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Changes in Intracellular and Extracellular Metabolites of Mixed Lactobacillus Strains Enhance Inhibition of Pathogenic Bacterial Growth and Lipopolysaccharide-Induced Alleviation of RAW264.7 Cellular Inflammation

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Abstract

It is important to explore whether there are antagonistic and synergistic effects between different strains of Lactobacillus when developing mixed Lactobacillus strain products. In this study, we investigated the antagonistic and symbiotic effects of co-cultured Lactobacillus strains, as well as their amelioratory effects on lipopolysaccharide (LPS)-induced inflammation and oxidative stress in RAW264.7 cells. The Lactobacillus strains tested in this paper showed no antagonism. Co-culture of Lactiplantibacillus plantarum Y44 and L. plantarum AKS-WS9 was found to show inhibiting effects on the growth of Escherichia coli and Staphylococcus aureus. Additionally, the co-cultured Lactiplantibacillus plantarum Y44 and L. plantarum AKS-WS9 relieved inflammation in RAW264.7 cells induced by LPS by inhibiting the activation of NF-κB and P38 signaling pathways and down-regulating the expression of pro-inflammatory cytokines NO, ROS, iNOs and TNF-α. And the co-cultured Lactobacillus strains activated the Nrf2 signaling pathway in the LPS-induced RAW264.7 cells to promote the expression of antioxidant enzymes in response to oxidative stress. There was a difference in intracellular and extracellular metabolites between single or co-cultured Lactobacillus strains, and the co-cultured Lactobacillus strains significantly increased extracellular metabolites 4-chlorobenzaldehyde, psoromic acid, and 2-dodecylbenzenesulfonic acid and intracellular metabolites 9(S)-HODE, pyocyanin, and LysoPA. We inferred that the better antibacterial and anti-inflammatory ability of the co-cultured Lactobacillus strains were related to the changes in the metabolites of the co-cultured Lactobacillus strains. The co-cultured L. plantarum Y44 and L. plantarum AKS-WS9 strains exhibited better anti-inflammatory abilities and had the potential to alleviate the symptoms of inflammatory diseases as mixed probiotics.

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References

  1. Khani V, Hosseini HM, Taheri M, Nourani MR, Abbas A (2012) Probiotics as an alternative strategy for prevention and treatment of human diseases: a review. Inflamm Allergy - Drug Targets 11(2):79–89. https://doi.org/10.2174/187152812800392832

    Article  CAS  PubMed  Google Scholar 

  2. Barakat OS, El G (2011) Identification and probiotic characteristics of Lactobacillus strains isolated from traditional domiati cheese. Int J Microbiol 3(1):59–66. https://doi.org/10.9735/0975-5276.3.1.59-66

  3. Ouwehand AC, Salminen S, Isolauri E (2002) Probiotics: an overview of beneficial effects. Antonie Van 82(1–4):279–289. https://doi.org/10.1023/A:1020620607611

  4. Zhang ZG, Li Y, Ng CT, Song YQ (2015) Inflammation in Alzheimer’s disease and molecular genetics: recent update. Arch Immune Ther Ex 63(5):333–344. https://doi.org/10.1007/s00005-015-0351-0

    Article  CAS  Google Scholar 

  5. Tuo YF, Song X, Song YL et al (2018) Screening probiotics from Lactobacillus strains according to their abilities to inhibit pathogen adhesion and induction of pro-inflammatory cytokine IL-8. Enl 101(6):4822–4829. https://doi.org/10.3168/jds.2017-13654

    Article  CAS  Google Scholar 

  6. Hampe CS, Roth CL (2017) Probiotic strains and mechanistic insights for the treatment of type 2 diabetes. Endocrine 58(2):207–227. https://doi.org/10.1007/s12020-017-1433-z

    Article  CAS  PubMed  Google Scholar 

  7. Jang SE, Jeong JJ, Kim JK et al (2018) Simultaneous amelioration of colitis and liver injury in mice by Bifidobacterium longum LC67 and Lactobacillus plantarum LC27. Sci Rep 8(1):7500. https://doi.org/10.1038/s41598-018-25775-0

  8. Wang YY, Guo YL, Chen H et al (2018) Potential of Lactobacillus plantarum ZDY2013 and Bifidobacterium bifidum WBIN03 in relieving colitis by gut microbiota, immune and anti-oxidative stress. Can J Microbiol 64(5):327–337. https://doi.org/10.1139/cjm-2017-0716

