本文主要研究内容
作者史瑛（2019）在《具有修复抗生素导致肠道生态失调功能的乳杆菌筛选及其机制研究》一文中研究指出：肠道微生物是人体重要的“消化器官”,它和胃肠道相互影响密切并且具有共同进化的共生关系。抗生素大量应用于多种疾病的治疗,但同时也可能会导致肠道微生物群生态失调如关键菌群的缺失、致病菌增多、多样性降低和代谢机能改变等,进而引发肠道疾病的产生,如抗生素相关性腹泻、肠易激综合征、炎症性肠病和伪膜性肠炎等,甚至可能诱发相关代谢、免疫疾病如抗生素相关性肥胖、过敏、哮喘、糖尿病和风湿性关节炎等。已有研究表明,从发酵食品中分离的乳杆菌对肠道健康具有有益作用,并且对肠道的代谢和免疫具有调节功能。本研究使用常见抗生素的包括氨苄青霉素、头孢克肟和克林霉素以建立肠道菌群紊乱模型,并探究不同性质的乳杆菌对肠道菌群、代谢、肠道屏障和免疫的调节功能且深入探究其作用机理,为益生菌的功能性开发提供相应的科学依据。主要研究结果如下:首先通过给小鼠灌胃抗生素(氨苄青霉素、低剂量头孢克肟、高剂量头孢克肟、克林霉素)建立肠道菌群紊乱模型,基于小鼠肠道菌群16S rRNA基因测序表明4种抗生素暴露均导致小鼠微生物群落多样性以及菌群的组成、结构和相对丰度的改变,从而引发小鼠肠道生态和功能失调。利用热图分析和LEfSe算法分析得到氨苄青霉素导致的肠道菌群结构失调中显著改变的关键菌群特征,即相对丰度增加的克雷伯氏菌属(Klebsiella)、厌氧支原体属(Anaeroplasma)、肠球菌属(Enterococcus)和肠杆菌属(Enterobacter),以及相对丰度显著减少的毛螺菌科(Lachnospiraceae)和阿克曼氏菌属(Akkermansia)。采用非量测的多维变量统计(NMDS)得到宿主生理和肠道菌群之间的关联,可知氨苄青霉素导致的菌群失调与血清中内毒素和二胺氧化酶含量增加以及紧密连接蛋白ZO-1的表达下降相关,建立了模拟临床剂量抗生素导致的肠道生态失调的小鼠模型。其次,针对20株分离于发酵食品的乳杆菌进行筛选,得到包括鼠李糖乳杆菌、植物乳杆菌、干酪乳杆菌和瑞士乳杆菌各5株。通过测定它们的生物学性质包括代时、胃肠液耐受能力、糖利用情况、耐药性、细胞粘附性和定植性等,发现乳杆菌的各个性质存在种间和株间差异。不同乳杆菌在模拟胃肠道环境中的耐受能力差异很大,其中,鼠李糖乳杆菌LGG、植物乳杆菌CCFM605和CGMCC12436以及干酪乳杆菌CCFM5和CGMCC12435在模拟胃液和肠液中存活率都相对较高。小鼠体内的定植能力测定中,鼠李糖乳杆菌CCFM492、植物乳杆菌中CCFM4、CCFM595和CGMCC12436以及干酪乳杆菌CCFM30的定植能力良好。综合各体外性质和体内定植进行主成分分析,并选取最接近主成分1和主成分2的菌株,形成两组具有不同优良生物特性的乳杆菌混菌组合,进行后续实验分析。其中乳杆菌混菌A组的菌株细胞粘附率和低聚糖利用性质优良,而乳杆菌混菌B组中的菌株具有相对较快的生长速率以及良好的定植性和模拟肠液耐受性。评价筛选的两组乳杆菌混菌对不同抗生素包括氨苄青霉素、头孢克肟和克林霉素导致肠道菌群紊乱的修复效果,发现乳杆菌混菌A组(包括鼠李糖乳杆菌LGG、植物乳杆菌CCFM4、干酪乳杆菌CCFM5和瑞士乳杆菌CCFM673)和B组(包括鼠李糖乳杆菌CCFM492、植物乳杆菌CGMCC12436、干酪乳杆菌CGMCC12435和瑞士乳杆菌CCFM6)均对氨苄青霉素和高剂量头孢克肟导致的肠道菌群门水平上结构和组成紊乱有恢复作用,但两组乳杆菌混菌对克林霉素导致的紊乱恢复效果不明显。两组乳杆菌混菌对氨苄青霉素和头孢克肟导致的盲肠内容物中短链脂肪酸降低均有显著恢复能力,可提高其含量至正常水平,但是两组乳杆菌混菌对克林霉素导致的短链脂肪酸水平降低的恢复效果较差。此外,乳杆菌混菌B组可以缓解高剂量头孢克肟引发的小鼠结肠和回肠的组织损伤和炎症至对照组水平,且可有效修复高剂量头孢克肟导致的包括Mucispirillum、肠球菌属(Enterococcus)、Alistipes和Turicibacter在内的特定菌群相对丰度的变化。进一步分析乳杆菌单菌株对氨苄青霉素诱导的肠道生态失调的菌群紊乱、代谢物改变和免疫炎症的缓解作用,发现所测8株乳杆菌(来自于乳杆菌混菌A组和B组)对肠道通透性和血清内毒素水平的恢复作用较为普遍。而其中3株菌:干酪乳杆菌CGMCC12435(LacC)、植物乳杆菌CGMCC12436(LacP)和鼠李糖乳杆菌LGG(LacG)可显著增加结肠或回肠中部分紧密连接蛋白如ZO-1、闭合蛋白Occludin和Claudin-1的相对表达量。进一步通过LEfSe分析表明,LacC、和LacP修复组的差异菌群数量比自然恢复组少,说明3个乳杆菌单菌组均对肠道菌群失调有显著修复作用。但在对粪便代谢物的修复效果上3株菌具有差异,仅LacC使粪便中乙酸、丙酸和丁酸的水平显著恢复到初始水平,并且更为有效的恢复了代谢物的整体结构组成。LacC治疗组中肠道菌群和代谢物的相关性分析表明升高的乙酸盐水平和肠道中柠檬酸杆菌属(Citrobacter)、双歧杆菌属(Bifidobacterium)、真杆菌属(Eubacterium)、S24-7、理研菌科(Rikenellaceae)和梭菌科(Clostridiaceae)的相对丰度增加有正相关关系。此外,LacC菌株可有效调节氨苄青霉素导致的结肠组织中sIgA、RegⅢγ和IFN-γ的升高,LacG可降低造模组中sIgA和RegⅢγ水平的升高,而LacP对所测免疫因子无明显调节效果。