殖活性、吞噬功能、细胞形态变化、NO、TNF-α、IL-2和IL-12生成水平以及相关免疫因子基因(iNOS、TNF-α、IL-2和IL-12) mRNA表达的影响。结果表明,猪皮胶原肽主要由甘氨酸、脯氨酸、丙氨酸、谷氨酸、天冬氨酸和精氨酸组成,分子量主要在500~5000 Da之间,可显著促进RAW264.7细胞的增殖和吞噬能力,改变细胞形态,促进NO、TNF-α、IL-2和IL-12的生成,并上调iNOS、TNF-α、IL-2和IL-12的基因表达。综上所述,猪皮胶原肽具有激活巨噬细胞、增强机体非特异性免疫功能的潜力。

Abstract

To investigate the immunomodulatory effects of porcine skin collagen peptides on RAW264.7 cells, this study prepared Porcine skin collagen peptides via enzymatic hydrolysis and analyzed their amino acid composition and molecular weight distribution. The effects of porcine collagen peptides on RAW264.7 cells were systematically evaluated using a range of methods, including the MTT assay, neutral red phagocytosis assay, cell morphological observation, cytokine content measurement, and real-time quantitative PCR. These analyses assessed cell proliferation activity, phagocytic function, morphological changes, the production levels of NO, TNF-α, IL-2, and IL-12, as well as the mRNA expression of related immune factors (iNOS, TNF-α, IL-2, and IL-12). The results demonstrated that Porcine skin collagen peptides are primarily composed of glycine, proline, alanine, glutamic acid, aspartic acid, and arginine, with molecular weights mainly distributed between 500 and 5000 Da. Porcine skin collagen peptides significantly promoted RAW264.7 cell proliferation and phagocytosis, induced morphological changes, enhanced NO, TNF-α, IL-2, and IL-12 production, and upregulated the gene expression of iNOS, TNF-α, IL-2, and IL-12. In conclusion, Porcine skin collagen peptides exhibit potential in activating macrophages and enhancing non-specific immune functions.

关键词

猪皮胶原肽 / RAW264.7细胞 / 酶解 / 免疫调节作用 / 氨基酸组成

Key words

porcine skin collagen peptides / RAW264.7 cells / enzymatic hydrolysis / immunomodulatory effects / amino acid composition

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基金资助

山东省科技型中小企业创新能力提升工程项目(2023TSGC0167)

山东省科技型中小企业创新能力提升工程项目(2023TSGC0169)

国家重点研发计划政府间国际科技创新合作项目(2023YFE0111900)

烟台开发区科技领军人才项目(2020CXRC4)

山东省重点研发计划项目(2023LZGC005)

山东省重点研发计划项目(2023CXPT037)

泉城产业领军人才支持计划(2092021033)

“海右人才”产业领军人才支持项目(1362022058)