    Article  CAS  PubMed  Google Scholar 

  9. Huang RH, Tao XY, Wan CX et al (2015) In vitro probiotic characteristics of Lactobacillus plantarum ZDY 2013 and its modulatory effect on gut microbiota of mice. J Dairy Sci 98(9):5850–5861. https://doi.org/10.3168/jds.2014-9153

    Article  CAS  PubMed  Google Scholar 

  10. Wang J, Wu T, Fang L et al (2020) Peptides from walnut (juglans mandshurica maxim.) protect hepatic hepg2 cells from high glucose-induced insulin resistance and oxidative stress. Food Funct 11(9):8112–8121. https://doi.org/10.1039/D0FO01753A

  11. Khandrika L, Kumar B, Koul S et al (2009) Oxidative stress in prostate cancer. Cancer Lett 282(2):125–36. https://doi.org/10.1016/j.canlet.2008.12.011

  12. Soraya S, Mozafar K (2017) Oxidative stress and cancer: the role of Nrf2. CurrCancer Drug Tar 17(6):538–557. https://doi.org/10.2174/1568009617666171002144228

    Article  CAS  Google Scholar 

  13. Li ST, Dai Q, Zhang SX et al (2018) Ulinastatin attenuates LPS-induced inflammation in mouse macrophage RAW264.7 cells by inhibiting the JNK/NF-κB signaling pathway and activating the PI3K/Akt/Nrf2 pathway. Acta Pharmacol Sin 39(8):1294–1304. https://doi.org/10.1038/aps.2017.143

  14. Han S, Gao H, Chen S et al (2019) Procyanidin A1 alleviates inflammatory Response induced by LPS through NF-κB, MAPK, and Nrf 2/HO-1 pathways in RAW264.7 cells. Sci Rep 9(1):15087. https://doi.org/10.1038/s41598-019-51614-x

  15. Li XY, Liu R, Zhao YM et al (2020) The extract from the roots of Rose odorata sweet var. gigantean (Coll. et Hemsl.) Rehd. et Wils attenuates DSS-induced ulcerative colitis by regulating the Nrf2/NF-κB signaling pathways. Rsc Adv 10(16):9450–9461. https://doi.org/10.1039/C9RA10747A

  16. Azad MAK, Sarker M, Wan D (2018) Immunomodulatory effects of probiotics on cytokine profiles. Biomed Res Int 2018:8063647. https://doi.org/10.1155/2018/8063647

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Almutairi B, Turner MS, Fletcher MT, Sultanbawa Y (2020) The impact of commercial prebiotics on the growth, survival and nisin production by Lactococcus lactis 537 in milk. LWT 137:110356. https://doi.org/10.1016/j.lwt.2020.110356

  18. Gao Y, Liu YJ, Ma FL et al (2021) Lactobacillus plantarum Y44 alleviates oxidative stress by regulating gut microbiota and colonic barrier function in Balb/C mice with subcutaneous D-galactose injection. Food Funct 12(1):373–386. https://doi.org/10.1039/D0FO02794D

  19. Liu YJ, Gao Y, Ma FL et al (2020) The ameliorative effect of Lactobacillus plantarum Y44 oral administration on inflammation and lipid metabolism in obese mice fed with a high fat diet. Food Funct 11(6):5024–5039. https://doi.org/10.1007/s10529-020-02895-0

  20. Mu GQ, Li HY, Tuo YF, Gao Y, Zhang YQ (2019) Antioxidative effect of Lactobacillus plantarum Y44 on 2,2’-azobis(2-methylpropionamidine) dihydrochloride (ABAP)-damaged Caco-2 cells. J Dairy Sci 102(8):6863–6875. https://doi.org/10.3168/jds.2019-16447

    Article  CAS  PubMed  Google Scholar 

  21. Wang AR, Li PP, Ma FL, Li XL, Tuo YF, Mu GQ (2023) Mixed Lactiplantibacillus plantarum strains alleviated DSS-induced intestinal inflammation of Balb/c mice via the 5-HT/5-HT7R/NF-κB signaling pathway. J Funct Foods 102(2023)105435. https://doi.org/10.1016/j.jff.2023.105435

  22. Wang SG, Yao JY, Zhou B et al (2018) Bacteriostatic effect of quercetin as an antibiotic alternative in vivo and its antibacterial mechanism in vitro. JFP 81(1):68–78. https://doi.org/10.4315/0362-028X.JFP-17-214