最后,深入解析乳杆菌在代谢通路、肠道屏障和免疫调节等方面缓解抗生素导致生态失调的机制,结果表明LacC和LacP菌株可显著缓解氨苄青霉素使用后结肠内NF-κB水平的升高,并降低NF-κB p65的免疫荧光表达量,因此这2株菌可通过NF-κB信号通路调节结肠内氨苄青霉素导致的免疫炎症。在Caco-2细胞中通过Transwell构建肠道屏障体系,并使用LPS刺激以探究抗生素作用机制,结果表明乳杆菌LacC和LacP对LPS刺激导致的TEER值下降即肠道屏障破坏的保护效果不明显,但能够调节丁酸受体GPR109A的基因表达上调以激活Toll样受体4信号。根据全基因组分析推测了其中一株植物乳杆菌CGMCC12436的潜在功能性基因,发现其中参与编码荚膜多糖合成的cpsD基因簇,这可能与它良好的肠道适应性和功能活性有关。
Abstract
chang dao wei sheng wu shi ren ti chong yao de “xiao hua qi guan ”,ta he wei chang dao xiang hu ying xiang mi qie bing ju ju you gong tong jin hua de gong sheng guan ji 。kang sheng su da liang ying yong yu duo chong ji bing de zhi liao ,dan tong shi ye ke neng hui dao zhi chang dao wei sheng wu qun sheng tai shi diao ru guan jian jun qun de que shi 、zhi bing jun zeng duo 、duo yang xing jiang di he dai xie ji neng gai bian deng ,jin er yin fa chang dao ji bing de chan sheng ,ru kang sheng su xiang guan xing fu xie 、chang yi ji zeng ge zheng 、yan zheng xing chang bing he wei mo xing chang yan deng ,shen zhi ke neng you fa xiang guan dai xie 、mian yi ji bing ru kang sheng su xiang guan xing fei pan 、guo min 、xiao chuan 、tang niao bing he feng shi xing guan jie yan deng 。yi you yan jiu biao ming ,cong fa jiao shi pin zhong fen li de ru gan jun dui chang dao jian kang ju you you yi zuo yong ,bing ju dui chang dao de dai xie he mian yi ju you diao jie gong neng 。ben yan jiu shi yong chang jian kang sheng su de bao gua an bian qing mei su 、tou bao ke wo he ke lin mei su yi jian li chang dao jun qun wen luan mo xing ,bing tan jiu bu tong xing zhi de ru gan jun dui chang dao jun qun 、dai xie 、chang dao bing zhang he mian yi de diao jie gong neng ju shen ru tan jiu ji zuo yong ji li ,wei yi sheng jun de gong neng xing kai fa di gong xiang ying de ke xue yi ju 。zhu yao yan jiu jie guo ru xia :shou xian tong guo gei xiao shu guan wei kang sheng su (an bian qing mei su 、di ji liang tou bao ke wo 、gao ji liang tou bao ke wo 、ke lin mei su )jian li chang dao jun qun wen luan mo xing ,ji yu xiao shu chang dao jun qun 16S rRNAji yin ce xu biao ming 4chong kang sheng su bao lou jun dao zhi xiao shu wei sheng wu qun la duo yang xing yi ji jun qun de zu cheng 、jie gou he xiang dui feng du de gai bian ,cong er yin fa xiao shu chang dao sheng tai he gong neng shi diao 。