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鹰嘴豆多肽的免疫活性研究
王亚非1, 于淼淼2, 于 悦,等1
1.吉林农业大学 食品科学与工程学院;2.长春海关技术中心
摘要:
【目的】探究鹰嘴豆多肽对机体免疫功能的调节作用。【方法】用复合酶解方法制备鹰嘴豆多肽,对其分子质量分布及氨基酸组成进行分析。给小鼠腹腔注射环磷酰胺建立免疫功能低下的动物模型,建模成功后将小鼠随机均分为模型组(灌胃生理盐水)、阳性对照组(灌胃匹多莫德25 mg/kg)和鹰嘴豆多肽低、中、高剂量组(分别灌胃鹰嘴豆多肽50,100和200 mg/kg),另取10只健康小鼠作为空白组(灌胃生理盐水)。各组小鼠每日灌胃1次相应物质,连续灌胃21 d后采样,研究鹰嘴豆多肽对小鼠体质量、脾脏和胸腺指数及形态变化、脾淋巴细胞增殖率、血清细胞因子(IFN-γ、IL-6、IL-2)及免疫球蛋白(IgG、IgA和IgM)质量浓度的影响。研究不同质量浓度(50,100,200和400 μg/mL)鹰嘴豆多肽对RAW264.7细胞功能的免疫调节作用,以MTT法、中性红法和酶联免疫吸附检测试剂盒来检测细胞增殖率、吞噬活性以及分泌的细胞因子(NO、TNF-α、IL-6、IL-1β)水平。【结果】制得分子质量在1 000 u左右、纯度为97.12 %的鹰嘴豆多肽,其中富含谷氨酸、天冬氨酸、精氨酸、赖氨酸、亮氨酸、丝氨酸等氨基酸。与空白组相比,模型组小鼠脾脏和胸腺受损严重,多项检测指标显著降低;而鹰嘴豆多肽各剂量组小鼠的各项检测指标均较模型组小鼠显著增加,且呈剂量效应关系。鹰嘴豆多肽可促进RAW264.7细胞的增殖率和吞噬能力,并提升其产生的NO、IL-6、IL-1β和TNF-α的水平。【结论】鹰嘴豆多肽可以显著改善环磷酰胺导致的小鼠多项免疫相关指标的下降情况,同时可刺激巨噬细胞,增强其吞噬能力,促进其分泌白介素等细胞因子,表明鹰嘴豆多肽具有良好的免疫增强作用。
关键词:  鹰嘴豆多肽  免疫活性调节  RAW264.7细胞  细胞因子
DOI:
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基金项目:吉林省教育厅项目(JKH20210342KJ)
Immunological activity of chickpea polypeptides
WANG Yafei,YU Miaomiao,YU Yue,et al
Abstract:
【Objective】This study explored the regulatory effect of chickpea peptides on immune activity.【Method】First,cyclophosphamide was used to establish a mouse model of immunocompromised function,and the effects of low (50 mg/kg),medium (100 mg/kg) and high (200 mg/kg) doses of chickpea peptides on mouse body weight,spleen and thymus index and their morphology,splenic lymphocyte prolife-ration rate,serum cytokines (IFN-γ,IL-6 and IL-2) and immunoglobulin (IgG,IgA and IgM) levels were studied.Secondly,the effects of chickpea peptides (50,100,200 and 400 μg/mL) on immunomodulatory function of RAW264.7 cells were explored with MTT method,neutral red method and enzyme linked immunosorbent assay kit by detecting cell proliferation rate,phagocytic activity,and secreted cytokine (NO,TNF-α,IL-6 and IL-1β) levels.【Result】The prepared chickpea polypeptides had a molecular weight of about 1 000 u and a purity of 97.12% and were rich in amino acids such as glutamic acid,aspartic acid,arginine,lysine,leucine,and serine.Compared with the blank control group,the spleen and thymus of mice in the model group were severely damaged,and multiple detection indicators were significantly reduced.The detection indicators of mice in the chickpea polypeptide groups were significantly higher than those of the model group with a dose effect relationship.Chickpea peptides promoted the proliferation rate and phagocytosis of RAW264.7 cells,and increased the levels of produced NO,IL-6,IL-1β,and TNF-α.【Conclusion】Chickpea polypeptides can significantly improve the declines of multiple immune related indicators in mice caused by cyclophosphamide,and at the same time it stimulated macrophages,enhanced their phagocytic ability,and promoted the secretion of interleukins and other cytokines.It is concluded that chickpeas polypeptides have good immune enhancement effects.
Key words:  chickpea peptides  immune activity regulation  RAW264.7 cells  cytokines