Study of Lacticaseibacillus rhamnosus MP108 on Alleviating Bacterial Diarrhea
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摘要: 本文对含鼠李糖乳杆菌MP108产品对肠毒性大肠杆菌引起腹泻的缓解作用进行了评估。将六周龄雄性BALB/c小鼠分为空白组、模型组、益生菌MP108产品组和高剂量益生菌MP108产品组，益生菌干预持续两周。实验结束后，检测小鼠的粪便含水量、血清腹泻相关蛋白及炎症因子、空肠组织病理变化及肠道菌群的变化。结果显示MP108产品组和高剂量MP108产品组均能缓解肠毒性大肠杆菌（Enterotoxic Escherichia coli，ETEC）引起的腹泻症状，减少空肠组织损伤，降低血清炎症的水平，降低了铁蛋白（Serum ferritin，ST）和Toll样受体4（Toll-like receptor 4，TLR4）的表达，同时增加了水通道蛋白3（Aquaporin 3，AQP3）的水平。肠道菌群分析表明，鼠李糖乳杆菌MP108产品有助于恢复肠道的微生态，同时增加产短链脂肪酸的细菌Akkermansia，Ruminococcaceae，Lachnospiraceae的相对丰度。因此，鼠李糖乳杆菌MP108产品可通过调节腹泻相关蛋白、炎症因子的表达以及肠道菌群的变化发挥缓解细菌性腹泻的作用。Abstract: The alleviation effect of Lacticaseibacillus rhamnosus MP108-containing products on diarrhea caused by Enterotoxic Escherichia coli was evaluated. Six-week-old male BALB/c mice were randomly divided into blank group, model group, MP108 probiotic product group, and high-dose MP108 probiotic product group, and probiotic interventions lasted for two weeks. After the experiment, the fecal water content, serum diarrhea-related proteins, inflammatory factors, pathological changes of jejunum tissue, and gut microbial alterations were evaluated. The results showed that both the MP108 product group and the high-dose MP108 product group relieved diarrhea symptoms caused by Enterotoxic Escherichia coli (ETEC) via reducing jejunum damage, decreasing the levels of serum inflammatory factors, suppressing the expressions of serum ferritin (ST) and Toll-like receptor 4 (TLR4), and increasing aquaporin 3 (AQP3) levels. Intestinal flora analysis showed that L. rhamnosus MP108 product restored the intestinal microecology and increased the proportions of short-chain fatty acid-producing bacteria including Akkermansia, Ruminococcaceae, Lachnospiraceae. Collectively, L. rhamnosus MP108 probiotic products relieve diarrhea by regulating the expression of diarrhea-related proteins and inflammatory factors and modulating gut microbiota.
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Key words:
- Lacticaseibacillus rhamnosus /
- Escherichia coli /
- diarrhea /
- inflammatory factors /
- gut microbiota
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图 1 鼠李糖乳杆菌MP108菌株的耐酸耐胆盐能力
Figure 1. Acid and bile salt tolerance of Lacticaseibacillus rhamnosus MP108
注：与对照组比较，采用***（P<0.001）表示差异显著。
图 3 鼠李糖乳杆菌MP108产品对血清中腹泻相关蛋白含量的影响
Figure 3. Effects of L. rhamnosus MP108 product on diarrhea related protein content in serum
注：a免疫球蛋白A（IgA）；b水通道蛋白3（AQP3）；c铁蛋白（ST）；d Toll样受体4（TLR4）。
图 4 鼠李糖乳杆菌MP108产品对血清中细胞因子含量的影响
Figure 4. Effect of L. rhamnosus MP108 product on serum cytokine content
注：a肿瘤坏死因子α（TNF-α）；b-白介素6（IL-6）；c-白介素10（IL-10）；d-干扰素γ（IFN-γ）。
图 5 鼠李糖乳杆菌MP108产品对小鼠肠道菌群的影响
Figure 5. Effects of L. rhamnosus MP108 product on gut microbiota in mice
注：a基于OTU数量衡量菌群的α多样性（Observed）；b基于物种及丰度衡量菌群的α多样性化（Chao1）；c基于均匀度衡量菌群的α多样性化（Shannon）；d基于物种间进化关系衡量菌群的α多样性化（Simpson）；e菌群β多样性；f肠道菌群门水平的变化；g各组差异菌属比较。