li yong re tu fen xi he LEfSesuan fa fen xi de dao an bian qing mei su dao zhi de chang dao jun qun jie gou shi diao zhong xian zhe gai bian de guan jian jun qun te zheng ,ji xiang dui feng du zeng jia de ke lei bai shi jun shu (Klebsiella)、ya yang zhi yuan ti shu (Anaeroplasma)、chang qiu jun shu (Enterococcus)he chang gan jun shu (Enterobacter),yi ji xiang dui feng du xian zhe jian shao de mao luo jun ke (Lachnospiraceae)he a ke man shi jun shu (Akkermansia)。cai yong fei liang ce de duo wei bian liang tong ji (NMDS)de dao su zhu sheng li he chang dao jun qun zhi jian de guan lian ,ke zhi an bian qing mei su dao zhi de jun qun shi diao yu xie qing zhong nei du su he er an yang hua mei han liang zeng jia yi ji jin mi lian jie dan bai ZO-1de biao da xia jiang xiang guan ,jian li le mo ni lin chuang ji liang kang sheng su dao zhi de chang dao sheng tai shi diao de xiao shu mo xing 。ji ci ,zhen dui 20zhu fen li yu fa jiao shi pin de ru gan jun jin hang shai shua ,de dao bao gua shu li tang ru gan jun 、zhi wu ru gan jun 、gan lao ru gan jun he rui shi ru gan jun ge 5zhu 。tong guo ce ding ta men de sheng wu xue xing zhi bao gua dai shi 、wei chang ye nai shou neng li 、tang li yong qing kuang 、nai yao xing 、xi bao nian fu xing he ding zhi xing deng ,fa xian ru gan jun de ge ge xing zhi cun zai chong jian he zhu jian cha yi 。bu tong ru gan jun zai mo ni wei chang dao huan jing zhong de nai shou neng li cha yi hen da ,ji zhong ,shu li tang ru gan jun LGG、zhi wu ru gan jun CCFM605he CGMCC12436yi ji gan lao ru gan jun CCFM5he CGMCC12435zai mo ni wei ye he chang ye zhong cun huo lv dou xiang dui jiao gao 。xiao shu ti nei de ding zhi neng li ce ding zhong ,shu li tang ru gan jun CCFM492、zhi wu ru gan jun zhong CCFM4、CCFM595he CGMCC12436yi ji gan lao ru gan jun CCFM30de ding zhi neng li liang hao 。zeng ge ge ti wai xing zhi he ti nei ding zhi jin hang zhu cheng fen fen xi ,bing shua qu zui jie jin zhu cheng fen 1he zhu cheng fen 2de jun zhu ,xing cheng liang zu ju you bu tong you liang sheng wu te xing de ru gan jun hun jun zu ge ,jin hang hou xu shi yan fen xi 。ji zhong ru gan jun hun jun Azu de jun zhu xi bao nian fu lv he di ju tang li yong xing zhi you liang ,er ru gan jun hun jun Bzu zhong de jun zhu ju you xiang dui jiao kuai de sheng chang su lv yi ji liang hao de ding zhi xing he mo ni chang ye nai shou xing 。