    Article  CAS  PubMed  Google Scholar 

  23. Choi SH, Lee SH, Kim MG et al (2019) Lactobacillus plantarum CAU1055 ameliorates inflammation in lipopolysaccharide-induced RAW264.7 cells and a dextran sulfate sodium-induced colitis animal model. J Dairy Sci 102(8):6718–6725. https://doi.org/10.3168/jds.2018-16197

  24. Jia GC, Liu XF, Che N et al (2020) Human-origin Lactobacillusus salivarius AR809 protects against immunosuppression in S. aureus-induced pharyngitis via Akt-mediated NF-κB and autophagy signaling pathways. Food funct 11(1):270–284. https://doi.org/10.1039/C9FO02476J

  25. Kobayashi EH, Suzuki T, Funayama R et al (2016) Nrf2 suppresses macrophage inflammatory response by blocking proinflammatory cytokine transcription. Nat Commun 7:11624. https://doi.org/10.1038/ncomms11624

  26. Wang AR, Hou KR, Mu GQ et al (2021) Antioxidative effect of soybean milk fermented by Lactobacillus plantarum Y16 on 2, 2 –azobis (2-methylpropionamidine) dihydrochloride (ABAP)-damaged HepG2 cells. FBRC 44:2212–4292. https://doi.org/10.1016/j.fbio.2021.101120

    Article  CAS  Google Scholar 

  27. Peng H, Zhou G, Yang XM et al (2022) Transcriptomic analysis revealed antimicrobial mechanisms of Lactobacillus rhamnosus SCB0119 against Escherichia coli and Staphylococcus aureus. Int J Mol Sci 23(23):15159. https://doi.org/10.3390/ijms232315159.

  28. Kathayat D, Closs G Jr, Helmy YA (2022) In vitro and in vivo evaluation of Lacticaseibacillus rhamnosus GG and Bifidobacterium lactis Bb12 against avian pathogenic Escherichia coli and identification of novel probiotic-derived bioactive peptides. Probiotics Antimicrob Proteins 14(6):1012–1028. https://doi.org/10.1007/s12602-021-09840-1.

  29. Schetter AJ, Heegaard NH, Harris CC (2010) Inflammation and cancer: interweaving microRNA, free radical, cytokine and p53 pathways. Carcinogenesis 31(1):37–49. https://doi.org/10.1093/carcin/bgp272

    Article  CAS  PubMed  Google Scholar 

  30. Aggarwal BB, Vijayalekshmi RV, Sung B (2009) Targeting inflammatory pathways for prevention and therapy of cancer: short-term friend, long-term foe. ClinCancer Res 15(2):425–430. https://doi.org/10.1158/1078-0432.CCR-08-0149

    Article  CAS  Google Scholar 

  31. Lee N, Lee S, Jang SW et al (2021) Lysed and disrupted Bifidobacterium bifidum BGN4 cells promote anti-inflammatory activities in lipopolysaccharide-stimulated RAW 264.7 cells. Saudi Journal Biol Sci 28(9):5115–5118. https://doi.org/10.1016/j.sjbs.2021.05.028

  32. Tanoue T, Nishitani Y, Kanazawa K et al (2008) In vitro model to estimate gut inflammation using co-cultured Caco-2 and RAW264.7 cells. BiochemBioph Res Co 374(3):565–569. https://doi.org/10.1016/j.bbrc.2008.07.063

  33. Kim WJ, Yu HS, Lee NK, Paik HD (2022) Levilactobacillus brevis KU15151 inhibits Staphylococcus aureus lipoteichoic acid-induced inflammation in RAW 264.7 macrophages. Probiotics Antimicrob Proteins 14(4):767–777. https://doi.org/10.1007/s12602-022-09949-x.

  34. Luo G, Kong J, Chi-Yan Cheng B et al (2019) Xiao Qing Long Tang essential oil exhibits inhibitory effects on the release of pro-inflammatory mediators by suppressing NF-κB, AP-1, and IRF3 signaling in the lipopolysaccharide-stimulated RAW264.7 cells. Rsc Adv 9(23):12977–12989. https://doi.org/10.1039/C9RA01448A

  35. Zhou XR, Liu HZ, Zhang J et al (2019) Protective effect of Lactobacillus fermentium CQPC04 on dextran sulfate sodium–induced colitis in mice is associated with modulation of the nuclear factor-κB signaling pathway- sciencedirect. J Dairy Sci 102(11):9570–9585. https://doi.org/10.3168/jds.2019-16840

    Article  CAS  PubMed  Google Scholar 

  36. Shi J, Xie Q, Li H et al (2020) Selected Lactobacillus strains inhibit inflammation in LPS-induced RAW264.7 macrophages by suppressing the TLR4-mediated NF-κB and MAPKs activation. Food Sci Tech-Brazil. https://doi.org/10.21203/rs.3.rs-127575/v1.

