Lactobacillus acidophilus Supplementation Restores Gut Epithelial Integrities and Barrier Functions in Non-specific Diarrhea

Year 2022, , 54 - 62, 15.06.2022

Hossain Ferdaus Mohd Altaf Sumaiya Islam Farzana Khan Parvez Zahran Eman Atif Shaikh Muhammad Uddin Md Nazım Zinnah Kazi Mohammad Ali Khan Imdad Ullah Rahman Md Masudur

https://doi.org/10.5799/jmid.1130109

Abstract

Objectives: The study was aimed to evaluate the roles of Lactobacillus acidophilus supplementation in maintaining intestinal epithelial integrities, tight junction proteins, and adhesion molecules in non-specific diarrhea.
Methods: In this study, we used the pre-weaned BL/6 pups (3 weeks of age, same litters) as a model animal. We supplied the non-sterilized and poor-quality water to experimental pups (n=7) to develop non-specific diarrheic symptoms. Then diarrheic pups were supplemented with Lactobacillus acidophilus (LA) for three consecutive weeks. The control group (n=5) was supplied with sterilized water and no LA. The sampling and analysis were performed on day 0, day 7, day 14, and day 21. The expressions of pro-inflammatory cytokines (IL-6, TNF-a) and tight junction proteins (TJPs) of gut mucosa were determined using qRT-PCR. And the serum cytokines level was screened through sandwich ELISA.
Results: The intestinal cytoskeletal integrity becomes disrupted and characterized by lower ZO-1, Occludin, Claudin-1, Claudin-5, and JAM mRNA expressions upon real-time qRT-PCR. However, Claudin-4 was found to be not affected and illustrated with a higher expression like control pups. Interestingly, supplementing Lactobacillus acidophilus was found to maintain gut integrity and effectively reduce diarrheic symptoms. Like the control pups, the Lactobacillus acidophilus supplemented pups exhibited a higher expression of gut epithelial TJPs and adhesion molecules. Moreover, the diseased pups produced significantly increased IL-6, and TNF-α production in blood serum, compared to control BL/6 pups.
Conclusion: We concluded that L. acidophilus supplementation might orchestrate the equilibrium of gut health and immunity against non-specific diarrhea.

Keywords

mice, gut immunity, tight junctions, and adhesion molecules, Lactobacillus acidophilus supplementatio

