长双歧杆菌评价淫羊藿总黄酮益生元生物活性研究  

Evaluation of probiotic bioactivity of epimedium total flavonoids based on Bifidobacterium longum

作　　者：郭强 王嘉歆 马骁驰 田象阁 黄慧莲 GUO Qiang;WANG Jiaxin;MA Xiaochi;TIAN Xiangge;HUANG Huilian(Laboratory of Modern Preparation of TCM,Ministry of Education,Jiangxi University of Traditional Chinese Medicine,Nanchang,JiangXi 330004,China;不详)

机构地区：[1]江西中医药大学现代中药制剂教育部重点实验室,江西南昌330004 [2]大连医科大学附属第二医院药学研究中心

出　　处：《中国微生态学杂志》2024年第6期621-628,共8页Chinese Journal of Microecology

基　　金：江西中医药大学校级科技创新团队发展计划(CXTD22002);中药改良创新江西省重点实验室支持(2024SSY07131)。

摘　　要：目的 研究淫羊藿总黄酮(total flavonoids of epimedium,TFE)对长双歧杆菌生长代谢的调控影响与长双歧杆菌发酵液的抗炎活性。方法 葡萄糖缺乏的MRS培养基中分别加入浓度为0.5%、1.0%、2.0%的淫羊藿总黄酮作为培养基,设置2.0%葡萄糖和2.0%低聚果糖培养基作为对照,测定发酵后长双歧杆菌的活菌数(CFU)、吸光度值等指标,以明确淫羊藿总黄酮对长双歧杆菌生长的影响,并使用1,1-二苯基-2-三硝基苯肼(DPPH)测定发酵液的体外抗氧化活性。以LPS诱导的RAW264.7为炎症模型,评价培养前后发酵液对造模前后相关炎症因子水平以确定抗炎作用。结果TFE能够显著促进长双歧杆菌的增殖,5 mg/mL、0.5 mg/mL、0.05 mg/mL TFE的促生长率分别为59%、23%、20%,24 h生长曲线也表明TFE的促增殖作用。胃肠道耐受性能研究发现,在长双歧杆菌生长环境中添加2%TFE能够提高菌株在0.3%胆盐(t=6.008,P<0.01)、人工胃液(t=4.371,P<0.05)和pH值为3的酸性环境(t=9.645,P<0.001)中的稳定性,并提高了菌株在含有氯四环素、克林霉素、卡那霉素、甲硝唑、万古霉素的培养基中的生长浓度和体外抗氧化能力(t=15.141,P<0.001)。此外,添加TFE的长双歧杆菌发酵液可降低LPS诱导下的巨噬细胞表达促炎因子TNF-α(t=4.655,P<0.01)、IL-1β(t=7.775,P<0.01)和IL-6(t=7.775,P<0.01)的水平,进而改善LPS诱导的炎症反应。结论 淫羊藿总黄酮可促进长双歧杆菌的生长与菌株稳定性,并增强长双歧杆菌的抗炎活性,有成为新型中药来源益生元的潜力。Objective To observe the regulatory effects of total flavonoids of epimedium(TFE)on the growth and metabolism of Bifidobacterium longum and the anti-inflammatory activity of Bifidobacterium longum fermentation broth.Methods TFE at concentrations of 0.5%,1.0%and 2.0%was added to a glucose-deficient MRS medium as the culture base.With the media containing 2.0%glucose and 2.0%fructooligosaccharides as the controls,the colony forming unit(CFU),OD value and other indicators of Bifidobacterium longum were measured after fermentation to assess the impact of TFE on the growth of Bifidobacterium longum and the in vitro antioxidant activity of the fermentation broth was measured using 1,1-diphenyl-2-picrylhydrazyl(DPPH).RAW264.7 cells induced by LPS were used as an inflammation model to evaluate the anti-inflammatory activity of the fermentation broth.The levels of relevant inflammatory factors before and after fermentation were assessed to determine the anti-inflammatory effect.Results TFE significantly promoted the proliferation of Bifidobacterium longum.The growth promoting rates of different concentrations of TFE(5 mg/mL,0.5 mg/mL and 0.05 mg/mL)compared to the control group were 59%,23%,and 20%,respectively.The 24-hour growth curve also indicated the proliferative effect of TFE.Research on gastrointestinal tolerance found that adding 2%TFE to the growth environment of Bifidobacterium longum can improve the stability of the strain in 0.3%bile salt(t=6.008,P<0.01),artificial gastric juice(t=4.371,P<0.05),and pH3 acidic environment(t=9.645,P<0.001),and increased the growth concentration and in vitro antioxidant capacity of the strains in culture media containing chlortetracycline,clindamycin,kanamycin,metronidazole,and vancomycin(t=15.141,P<0.001).In addition,adding TFE to the fermentation broth of Bifidobacterium longum reduced the levels of pro-inflammatory factors TNF-α(t=4.655,P<0.01),IL-1β(t=7.775,P<0.01),and IL-6(t=7.775,P<0.01),thus improved LPS induced inflammatory response.Conclusion TFE can promote the growth

关 键 词：淫羊藿总黄酮 长双歧杆菌 益生元 抗炎 

分 类 号：R932[医药卫生—生药学]