ping jia shai shua de liang zu ru gan jun hun jun dui bu tong kang sheng su bao gua an bian qing mei su 、tou bao ke wo he ke lin mei su dao zhi chang dao jun qun wen luan de xiu fu xiao guo ,fa xian ru gan jun hun jun Azu (bao gua shu li tang ru gan jun LGG、zhi wu ru gan jun CCFM4、gan lao ru gan jun CCFM5he rui shi ru gan jun CCFM673)he Bzu (bao gua shu li tang ru gan jun CCFM492、zhi wu ru gan jun CGMCC12436、gan lao ru gan jun CGMCC12435he rui shi ru gan jun CCFM6)jun dui an bian qing mei su he gao ji liang tou bao ke wo dao zhi de chang dao jun qun men shui ping shang jie gou he zu cheng wen luan you hui fu zuo yong ,dan liang zu ru gan jun hun jun dui ke lin mei su dao zhi de wen luan hui fu xiao guo bu ming xian 。liang zu ru gan jun hun jun dui an bian qing mei su he tou bao ke wo dao zhi de mang chang nei rong wu zhong duan lian zhi fang suan jiang di jun you xian zhe hui fu neng li ,ke di gao ji han liang zhi zheng chang shui ping ,dan shi liang zu ru gan jun hun jun dui ke lin mei su dao zhi de duan lian zhi fang suan shui ping jiang di de hui fu xiao guo jiao cha 。ci wai ,ru gan jun hun jun Bzu ke yi huan jie gao ji liang tou bao ke wo yin fa de xiao shu jie chang he hui chang de zu zhi sun shang he yan zheng zhi dui zhao zu shui ping ,ju ke you xiao xiu fu gao ji liang tou bao ke wo dao zhi de bao gua Mucispirillum、chang qiu jun shu (Enterococcus)、Alistipeshe Turicibacterzai nei de te ding jun qun xiang dui feng du de bian hua 。jin yi bu fen xi ru gan jun chan jun zhu dui an bian qing mei su you dao de chang dao sheng tai shi diao de jun qun wen luan 、dai xie wu gai bian he mian yi yan zheng de huan jie zuo yong ,fa xian suo ce 8zhu ru gan jun (lai zi yu ru gan jun hun jun Azu he Bzu )dui chang dao tong tou xing he xie qing nei du su shui ping de hui fu zuo yong jiao wei pu bian 。er ji zhong 3zhu jun :gan lao ru gan jun CGMCC12435(LacC)、zhi wu ru gan jun CGMCC12436(LacP)he shu li tang ru gan jun LGG(LacG)ke xian zhe zeng jia jie chang huo hui chang zhong bu fen jin mi lian jie dan bai ru ZO-1、bi ge dan bai Occludinhe Claudin-1de xiang dui biao da liang 。jin yi bu tong guo LEfSefen xi biao ming ,LacC、he LacPxiu fu zu de cha yi jun qun shu liang bi zi ran hui fu zu shao ,shui ming 3ge ru gan jun chan jun zu jun dui chang dao jun qun shi diao you xian zhe xiu fu zuo yong 。