米根霉发酵松花粉及对3T3-L1细胞分化和脂质代谢的影响  

Fermentation of pine pollen by Rhizopus oryzae and its effect on adipocyte differentiation and lipid metabolism of 3T3-L1 cells

作　　者：王璇 刘其耸 高福凯 何志勇[1] 秦昉[1] 王召君 陈秋铭 陈洁[1,3] 曾茂茂 WANG Xuan;LIU Qisong;GAO Fukai;HE Zhiyong;QIN Fang;WANG Zhaojun;CHEN Qiuming;CHEN Jie;ZENG Maomao(State Key Lab of Food Science and Technology,Jiangnan University,Wuxi 214122,China;Yantai New Era Health Industry Co.Ltd.,Yantai 264000,China;National Engineering Research Center for Functional Food,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学,食品科学与技术国家重点实验室,江苏无锡214122 [2]烟台新时代健康产业有限公司,山东烟台264000 [3]江南大学,国家功能食品工程技术研究中心,江苏无锡214122

出　　处：《食品与发酵工业》2023年第12期1-9,共9页Food and Fermentation Industries

基　　金：国家食品科学与工程一流学科建设项目(JUFSTR20180201)。

摘　　要：为提升松花粉的减脂功效,通过米根霉发酵以增加其活性成分含量。比较了未发酵松花粉(unfermented pine pollen,NFPP)和发酵松花粉(fermented pine pollen,FPP)对3T3-L1细胞分化和脂质代谢的影响并探讨了其作用机制。结果表明,米根霉发酵可以显著增加松花粉中酚类活性成分的含量,5种酚类物质总量从2091.63μg/g提升至3686.62μg/g,其中柚皮素(naringenin,NAR)含量提高了3.27倍(2027.30μg/g)。油红O染色及定量结果表明,FPP能明显减少3T3-L1细胞的脂质积累,同等浓度下优于NFPP。此外,实时荧光定量PCR(quantitative real-time PCR,q-PCR)及蛋白质印迹法(Western blot)结果表明,与NFPP相比,FPP能明显抑制脂肪细胞分化因子CCAAT/增强子结合蛋白α(enhancer binding proteins alpha,C/EBPα)和过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor gamma,PPARγ),上调脂解因子激素敏感性脂肪酶(hormone-sensitive lipase,HSL)和脂肪甘油三酯脂肪酶(adipose triglyceride lipase,ATGL),以及促进脂肪酸氧化因子过氧化物酶体增殖物激活受体α(peroxisome proliferator-activated receptor gamma alpha,PPARα)的表达(P<0.05),可能是由于苷酸活化蛋白激酶(amp-activated protein kinase,AMPK)发生了磷酸化(上调p-AMPKα/AMPKα,p-AMPKβ1/AMPKβ1和p-ACC/ACC的比值)。松花粉及其发酵产物中主要酚类成分NAR可以明显减少蛋白PPARγ、C/EBPα,增加PPARα和ATGL的表达(P<0.05),因此可能在FPP的降脂功效上发挥主要作用。综上所述,FPP在减少3T3-L1细胞脂质积累方面具有更好的效果,可能是由于酚类,特别是NAR的增加。研究结果可为更高降脂活性的松花粉产品的开发提供理论指导。Obesity has become an important global public health problem.Natural products have always been an important direction of obesity prevention research.Pine pollen is a kind of traditional Chinese medicinal and food material,which is rich in phenolics and have activities for lowering lipids and improving lipid metabolism in vivo.However,previous studies have found that most phenolic ingredients in pine pollen exist in the form of binding state,which makes it unable to fully exert its activity of lowering lipids.In order to increase the lipid-reducing effect of pine pollen,fermentation by edible fungus Rhizopus oryzae was employed.Clean and dry pine pollen was used as the sole source of nutrients(substrate)for liquid medium(solid-liquid-ratio 1∶20)and was fermented in flask with 10%(mL∶g)inoculum(containing 106 spores/mL,30℃,180 r/min for 4 days.The effects of unfermented and fermented pine pollen(NFPP,FPP)on 3T3-L1 differentiation and lipid metabolism,including differentiation factors CCAAT enhancer-binding protein alpha(C/EBPα),peroxisome proliferator-activated receptor gamma(PPARγ),liposynthetic factors FAS,lipolysis factors hormone-sensitive lipase(HSL),adipose triglyceride lipase(ATGL)and fatty acid oxidation factor peroxisome proliferator-activated receptor alpha(PPARα),further the related mechanism was investigated.The results of HPLC showed that compared with NFPP,the contents of protocatechuic acid,p-courmaric acid,taxifolin and naringenin were increased by 176.30%,10.42%,88.43%,and 326.75%,respectively,while catechin was decreased by 32.51%.The total levels of five phenolic ingredients in pine pollen increased significantly,from 2091.63μg/g to 3686.62μg/g,and naringenin(NAR)increased from 475.06μg/g to 2027.30μg/g.Oil red O staining and quantitative results showed that FPP could significantly reduce the lipid accumulation in 3T3-L1 cells,which was better than NFPP under the same concentration and at 400μg/mL,the lipid accumulation of FPP was only 57.59%.In addition,the results of q-PCR and Wes

关 键 词：松花粉 发酵 柚皮素 3T3-L1细胞 脂质积累 腺苷酸活化蛋白激 

分 类 号：TS201.4[轻工技术与工程—食品科学]