  37. Xue EX, Lin JP, Zhang Y et al (2017) Pterostilbene inhibits inflammation and ROS production in chondrocytes by activating Nrf2 pathway. Oncotarget 8(26):41988–42000. https://doi.org/10.18632/oncotarget.16716

  38. Xu H, Wang J, Cai J et al (2019) Protective effect of Lactobacillus rhamnosus GG and its supernatant against myocardial dysfunction in obese mice exposed to intermittent hypoxia is associated with the activation of Nrf2 pathway. Int J Biol Sci 15(11):2471–2483. https://doi.org/10.7150/ijbs.36465

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Toumi R, Soufli I, Rafa H et al (2014) Probiotic bacteria Lactobacillus and Bifidobacterium attenuate inflammation in dextran sulfate sodium-induced experimental colitis in mice. Internet J Immunopathol Pharmacol 27(4):615–627. https://doi.org/10.1177/039463201402700418

    Article  CAS  Google Scholar 

  40. Lim SM, Jeong JJ, Jang SE et al (2016) A mixture of the probiotic strains Bifidobacterium longum CH57 and Lactobacillus brevis CH23 ameliorates colitis in mice by inhibiting macrophage activation and restoring the th17/treg balance. J Funct Foods 27:295–309. https://doi.org/10.1016/j.jff.2016.09.011

  41. Yellamanda B, Vijayalakshmi M, Kavitha A et al (2016) Extraction and bioactive profile of the mixed produced by Rhodococcus sp. VLD-10. 3 Biotech 6(2):261. https://doi.org/10.1007/s13205-016-0576-6

  42. Sweidan A, Chollet-Krugler M, Sauvager A et al (2017) Antibacterial activities of natural lichen compounds against Streptococcus gordonii and Porphyromonas gingivalis. Fitoterapia 121:164–169. https://doi.org/10.1016/j.fitote.2017.07.011

    Article  CAS  PubMed  Google Scholar 

  43. Ali SM, Khan NA, Sagathevan K et al (2019) Biologically active metabolite(s) from haemolymph of red-headed centipede Scolopendra subspinipes possess broad spectrum antibacterial activity. AMB Express 9(1):95. https://doi.org/10.1186/s13568-019-0816-3

  44. Marreiro de Sales-Neto J, Lima ÉA, Cavalcante-Silva LHA et al (2019) Anti-inflammatory potential of pyocyanin in LPS-stimulated murine macrophages. Immunopharmacol Immunotoxicol 41(1):102–108. https://doi.org/10.1080/08923973.2018.1555845

    Article  CAS  PubMed  Google Scholar 

  45. Rolin J, Vego H, Maghazachi AA (2014) Oxidized lipids and lysophosphatidylcholine induce the chemotaxis, up-regulate the expression of CCR9 and CXCR4 and abrogate the release of IL-6 in human monocytes. Toxins (Basel) 6(9):2840–56. https://doi.org/10.3390/toxins6092840

  46. de Vries B, Matthijsen RA, van Bijnen AA et al (2003) Lysophosphatidic acid prevents renal ischemia-reperfusion injury by inhibition of apoptosis and complement activation. Am J Pathol 163(1):47–56. https://doi.org/10.1016/S0002-9440(10)63629-2

  47. Kim SH, Song JH, Kim J, Kang DK (2020) Characterisation of a lysophospholipase from Lactobacillus mucosae. Biotechnol Lett 42(9):1735–1741. https://doi.org/10.1007/s10529-020-02895-0

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This research project was supported by the National Key Research and Development Plan (2021YFD2100700).

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Arong Wang: conceptualization, methodology, formal analysis, writing—original draft, visualization. Chen Guan: methodology, investigation. Tieqi Wang: methodology, investigation. Guangqing Mu: funding acquisition, project administration, supervision. Yanfeng Tuo: funding acquisition, project administration, supervision.

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Correspondence to Yanfeng Tuo.

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Wang, A., Guan, C., Wang, T. et al. Changes in Intracellular and Extracellular Metabolites of Mixed Lactobacillus Strains Enhance Inhibition of Pathogenic Bacterial Growth and Lipopolysaccharide-Induced Alleviation of RAW264.7 Cellular Inflammation. Probiotics & Antimicro. Prot. (2023). https://doi.org/10.1007/s12602-023-10132-z

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