蔓菁粗多糖对RAW264.7巨噬细胞的免疫活性影响研究  

Study on the Immune Activity of RAW264.7 Macrophages Affected by Crude Polysaccharides from Manjing

作　　者：牛丹 赵静宇 王喜明 薛莉 李卫红 NIU Dan;ZHAO Jingyu;WANG Ximing;XUE Li;LI Weihong(Medicines Evaluation Center of Shanxi Province,Shanxi Institllte of Medicine and Life Sciences,Taiyuan,Shanxi 030006,China;Center for Drug Inspection of Shanxi Province,Center for Vaccine Inspection of Shanxi Province,Taiyuan,Shanxi 030032,China)

机构地区：[1]山西省药品审评中心,山西省医药与生命科学研究院,山西太原030006 [2]山西省药品检查中心,山西省疫苗检查中心,山西太原030032

出　　处：《农产品加工》2024年第23期6-11,共6页Farm Products Processing

基　　金：山西省科技厅面上自然基金项目“山西特色农产品——蔓菁免疫调节活性成分研究”(201801D121296)。

摘　　要：通过体外细胞试验,探讨蔓菁粗多糖对RAW264.7巨噬细胞的免疫活性影响。取对数生长期的巨噬细胞,调整细胞含量至1.2×10^(5)个/mL,接种于96孔板培养24 h,分别设置阴性对照(只加完全培养基),阳性对照组(LPS,1μg/mL),蔓菁粗多糖组(50,100,200,400μg/mL),培养24 h后MTT法检测细胞增殖活力、吞噬能力、分泌NO能力;小鼠颈椎脱臼处死,无菌环境下取脾脏,调整细胞含量为1.0×10^(6)个/mL,接种于96孔板,设对照组、蔓菁粗多糖组(50,100,200,400μg/mL)以及Con A(10μg/mL)阳性对照组或LPS(5μg/mL)阳性对照组,培养24,48,72 h。培养结束前4 h,每孔加入MTT(5 mg/mL)10μL,离心弃上清液,加入100μL DMSO充分振荡后,用酶标仪测定波长570 nm处的吸光度,计算增殖指数。与阴性对照组相比,蔓菁粗多糖组(50μg/mL)巨噬细胞存活率显著升高(p=0.001<0.01),LPS组、蔓菁粗多糖组(200,400μg/mL)巨噬细胞存活率升高(p=0.011,0.021,0.027<0.05);与阴性对照组相比,LPS组与蔓菁粗多糖各剂量组巨噬细胞吞噬指数显著升高(p<0.01),与LPS组相比,蔓菁粗多糖组(50,100,200μg/mL)巨噬细胞吞噬指数显著升高(p<0.01),蔓菁粗多糖组(400μg/mL)巨噬细胞吞噬指数升高(p<0.05);与阴性对照组相比,LPS组及蔓菁粗多糖组(50μg/mL)NO表达量显著升高(p<0.01),与LPS组相比,蔓菁粗多糖组(200μg/mL)NO表达量显著降低(p<0.01),蔓菁粗多糖组(100,400μg/mL)NO表达量降低(p<0.05);与阴性对照组相比,蔓菁粗多糖组(50,100,200μg/mL)脾细胞(T细胞)增殖指数显著升高(p<0.01),conA组脾细胞(T细胞)增殖指数升高(p<0.05),与conA组相比,蔓菁粗多糖组(200μg/mL)脾细胞(T细胞)增殖指数显著升高(p<0.01),蔓菁粗多糖组(50μg/mL)脾细胞(T细胞)增殖指数升高(p<0.05);与阴性对照组相比,蔓菁粗多糖组(100,200,400μg/mL)脾细胞(B细胞)增殖指数显著升高(p<0.01),LPS组及蔓菁粗多糖组(50μg/mL)脾细胞(B细胞)增殖指数升高(p<0.05),与LPS组相比�Exploring the immune activity of RAW264.7 Macrophages affected by extracellular polysaccharides from Manjing through in vitro cell experiments. Took logarithmic growth stage macrophages and adjust the cell concentration to 1.2×10^(5) cells/mL,inoculate on a 96 well plate for 24 h,and set up a negative control group(only complete culture medium added)and a positive control group(LPS,1 μg/mL),the crude polysaccharide group of Manjing(50,100,200,400 μg/mL),after 24 h of cultivation,MTT assay was used to detect cell proliferation activity,phagocytic ability,and NO secretion ability;Mouse cervical dislocation was euthanized,and the spleen was taken in a sterile environment. The cell concentration was adjusted to 1.0×10^(6) mL and inoculated onto a 96 well plate,set up a control group and a crude polysaccharide group(50,100,200,400 μg/mL)and Con A(10 μg/mL)positive control group or LPS(5 μg/mL)positive control group,cultivate for 24,48,72 h. 4 h before the end of cultivation,add MTT(5 mg/mL)10 μL to each well . Centrifuge the supernatant and sufficient shake after adding 100 μL DMSO,measure the absorbance at 570 nm using an enzyme-linked immunosorbent assay(ELISA)reader and calculate the proliferation index. Compared with the negative control group,the group treated with crude polysaccharides from Manjing(50 μg/mL),the survival rate of macrophages significantly increased(p=0.001<0.01),LPS group,Manjing crude polysaccharide group(200,400 μg/mL)the survival rate of macrophages increased(p=0.011,0.021,0.027<0.05);Compared with the negative control group,the phagocytic index of macrophages in the LPS group and the various dose groups of crude polysaccharides from Primulaceae significantly increased(p<0.01),compared with the LPS group,the crude polysaccharide group of Manjing crude polysacchande group(50,100,200 μg/mL),the phagocytic index of macrophages significantly increased(p<0.01),Manjing crude polysaccharide group(400 μg/mL),the phagocytic index of macrophages increased(p<0.05);Compared with the negative

关 键 词：蔓菁粗多糖 增值活性 吞噬能力 免疫调节 

分 类 号：Q255[生物学—细胞生物学]