Investigation of Intestinal Enzyme Activity and Effects of Non-starch Polysaccharide on it

  • Authors

    • Masoumeh Piryaei
    • Anahita Motamedi
    • Atefeh Mehrabi Far
    https://doi.org/10.14419/ijet.v7i4.7.20372

    Received date: September 27, 2018

    Accepted date: September 27, 2018

    Published date: September 27, 2018

  • Cell Extracts, Laboratory Conditions, Intrinsic Immunity, Lipopolysaccharide, Shigella.

  • Abstract

    Background and Purpose: Shigella is a human shigellosis and its lipopolysaccharide is identified by 4TLR. The 4TLR is a family of pseudo-TOLL receptors and many immune routes are triggered by stimulating these receptors. Many studies show increasing of 4TLR expression in Mesenchyme stem cells under the influence of lipopolysaccharide. The main objective of this study was to identify the appropriate lipopolysaccharide of Shigella strains by stimulating the immune system for vaccine studies. Materials and Methods: In this experimental study, the stem cell of human Mesenchymal derived from bone marrow was treated by three dilution of 0.1, 0.01, and 0.001 extract of Shigella strains (Flexneri, Dysentery and Sonnei) containing lipopolysaccharide. Then, the expression of 4TLR at RNA level was evaluated by RT-PCR and Q-PCR techniques. Cells treated with phosphate buffer saline were considered as control group. Findings: The expression of 4TLR was observed in all treatments groups except for treatment groups with relative concentration of 0.001 sonnei and dysentery as well as control group. Changes in 4TLR expression were dose-dependent on all treatment groups. The highest expression was related to the treatment with Shigella Flexneri extract and the smallest was related to Shigella sonnei. The use of pure lipopolysaccharide of Escherichia coli as a positive control showed that the lipopolysaccharide in Shigella extract is responsible for increasing the expression of 4TLR. Conclusion: given the increased expression of 4TLR by Shigella extract, this extract is recommended to increase the efficacy of the vaccine.

