嗜酸乳杆菌发酵复方中药成分变化对腹泻小鼠的影响
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摘要
为对比研究中药复方经发酵前后其成分变化对小鼠腹泻的影响,本研究采用高效液相色谱检测中药复方发酵前后成分的变化,以大肠杆菌K88所致腹泻小鼠为动物模型,对比研究中药复方发酵液与中药复方水提液对腹泻小鼠腹泻率、腹泻指数、免疫器官指数、肠道菌群及细胞因子IFN-γ、IL-4的影响。结果显示,中药复方发酵液中保留时间为16 min处的主要特征吸收峰消失,主要成分发生了转变,产生了一种新的特征吸收峰。饲喂试验显示发酵组小鼠腹泻指数与腹泻率显著低于中药水提组(p<0.05);中药复方发酵液在降低小鼠盲肠内大肠杆菌数量,提高双歧杆菌及嗜酸乳杆菌数量方面优于中药复方水提液;发酵组小鼠胸腺指数与肝脏指数显著高于水提组(p<0.05)。中药复方发酵液在提高IFN-γ/IL-4比值方面优于中药复方水提液。结果表明,中药复方经过发酵后成分发生了变化,形成了新的物质,对腹泻小鼠产生了有益的影响,应用效果优于中药复方水提液,为其在治疗腹泻方面的应用奠定基础。
To investigate the effect of the composition changes of Traditional Chinese Medicine compound before and after fermentation on diarrhea induced by K88 in mice, we employed HPLC method to detect the changes of components before and after fermentation. The diarrhea mice induced by Escherichia coli K88 were used as animal models to compare the effects of Traditional Chinese Medicine fermentation liquid and Traditional Chinese Medicine water extract on diarrhea rate, diarrhea index,immune organ index, intestinal flora and cytokine IFN-γ/IL-4. The main characteristic absorption peaks of fermentation broth of Traditional Chinese Medicine at 16 min disappeared and the main components changed, resulting in a new characteristic absorption peak. Feeding test showed that the diarrhea index and diarrhea rate of mice in Traditional Chinese Medicine fermentation broth group were significantly lower than those in Traditional Chinese Medicine water extract group(p<0.05). The Traditional Chinese Medicine fermentation broth group was superior to the Traditional Chinese Medicine water extract group in reducing the number of Escherichia coli and increasing the number of Bifidobacterium and lactic acid bacteria in cecum. Thymus index and liver indexwere significantly higher than those of Traditional Chinese Medicine water extract group(p<0.05). The Traditional Chinese Medicine fermentation broth was superior to the Traditional Chinese Medicine water extract group in increasing the ratio of IFN-γ/IL-4. The results showed that the composition of Traditional Chinese Medicine compound was changed after fermentation,which had a beneficial effect on diarrhea mice. The application effect is better than that of Traditional Chinese Medicine compound water extract, which laid a foundation for its application in the treatment of diarrhea.
To investigate the effect of the composition changes of Traditional Chinese Medicine compound before and after fermentation on diarrhea induced by K88 in mice, we employed HPLC method to detect the changes of components before and after fermentation. The diarrhea mice induced by Escherichia coli K88 were used as animal models to compare the effects of Traditional Chinese Medicine fermentation liquid and Traditional Chinese Medicine water extract on diarrhea rate, diarrhea index,immune organ index, intestinal flora and cytokine IFN-γ/IL-4. The main characteristic absorption peaks of fermentation broth of Traditional Chinese Medicine at 16 min disappeared and the main components changed, resulting in a new characteristic absorption peak. Feeding test showed that the diarrhea index and diarrhea rate of mice in Traditional Chinese Medicine fermentation broth group were significantly lower than those in Traditional Chinese Medicine water extract group(p<0.05). The Traditional Chinese Medicine fermentation broth group was superior to the Traditional Chinese Medicine water extract group in reducing the number of Escherichia coli and increasing the number of Bifidobacterium and lactic acid bacteria in cecum. Thymus index and liver indexwere significantly higher than those of Traditional Chinese Medicine water extract group(p<0.05). The Traditional Chinese Medicine fermentation broth was superior to the Traditional Chinese Medicine water extract group in increasing the ratio of IFN-γ/IL-4. The results showed that the composition of Traditional Chinese Medicine compound was changed after fermentation,which had a beneficial effect on diarrhea mice. The application effect is better than that of Traditional Chinese Medicine compound water extract, which laid a foundation for its application in the treatment of diarrhea.