表 1 实验分组及处理
Table 1. Experimental groups and treatments
组别 处理 第一周 第二周 第三周 前3 d 后4 d 空白对照组 适应期 生理盐水 链霉素 生理盐水 模型组 生理盐水 链霉素 生理盐水+ETEC MP108产品组 益生菌溶液 链霉素 益生菌溶液+ETEC 高剂量MP108产品组 益生菌溶液 链霉素 益生菌溶液+ETEC 
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[1] LANATA C F, FISCHER-WALKER C L, OLASCOAGA A C, et al. Global causes of diarrheal disease mortality in children <5 years of age: A systematic review[J]. PloS One,2013,8(9):e72788. doi: 10.1371/journal.pone.0072788 [2] WANG H, ZHONG Z, LUO Y, et al. Heat-stable enterotoxins of enterotoxigenic Escherichia coli and their impact on host immunity[J]. Toxins,2019,11(1):24. doi: 10.3390/toxins11010024 [3] VIRK A, MANDREKAR J, BERBARI E F, et al. A randomized, double blind, placebo-controlled trial of an oral synbiotic (AKSB) for prevention of travelers' diarrhea[J]. Journal of Travel Medicine,2013,20(2):88−94. doi: 10.1111/jtm.12008 [4] XU H, ZHAO F, HOU Q, et al. Metagenomic analysis revealed beneficial effects of probiotics in improving the composition and function of the gut microbiota in dogs with diarrhoea[J]. Food & Function,2019,10(5):2618−2629. [5] YANG B, YUE Y, CHEN Y, et al. Lactobacillus plantarum CCFM1143 alleviates chronic diarrhea via inflammation regulation and gut microbiota modulation: A double-blind, randomized, placebo-controlled study[J]. Frontiers in Immunology,2021,12:746585. doi: 10.3389/fimmu.2021.746585 [6] WANG J, JI H, WANG S, et al. Probiotic Lactobacillus plantarum promotesintestinal barrier function by strengthening the epithelium and modulating gut microbiota[J]. Frontiers in Microbiology,2018,9:1953. doi: 10.3389/fmicb.2018.01953 [7] LIU T Y, SONG X L, AN Y P, et al. Lactobacillus rhamnosus GG colonization in early life ameliorates inflammaging of offspring by activating SIRT1/AMPK/PGC-1 alpha pathway[J]. Oxidative Medicine and Cellular Longevity,2021,21(27):3328505. [8] ROSELLI M, FINAMORE A, BRITTI M S, et al. Probiotic bacteria Bifidobacterium animalis MB5 and Lactobacillus rhamnosus GG protect intestinal Caco-2 cells from the inflammation-associated response induced by enterotoxigenic Escherichia coli K88[J]. The British Journal of Nutrition,2006,95(6):1177−1184. doi: 10.1079/BJN20051681 [9] YANG B, HUANG Z, He Z, et al. Protective effect of Bifidobacterium bifidum FSDJN7O5 and Bifidobacterium breve FHNFQ23M3 on diarrhea caused by enterotoxigenic Escherichia coli[J]. Food & Function,2021,12(16):7271−7282. [10] 靳志强, 王延祥, 李平兰, 等. 植物乳杆菌耐酸耐胆盐的体外试验及其降胆固醇作用[J]. 中国食品学报,2009,9(5):24−28. [XING Z Q, WANG Y X, LI P L, et al. Studies on the acid and bile salt resistant ability of Lactobacillus plantarum and its effect of cholesterol reducing in vitro[J]. Journal of Chinese Institute of Food Science and Technology,2009,9(5):24−28. doi: 10.3969/j.issn.1009-7848.2009.05.004 [11] LEE H, KIM J, JEUNG H, et al. Effects of Ficuscarica paste on loperamide-induced constipation in rats[J]. Food and Chemical Toxicology,2012,50(3-4):895−902. doi: 10.1016/j.fct.2011.12.001 [12] BOLYEN E, RIDOUT J R, DILLON M R, et al. Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2[J]. Nature Biotechnology,2019,37(8):852−857. doi: 10.1038/s41587-019-0209-9 [13] NOVAL R M, BURTON O T, WISE P, et al. A microbiota signature associated with experimental food allergy promotes allergic sensitization and anaphylaxis[J]. Journal of Allergy and Clinical Immunology,2013,131(1):201−212. doi: 10.1016/j.jaci.2012.10.026 [14] SEGATA N, IZARD J, WALDRON L, et al. Metagenomic biomarker discovery and explanation[J]. Genome Biology,2011,12(6):R60. doi: 10.1186/gb-2011-12-6-r60 [15] 王春荣, 刘学飞, 程永刚. 肠功能恢复汤对感染性多器官功能障碍综合征大鼠肠黏膜损伤的保护作用[J]. 郑州大学学报(医学版),2008(6):1107−1111. [[WANG C R, LIU X F, CHENG Y G, et al. Protection of intestine function recovery decoction on intestinal mucosa injury for multiple organ dysfunction syndrom rats caused by infection[J]. Joumal of Zhengzhou University (Medircal Sciences),2008(6):1107−1111. doi: 10.13705/j.issn.1671-6825.2008.06.015 [16] 赵凯科, 韩军艳, 郑立, 等. 葎草醇提物对小鼠细菌性腹泻的治疗作用[J]. 中国兽医学报,2015,35(11):1814−1818. [ZHAO K K, HAN J Y, ZHENG L, et al. The rapeutic effect on Humulus scandens alcohol extract on bacterial diarrhea of mice[J]. Chinese Journal of Veterinary Science,2015,35(11):1814−1818. doi: 10.16303/j.cnki.1005-4545.2015.11.17 [17] YANG K M, JIANG Z Y, ZHENG C T, et al. Effect of Lactobacillus plantarum on diarrhea and intestinal barrier function of young piglets challenged with enterotoxigenic Escherichia coli K88[J]. Journal of Animal Science,2014,92(4):1496−1503. doi: 10.2527/jas.2013-6619 [18] XINGTAO Z, DUODUO Z, WUCHENG Q, et al. Exopolysaccharides from Lactobacillus plantarum NCU116 facilitate intestinal homeostasis by modulating intestinal epithelial regeneration and microbiota[J]. Journal of Agricultural and Food Chemistry,2021,69(28):7863−7873. doi: 10.1021/acs.jafc.1c01898 [19] IKARASHI N, KON R, SUGIYAMA K. Aquaporins in the colon as a new therapeutic target in diarrhea and constipation[J]. International Journal of Molecular Sciences,2016,17(7):1172. doi: 10.3390/ijms17071172 [20] PERDIGÓN G, VINTIÑI E, ALVAREZ S, et al. Study of the possible mechanisms involved in the mucosal immune system activation by lactic acid bacteria[J]. Journal of Dairy Science,1999,82(6):1108−1114. doi: 10.3168/jds.S0022-0302(99)75333-6 [21] SOOMRO S, VENKATESWARAN S, VANARSA K, et al. Predicting disease course in ulcerative colitis using stool proteins identified through an aptamer-based screen[J]. Nature Communications,2021,12(1):3989. doi: 10.1038/s41467-021-24235-0 [22] ALVESTEGUI A, OLIVARES-MORALES M, MUÑOZ E, et al. TLR4 participates in the inflammatory response induced by the AAF/II fimbriae from Enteroaggregative Escherichia coli on intestinal epithelial cells[J]. Front Cell Infect Microbiol,2019,9:143. doi: 10.3389/fcimb.2019.00143 [23] 岳月, 杨波, 赵建新, 等. 