Knockout of MIMP protein in lactobacillus plantarum lost its regulation of intestinal permeability on NCM460 epithelial cells through the zonulin pathway

BMC Gastroenterology

Jianping Wang²  Huanlong Qin¹  Mary Pat Moyer³  Nanqi Huang²  Xingwei Zhang²  Meijin Huang²  Chao Tong²  Chao Li²  Liang Kang²  Zhihua Liu¹ 

[1] Department of Surgery, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai 200233, China;Department of Colorectal Surgery, Gastrointestinal Institute of Sun Yat-Sen University, the Sixth Affiliated Hospital of Sun Yat-Sen University (Guangdong Gastrointestinal Hospital), 26 Yuancun Erheng Road, Guangzhou, Guangdong 510655, China;INCELL Corporation, San Antonio, Texas 78249, USA

关键词: Intestinal permeability;    Zonulin;    Micro integral membrane protein;    Lactobacillus plantarum;   

Others  :  1121796
DOI  :  10.1186/1471-230X-14-171

 received in 2013-10-14, accepted in 2014-09-29,  发布年份 2014

【 摘 要 】

Background

Previous studies indicated that the micro integral membrane protein located within the media place of the integral membrane protein of Lactobacillus plantarum CGMCC 1258 had protective effects against the intestinal epithelial injury. In our study, we mean to establish micro integral membrane protein -knockout Lactobacillus plantarum (LPKM) to investigate the change of its protective effects and verify the role of micro integral membrane protein on protection of normal intestinal barrier function.

Methods

Binding assay and intestinal permeability were performed to verify the protective effects of micro integral membrane protein on intestinal permeability in vitro and in vivo. Molecular mechanism was also determined as the zonulin pathway. Clinical data were also collected for further verification of relationship between zonulin level and postoperative septicemia.

Results

LPKM got decreased inhibition of EPEC adhesion to NCM460 cells. LPKM had lower ability to alleviate the decrease of intestinal permeability induced by enteropathogenic-e.coli, and prevent enteropathogenic-e.coli -induced increase of zonulin expression. Overexpression of zonulin lowered the intestinal permeability regulated by Lactobacillus plantarum. There was a positive correlation between zonulin level and postoperative septicemia. Therefore, micro integral membrane protein could be necessary for the protective effects of Lactobacillus plantarum on intestinal barrier.

Conclusion

MIMP might be a positive factor for Lactobacillus plantarum to protect the intestinal epithelial cells from injury, which could be related to the zonulin pathway.