References

• 1. Wang J, Linnenbrink M, Kunzel S, et al. Dietary history contributes to enterotype-like clustering and functional metagenomic content in the intestinal microbiome of wild mice. Proc Natl Acad Sci USA 2014, 111(26): 2703-2710.
• 2. Lee SH. Intestinal permeability regulation by tight junction: implication on inflammatory bowel diseases. Intest Res 2015; 13(1): 11-18.
• 3. Turner JR. Intestinal mucosal barrier function in health and disease. Nat Rev Immunol 2009; 9(11): 799-809.
• 4. Gonzalez-Mariscal L, Betanzos A, Nava P, et al. Tight junction proteins. Prog Biophys Mol Biol 2003; 81(1): 1-44.
• 5. Montesi A, Garcia-Albiach R, Pozuelo MJ, et al. Molecular and microbiological analysis of caecal microbiota in rats fed with diets supplemented either with prebiotics or probiotics. Int J Food Microbiol 2005; 98(3): 281-289.
• 6. Johnson AM, Kaushik RS, Hardwidge PR. Disruption of transepithelial resistance by enterotoxigenic Escherichia coli. Vet Microbiol 2010; 141(1-2): 115-119.
• 7. Chung H, Kasper DL. Microbiota-stimulated immune mechanisms to maintain gut homeostasis. Curr Opin Immunol 2010; 22(4): 455-460.
• 8. Rousseaux C, Thuru X, Gelot A, et al. Lactobacillus acidophilus modulates intestinal pain and induces opioid and cannabinoid receptors. Nat Med 2007; 13(1): 35-37.
• 9. Wong JM, de Souza R, Kendall CW, et al. Colonic health: fermentation and short chain fatty acids. J Clin Gastroenterol 2006; 40(3): 235-243.
• 10. Gaschott T, Steinhilber D, Milovic V, et al. Tributyrin, a stable and rapidly absorbed prodrug of butyric acid, enhances antiproliferative effects of dihydroxycholecalciferol in human colon cancer cells. J Nutr 2001; 131(6): 1839-1843.
• 11. Savkovic SD, Villanueva J, Turner JR, et al. Mouse model of enteropathogenic Escherichia coli infection. Infect Immun 2005; 73(2): 1161-1170.
• 12. Mohawk KL, O'Brien AD. Mouse models of Escherichia coli O157:H7 infection and shiga toxin injection. J Biomed Biotechnol 2011; 2011: 258185.
• 13. Yoseph BP, Klingensmith NJ, Liang Z, et al. Mechanisms of Intestinal Barrier Dysfunction in Sepsis. Shock 2016; 46(1):52-59.
• 14. Hossain FMA, Park SO, Kim HJ, et al. CCR5 attenuates neutrophilic airway inflammation exacerbated by infection with rhinovirus. Cell Immunol 2020 351:104066.
• 15. Suzuki T. Regulation of intestinal epithelial permeability by tight junctions. Cell Mol Life Sci 2013; 70(4): 631-659.
• 16. Mennigen R, Nolte K, Rijcken E, et al. Probiotic mixture VSL#3 protects the epithelial barrier by maintaining tight junction protein expression and preventing apoptosis in a murine model of colitis. Am J Physiol Gastrointest Liver Physiol 2009; 296(5): 1140-1149.
• 17. Schultz M, Linde HJ, Lehn N, et al. Immunomodulatory consequences of oral administration of Lactobacillus rhamnosus strain GG in healthy volunteers. J Dairy Res 2003; 70(2): 165-173.
• 18. Montalto M, Maggiano N, Ricci R, et al. Lactobacillus acidophilus protects tight junctions from aspirin damage in HT-29 cells. Digestion 2004; 69(4): 225-228.
• 19. Galdeano CM, de Moreno de LeBlanc A, Vinderola G, et al. Mechanisms of immunomodulation induced by probiotic bacteria. Clin Vaccine Immunol 2007; 14(5): 485-492.
• 20. McFarland LV. Meta-analysis of probiotics for the prevention of traveler's diarrhea. Travel Med Infect Dis 2007; 5(2): 97-105.
• 21. Resta-Lenert S, Barrett KE. Live probiotics protect intestinal epithelial cells from the effects of infection with enteroinvasive Escherichia coli (EIEC). Gut 2003; 52(7): 988-997.
• 22. Parassol N, Freitas M, Thoreux K, et al. Lactobacillus casei DN-114 001 inhibits the increase in paracellular permeability of enteropathogenic Escherichia coli-infected T84 cells. Res Microbiol 2005; 156(2): 256-262.
• 23. Sartor RB. Cytokines in intestinal inflammation: pathophysiological and clinical considerations. Gastroenterology 1994; 106(2): 533-539.
• 24. Scully P, McKernan DP, Keohane J, et al. Plasma cytokine profiles in females with irritable bowel syndrome and extra-intestinal co-morbidity. Am J Gastroenterol 2010; 105(10): 2235-2243.
• 25. Nataro JP, Guerrant RL. Chronic consequences on human health induced by microbial pathogens: Growth faltering among children in developing countries. Vaccine 2017; 35: 6807-6812.
• 26. Azad MAK, Sarker M, Wan D. Immunomodulatory Effects of Probiotics on Cytokine Profiles. Biomed Res Int 2018; 2018: 8063647.
• 27. Rojas-Cartagena C, Flores I, Appleyard CB. Role of tumor necrosis factor receptors in an animal model of acute colitis. Cytokine 2005; 32(2): 85-93.
• 28. Sainathan SK, Hanna EM, Gong Q, et al. Granulocyte macrophage colony-stimulating factor ameliorates DSS-induced experimental colitis. Inflamm Bowel Dis 2008; 14(1): 88-99.
• 29. Jeengar MK, Thummuri D, Magnusson M, et al. Uridine Ameliorates Dextran Sulfate Sodium (DSS)-Induced Colitis in Mice. Sci Rep 2017; 7(1) :3924.