不同提取方法的灵芝多糖体外抗炎活性比较  

Comparison of Anti-inflammatory Activity of Ganoderma lucidum Polysaccharides in Vitro by Different Extraction Methods

作　　者：杜小琴 夏炎 张万超[1] 何靖柳[2] 张小琴 吴中宝[1] DU Xiaoqin;XIA Yan;ZHANG Wanchao;HE Jingliu;ZHANG Xiaoqin;WU Zhongbao(Bio-resource Research and Utilization Joint Key Laboratory of Sichuan and Chongqing,Chongqing Institute of Medical Planting Material,Chongqing 408435;Ya'an Polytechnic College,Ya'an 625000,Sichuan)

机构地区：[1]重庆市药物种植研究所特色生物资源研究与利用川渝共建重点实验室,重庆408435 [2]雅安职业技术学院,四川雅安625000

出　　处：《中国食品学报》2025年第2期124-133,共10页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家重点研发计划项目(2021YFD1601005)。

摘　　要：为研究不同提取方法的灵芝多糖(GLP)对脂多糖(LPS)诱导的RAW264.7细胞体外抗炎活性,以回流提取、复合酶法提取、植物乳杆菌发酵+回流提取、植物乳杆菌发酵+复合酶法提取4种方法提取GLP。以LPS诱导建立RAW264.7细胞炎症模型,加入GLP处理24 h,采用CCK-8法检测GLP对RAW264.7细胞生存率的影响,实时荧光定量PCR(qRT-PCR)法检测炎症细胞因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、诱导型一氧化氮合酶(iNOS)的mRNA表达水平,免疫印迹检测JNK/P38 MAPK信号通路蛋白的表达。结果表明:质量浓度在200μg/mL及以下的4种GLP作用于RAW264.7细胞时,细胞存活率为93%以上。与LPS相比,4种方法提取的GLP均可显著(P<0.05)影响RAW264.7细胞炎症因子TNF-α、IL-6、IL-10、iNOS的mRNA表达水平。与回流提取GLP相比,植物乳杆菌发酵+回流提取GLP可显著(P<0.05)抑制TNF-α、IL-6的mRNA表达及促进IL-10的mRNA表达。与另外3种方法提取GLP相比,植物乳杆菌发酵+复合酶法提取GLP可显著(P<0.05)抑制iNOS的mRNA表达水平。与回流提取和复合酶法提取GLP相比,经植物乳杆菌发酵后提取的2种GLP,均能显著(P<0.05)降低P-JNK、P-P38的蛋白表达。GLP具有较好的体外抗炎活性,不同提取方法对GLP体外抗炎活性的影响较大,其中植物乳杆菌发酵后提取的GLP具有更显著的抗炎活性。To investigate the anti-inflammatory activity of Ganoderma lucidum polysaccharides(GLP)in vitro by different extraction methods in RAW264.7 macrophages stimulated by LPS.In this experiment,the extraction of GLP was carried out by hot water,multiple enzyme,fermentation with L.plantarum combined with hot water,fermentation with L.plantarum combined with multiple enzyme.In the process of in vitro experiments,after stimulation of GLP for 24 h,CCK8 assay was used to detect the effect of GLP on the proliferation of RAW264.7 macrophages.The mRNA expression levels of TNF-α,IL-6,IL-10 and iNOS in RAW264.7 macrophages was observed by qRT-PCR.The expression of phosphorylated JNK(P-JNK)and phosphorylated P38 MAPK(P-P38 MAPK)proteins was detected by Western blot.Results indicated that The survival rates of RAW264.7 cells treated with four types of GLP under mass concentration of 200μg/mL were above 93%.Compared with LPS,the four types of GLP exhibited a more significant(P<0.05)influence on the mRNA expression levels of TNF-α,IL-6,IL-10 and iNOS in RAW264.7 macrophages.GLP of fermentation with L.plantarum combined with hot water extraction had a more significant(P<0.05)effect than GLP extracted by hot water alone on the mRNA expression levels of TNF-α,IL-6,IL-10.GLP of fermentation with L.plantarum combined with multiple enzyme extraction had a more significant(P<0.05)effect than GLP extracted by other three extraction methods on the mRNA expression levels of iNOS.GLP of fermentation with L.plantarum had a more significant(P<0.05)effect than GLP extracted solely by hot water and multiple enzyme on the expression levels of JNK/P38 MAPK signal pathway relatived proteins in RAW264.7 cells.This study demonstrated that GLP possesses significant anti-inflammatory activity in vitro,and the extraction methods greatly influenced the anti-inflammatory activity of GLP in vitro.GLP of fermentation with L.plantarum showed a more significant effect than GLP extracted by hot water and multiple enzyme in terms of anti-inflammatory activit

关 键 词：微生物发酵 灵芝多糖 抗炎活性 JNK/P38 MAPK信号通路 

分 类 号：TS219[轻工技术与工程—粮食、油脂及植物蛋白工程]