dan zai dui fen bian dai xie wu de xiu fu xiao guo shang 3zhu jun ju you cha yi ,jin LacCshi fen bian zhong yi suan 、bing suan he ding suan de shui ping xian zhe hui fu dao chu shi shui ping ,bing ju geng wei you xiao de hui fu le dai xie wu de zheng ti jie gou zu cheng 。LacCzhi liao zu zhong chang dao jun qun he dai xie wu de xiang guan xing fen xi biao ming sheng gao de yi suan yan shui ping he chang dao zhong ning meng suan gan jun shu (Citrobacter)、shuang qi gan jun shu (Bifidobacterium)、zhen gan jun shu (Eubacterium)、S24-7、li yan jun ke (Rikenellaceae)he suo jun ke (Clostridiaceae)de xiang dui feng du zeng jia you zheng xiang guan guan ji 。ci wai ,LacCjun zhu ke you xiao diao jie an bian qing mei su dao zhi de jie chang zu zhi zhong sIgA、RegⅢγhe IFN-γde sheng gao ,LacGke jiang di zao mo zu zhong sIgAhe RegⅢγshui ping de sheng gao ,er LacPdui suo ce mian yi yin zi mo ming xian diao jie xiao guo 。zui hou ,shen ru jie xi ru gan jun zai dai xie tong lu 、chang dao bing zhang he mian yi diao jie deng fang mian huan jie kang sheng su dao zhi sheng tai shi diao de ji zhi ,jie guo biao ming LacChe LacPjun zhu ke xian zhe huan jie an bian qing mei su shi yong hou jie chang nei NF-κBshui ping de sheng gao ,bing jiang di NF-κB p65de mian yi ying guang biao da liang ,yin ci zhe 2zhu jun ke tong guo NF-κBxin hao tong lu diao jie jie chang nei an bian qing mei su dao zhi de mian yi yan zheng 。zai Caco-2xi bao zhong tong guo Transwellgou jian chang dao bing zhang ti ji ,bing shi yong LPSci ji yi tan jiu kang sheng su zuo yong ji zhi ,jie guo biao ming ru gan jun LacChe LacPdui LPSci ji dao zhi de TEERzhi xia jiang ji chang dao bing zhang po huai de bao hu xiao guo bu ming xian ,dan neng gou diao jie ding suan shou ti GPR109Ade ji yin biao da shang diao yi ji huo Tollyang shou ti 4xin hao 。gen ju quan ji yin zu fen xi tui ce le ji zhong yi zhu zhi wu ru gan jun CGMCC12436de qian zai gong neng xing ji yin ,fa xian ji zhong can yu bian ma jia mo duo tang ge cheng de cpsDji yin cu ,zhe ke neng yu ta liang hao de chang dao kuo ying xing he gong neng huo xing you guan 。
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论文作者分别是来自江南大学的史瑛，发表于刊物江南大学2019-03-18论文，是一篇关于乳杆菌论文,抗生素论文,肠道生态失调论文,微生物组论文,短链脂肪酸论文,免疫屏障论文,江南大学2019-03-18论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自江南大学2019-03-18论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：乳杆菌论文; 抗生素论文; 肠道生态失调论文; 微生物组论文; 短链脂肪酸论文; 免疫屏障论文; 江南大学2019-03-18论文;