  • References

    1. Nhung PH, Ohkusu K, Mishima N, Noda M, Shah MM, Sun X, Phylogeny and species identification of the family Enterobacteri-aceae based on dnaJ sequences. Diagnostic microbiology and infec-tious disease. 2007; 58(2): 153-61.
    2. Key B, Clemens J, Kotloff K. Generic Protocol to Estimate the Burden of Shigella Diarrhoea and Dysenteric Mortal, Tex. Book; 1999.P. 146-51.
    3. 3. Brooks G. Mycoplasmas & cell wall-defective bacteria. U: Brooks GF, Carroll KC, Butel JS. Morsa SA, Mietzner TA, ur. Jawetz, Melnick & Adelberg’s Medical Microbiology. New York: McGraw-Hill Companies, Inc; 2010.
    4. Man AL, Prieto-Garcia ME, Nicoletti C. Improving M cell mediat-ed transport across mucosal barriers: do certain bacteria hold the keys? Immunology. 2004;113(1):15-22.
    5. Trent MS, Stead CM, Tran AX, Hankins JV. Invited review: diver-sity of endotoxin and its impact on pathogenesis. Journal of endo-toxin research. 2006; 12(4):205-23.
    6. 6. Rallabhandi P, Awomoyi A, Thomas KE, Phalipon A, Fujimoto Y, Fukase K, et al. Differential activation of human TLR4 by Esch-erichia coli and Shigella flexneri 2a lipopolysaccharide: combined effects of lipid A acylation state and TLR4 polymorphisms on sig-naling. The Journal of Immunology. 2008; 180(2):1139-47.
    7. 7. Aboussahoud W, Aflatoonian R, Bruce C, Elliott S, Ward J, Newton S, et al. Expression of Toll-like Receptors in Human En-dometrialEpithelial Cells and Cell Lines. Journal of Reproductive Immunology. 2010; 84(1): 41-51.
    8. 8. Aflatoonian R, Tuckerman E, Elliott S, Bruce C, Aflatoonian A, Li T, Menstrual cycledependent changes of Toll-like receptors in endometrium. Human Reproduction. 2007; 22(2): 586-93.
    9. 9. Miyake K, editor. Innate immune sensing of pathogens and dan-ger signals by cell surface Tolllike receptors. Seminars in immunolo-gy; 2007; 19: 3- 10.
    10. 10. Hwa Cho H, Bae YC, Jung JS. Role of Toll- Like Receptors on Human Adipose-Derived Stromal Cells. Stem Cells. 2006;24(12):2744-52.
    11. 11. Akira S, Uematsu S, Takeuchi O. Pathogen recognition and in-nate immunity. Cell. 2006;124(4):783-801.
    12. 12. Tomchuck SL, Zwezdaryk KJ, Coffelt SB, Waterman RS, Danka ES, Scandurro AB. Toll-Like Receptors on Human Mesen-chymal Stem Cells Drive Their Migration and Immunomodulating Responses. Stem Cells. 2008;26(1): 99-107.
    13. DelaRosa O, Lombardo E. Modulation of adult mesenchymal stem cells activity by toll-like receptors: implications on therapeutic po-tential. Mediators of inflammation. 2010;2010.p. 865601-2.
    14. 14. Bäckhed F, Normark S, Schweda EK, Oscarson S, Richter-Dahlfors A. Structural requirements for TLR4-mediated LPS signal-ling: a biological role for LPS modifications. Microbes and infec-tion. 2003; 5(12):1057-63.
    15. Jahantigh D, Saadati M, Ramandi MF, Mousavi M, Zand A. Novel Intranasal Vaccine Delivery System by Chitosan Nanofibrous Membrane Containing N-Terminal Region of Ipad Antigen as a Na-sal Shigellosis Vaccine, Studies in Guinea Pigs. Journal of Drug De-livery Science and Technology. 2014;24(1):33-9.
    16. Saadati M, Heiat M, Nazarian S, Barati B, Honari H, Doroudian M, et al. Cloning and Expression of N-terminal Region of IpaD from Shigella dysenteriae in E. coli. Journal of Paramedical Sciences. 2010;1(4):12-7.
    17. 17. Mallaei F, Saadati M, Honari H, Nazariyan Sh, Eghtedardoust M, Heiat M, et al. Cloning and expression of ipaC gene from Shi-gella dysenteriae. Kowsar Medical Journal 2011; 16: 1-6.
    18. 18. Emadedin M, Aghdami N, Taghiyar L, Fazeli R, Moghadasali R, Jahangir S, et al. Intraarticular injection of autologous mesen-chymal stem cells in six patients with knee osteoarthritis. Archives of Iranian medicine. 2012(15):422-8.
    19. 19. Shi L, Wang J-S, Liu X-M, Hu X-Y, Fang Q. Upregulated func-tional expression of Toll like receptor 4 in mesenchymal stem cells induced by lipopolysaccharide. Chinese medical journal. 2007;120(19):1685-8.
    20. 20. Lin H, Xu R, Zhang Z, Chen L, Shi M, Wang F-S. Implications of the immunoregulatory functions of mesenchymal stem cells in the treatment of human liver diseases. Cellular & molecular immunolo-gy. 2011;8(1):19-22.
    21. 21. Tyndall A. Mesenchymal stem cell treatments in rheumatology [mdash] a glass half full? Nature Reviews Rheumatology. 2014;10(2):117-24.
    22. 22. DelaRosa O, Dalemans W, Lombardo E. Tolllike receptors as modulators of mesenchymal stem cells. Frontiers in immunology. 2012;3:182.
    23. 23. Pevsner-Fischer M, Morad V, Cohen-Sfady M, Rousso-Noori L, Zanin-Zhorov A, Cohen S, et al. Toll-like receptors and their ligands control mesenchymal stem cell functions. Blood. 2007;109(4): 1422-32.
    24. 24. Loppnow H, Brade H, Dürrbaum I, Dinarello CA, Kusumoto S, Rietschel ET, et al. IL-1 induction-capacity of defined lipopolysac-charide partial structures. The Journal of Immunology. 1989; 142(9): 3229-38.
    25. 25. Seydel U, Oikawa M, Fukase K, Kusumoto S, Brandenburg K. Intrinsic conformation of lipid A is responsible for agonistic and an-tagonistic activity. European Journal of Biochemistry. 2000; 267(10): 3032-9.
    26. 26. Hajjar AM, Ernst RK, Tsai JH, Wilson CB, Miller SI. Human Toll-like receptor 4 recognizes host-specific LPS modifications. Na-ture immunology. 2002;3(4):354-9.
    27. 27. Kim HM, Park BS, Kim J-I, Kim SE, Lee J, Oh SC, et al. Crys-tal structure of the TLR4-MD-2 complex with bound endotoxin an-tagonist Eritoran. Cell. 2007;130(5):906-17.
    28. 28. Huber M, Kalis C, Keck S, Jiang Z, Georgel P, Du X, et al. R-form LPS, the master key to the activation ofTLR4/MD-2-positive cells. European journal of immunology. 2006; 36(3): 701-11.
    29. Henricson B, Perera P, Qureshi N, Takayama K, Vogel S. Rhodop-seudomonas sphaeroides lipid A derivatives block in vitro induction of tumor necrosis factor and endotoxin tolerance by smooth lipo-polysaccharide and monophosphoryl lipid A. Infection and immuni-ty. 1992;60(10):4285-90.
    30. Visintin A, Halmen KA, Latz E, Monks BG, Golenbock DT. Phar-macological inhibition of endotoxin responses is achieved by target-ing the TLR4 coreceptor, MD-2. The Journal of Immunology. 2005; 175(10): 6465-72.

  • Downloads

  • How to Cite

    Piryaei, M., Motamedi, A., & Mehrabi Far, A. (2018). Investigation of Intestinal Enzyme Activity and Effects of Non-starch Polysaccharide on it. International Journal of Engineering and Technology, 7(4.7), 23-27. https://doi.org/10.14419/ijet.v7i4.7.20372