植物乳杆菌通过肠道菌群调节缓解产肠毒素大肠杆菌导致的腹泻[C]. 第十五届益生菌与健康国际研讨会, 2020: 2.[YUE Y, YANG B, ZHAO J X, et al. Lactobacillus plantarum alleviates diarrhea caused by Enterotoxogenic Escherichia coli by regulating gut microbiota[C]. The 15th International Symposium on Probiotics and Health, 2020: 2. [24] 张睦清, 彪雅宁, 李易水, 等. 车前子对腹泻大鼠小肠组织AQP3基因及蛋白表达的影响[J]. 新中医,2021,53(17):1−5. [[ZHANG M Q, BIAO Y N, LI Y S, et al. Effect of semen plantaginis on expression of AQP3 gene and protein in small intestine of diarrhea rats[J]. New Chinese Medicine,2021,53(17):1−5. doi: 10.13457/j.cnki.jncm.2021.17.001 [25] 丁晓洁, 孙喜灵, 于晓飞, 等. 乌梅丸对腹泻型肠易激综合征模型大鼠肠道菌群和炎症因子的影响[J]. 辽宁中医杂志,2019,46(6):1296−1299. [DING X J, SUN X L, YU X F, et al. Effect of Wumei pill on intestinal flora and inflammatory factors in rats with diarrhea-type irritable bowel syndrome[J]. Liaoning Traditional Chinese Medicine Journal,2019,46(6):1296−1299. doi: 10.13192/j.issn.1000-1719.2019.06.056 [26] CASTRO-MEJIA J L, O'FERRALL S, KRYCHA L, et al. Restitution of gut microbiota in Ugandan children administered with probiotics (Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12) during treatment for severe acute malnutrition[J]. Gut Microbes,2020,11(4):13. [27] BIN P, TANG Z, LIU S, et al. Intestinal microbiota mediates enterotoxigenic Escherichia coli-induced diarrhea in piglets[J]. BMC Veterinary Research,2018,14(1):385. doi: 10.1186/s12917-018-1704-9 [28] LIU S M, LI E Y, SUN Z Y, et al. Altered gut microbiota and short chain fatty acids in Chinese children with autism spectrum disorder[J]. Scientific Reports,2019,9(1):287. doi: 10.1038/s41598-018-36430-z [29] LIU C, LIANG X, WEI X, et al. Gegen Qinlian decoction treats diarrhea in piglets by modulating gut microbiota and short-chain fatty acids[J]. Frontiers in Microbiology,2019,10:825. doi: 10.3389/fmicb.2019.00825 [30] YU Y, LU J, SUN L, et al. Akkermansia muciniphila: A potential novel mechanism of nuciferine to improve hyperlipidemia[J]. Biomedicine Pharmacotherapy,2021,133:111014. doi: 10.1016/j.biopha.2020.111014 [31] LÓPEZ-COLOM P, CASTILLEJOS L, RODRIGUEZ-SORRENTO A, et al. Impact of in-feed sodium butyrate or sodium heptanoate protected with medium-chain fatty acids on gut health in weaned piglets challenged with Escherichia coli F4+[J]. Archives of Animal Nutrition,2020,74(4):271−295. doi: 10.1080/1745039X.2020.1726719 [32] WANG Q, HE Y, LI X, et al. Lactobacillus reuteri CCFM8631 alleviates hypercholesterolaemia caused by the Paigen atherogenic diet by regulating the gut microbiota[J]. Nutrients,2022,14(6):1272. doi: 10.3390/nu14061272 [33] MOORTHY M, WIE CC, MARIÑO E, et al. The prebiotic potential of geraniin and geraniin-enriched extract against high-fat-diet-induced metabolic syndrome in spraguedawleyrats[J]. Antioxidants (Basel),2022,11(4):632. doi: 10.3390/antiox11040632 -
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