引文
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[17] Chen Zhan-guo, Lu Jiu-ru. Simultaneous and direct determination of oxalic acid, tartaric acid, malic acid, vitamin C, citric acid, and succinic acid in Fructus mume by reversed-phase high-performance liquid chromatography[J]. J Chromatogr Sci,2002, 40(1):35-39.
[18]常连举,张宗和,黄嘉玲,等.没食子酸的制备与应用综述[J].生物质化学工程,2010,44(04):48-52.
[2]刘颖,王彦多,曹广超,等.MTT法比较天龙发酵前后抗肺腺癌A549细胞效果[J].山东化工,2018, 47(09):9-10.
[3] Xing Yon, Cai Le, Yin Tian-peng, et al. Improving the antioxidant activity and enriching salvianolic acids by the fermentationof Salvia miltiorrhizae with Geomyces luteus[J]. J Zhejiang Univ Sci B, 2016, 17(5):391-398.
[4]孙佳彬,覃艺,张红玲,等.发酵对半夏毒性和刺激性的影响[J].中药与临床,2017,8(06):30-33.
[5]梁易晓,刘丽娜,秦顺义,等.羧甲基壳聚糖硒对小鼠生长性能、血液硒含量及抗氧化功能的影响[J].中国饲料,2016,(11):19-21.
[6]刘卫东.不同剂型七味白术散对肠道菌群失调腹泻模型小鼠治疗作用的相关免疫学机理研究[D].长沙:湖南中医药大学,2015.
[7]章丽,刘晓娟,熊平,等.山莓叶醇提物对不同模型小鼠腹泻的影响[J].现代食品科技,2012,28(09):1111-1115.
[8]刘富来,冯翠兰.中药复方制剂防治实验性禽大肠杆菌病的疗效试验[J].中国预防兽医学报,2004,(01):74-78.
[9]马海利,郑明学,李元平,等.中药抗柔嫩艾美尔球虫机制研究[J].中国预防兽医学报,2001, 23(01):34-36.
[10]陈张华,邓俊良,陈仓良.复方中草药“猪康散”对仔猪红细胞免疫功能的影响[J].中国预防兽医学报,2015,37(12):970-972.
[11]韩春杨,刘翠艳,牛钟相.中药制剂发酵前后成分的变化及对肉鸡部分免疫指标和生长的影响[J].畜牧兽医学报,2005,(11):116-120.
[12]仲桢玉.葛根、天花粉发酵过程中化学成分变化研究[D].贵州:贵州医科大学,2018.
[13] Han Chun-chao, Hong Wei. Anti-inflammatory effects of fermented and non-fermented Sophora flavescens:a comparative study[J]. BMC Complement Altern Med, 2011, 11:100.
[14]卢伟.黄芩与鼠李糖乳杆菌发酵产物对仔猪大肠杆菌腹泻治疗效果研究[D].沈阳:吉林农业大学,2016.
[15]李国忠.发酵黄芩对断奶仔猪生长性能和免疫功能的影响[D].哈尔滨:东北农业大学,2012.
[16]刘燕,兰瑞容,兑靖冬,苏英.HPLC法同时测定蒲公英中单咖啡酰酒石酸、绿原酸、咖啡酸、菊苣酸的含量[J].中国药师,2017,20(09):1677-1679.
[17] Chen Zhan-guo, Lu Jiu-ru. Simultaneous and direct determination of oxalic acid, tartaric acid, malic acid, vitamin C, citric acid, and succinic acid in Fructus mume by reversed-phase high-performance liquid chromatography[J]. J Chromatogr Sci,2002, 40(1):35-39.
[18]常连举,张宗和,黄嘉玲,等.没食子酸的制备与应用综述[J].生物质化学工程,2010,44(04):48-52.