【 授权许可】

   
2014 Liu et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Butler CC, Duncan D, Hood K: Does taking probiotics routinely with antibiotics prevent antibiotic associated diarrhoea? BMJ 2012, 344:e682.
• [2]Liu Z, Qin H, Yang Z, Xia Y, Liu W, Yang J, Jiang Y, Zhang H, Wang Y, Zheng Q: Randomised clinical trial: the effects of perioperative probiotic treatment on barrier function and post-operative infectious complications in colorectal cancer surgery - a double-blind study. Aliment Pharmacol Ther 2011, 33(1):50-63.
• [3]Sivieri K, Morales ML, Adorno MA, Sakamoto IK, Saad SM, Rossi EA: Lactobacillus acidophilus CRL 1014 improved “gut health” in the SHIME reactor. BMC Gastroenterol 2013, 13:100. BioMed Central Full Text
• [4]Liu ZH, Kang L, Wang JP: Basic and clinical research on the regulation of the intestinal barrier by lactobacillus and its active protein components: a review with experience of one center. Mol Biol Rep 2014. [Epub ahead of print]
• [5]Ahmad H, Berzin TM, Yu HJ, Huang CS, Mishkin DS: Central endoscopy reads in inflammatory bowel disease clinical trials: the role of the imaging core lab. Gastroenterol Rep (Oxf) 2014, 2(3):201-206.
• [6]Roberts LM, McCahon D, Holder R, Wilson S, Hobbs FD: A randomised controlled trial of a probiotic ‘functional food’ in the management of irritable bowel syndrome. BMC Gastroenterol 2013, 13:45. BioMed Central Full Text
• [7]Liu Z, Ma Y, Qin H: Potential prevention and treatment of intestinal barrier dysfunction using active components of lactobacillus. Ann Surg 2011, 254(5):832-833. author reply 833
• [8]Liu Z, Shen T, Moyer MP, Qin H: Identification of the lactobacillus SLP domain that binds gastric mucin. Front Biosci 2012, 17:2128-2143.
• [9]Granato D, Bergonzelli GE, Pridmore RD, Marvin L, Rouvet M, Corthesy-Theulaz IE: Cell surface-associated elongation factor Tu mediates the attachment of lactobacillus johnsonii NCC533 (La1) to human intestinal cells and mucins. Infect Immun 2004, 72(4):2160-2169.
• [10]Horie M, Ishiyama A, Fujihira-Ueki Y, Sillanpaa J, Korhonen TK, Toba T: Inhibition of the adherence of escherichia coli strains to basement membrane by lactobacillus crispatus expressing an S-layer. J Appl Microbiol 2002, 92(3):396-403.
• [11]Johnson-Henry K, Wallace JL, Basappa NS, Soni R, Wu GK, Sherman PM: Inhibition of attaching and effacing lesion formation following enteropathogenic escherichia coli and Shiga toxin-producing E. Coli infection. Infect Immun 2001, 69(11):7152-7158.
• [12]Johnson-Henry KC, Donato KA, Shen-Tu G, Gordanpour M, Sherman PM: Lactobacillus rhamnosus strain GG prevents enterohemorrhagic escherichia coli O157:H7-induced changes in epithelial barrier function. Infect Immun 2008, 76(4):1340-1348.
• [13]Yan F, Polk DB: Probiotic bacterium prevents cytokine-induced apoptosis in intestinal epithelial cells. J Biol Chem 2002, 277(52):50959-50965.
• [14]Yan F, Cao H, Cover TL, Whitehead R, Washington MK, Polk DB: Soluble proteins produced by probiotic bacteria regulate intestinal epithelial cell survival and growth. Gastroenterology 2007, 132(2):562-575.
• [15]Liu Z, Ma Y, Yang J, Zhang P, Moyer MP, Qin H: Expression of the lactobacillus plantarum surface layer MIMP protein protected NCM460 epithelial cells from enteroinvasive escherichia coli infection. Cell Physiol Biochem 2011, 27(1):99-108.
• [16]Liu Z, Shen T, Chen H, Zhou Y, Zhang P, Ma Y, Moyer MP, Zhang M, Chu Z, Qin H: Functional characterization of MIMP for its adhesion to the intestinal epithelium. Front Biosci 2012, 17:2106-2127.
• [17]Liu Z, Ma Y, Moyer MP, Zhang P, Shi C, Qin H: Involvement of the mannose receptor and p38 mitogen-activated protein kinase signaling pathway of the microdomain of the integral membrane protein after enteropathogenic escherichia coli infection. Infect Immun 2012, 80(4):1343-1350.
• [18]Johnson-Henry KC, Hagen KE, Gordonpour M, Tompkins TA, Sherman PM: Surface-layer protein extracts from lactobacillus helveticus inhibit enterohaemorrhagic escherichia coli O157:H7 adhesion to epithelial cells. Cell Microbiol 2007, 9(2):356-367.
• [19]Tripathi A, Lammers KM, Goldblum S, Shea-Donohue T, Netzel-Arnett S, Buzza MS, Antalis TM, Vogel SN, Zhao A, Yang S, Arrietta MC, Meddings JB, Fasano A: Identification of human zonulin, a physiological modulator of tight junctions, as prehaptoglobin-2. Proc Natl Acad Sci U S A 2009, 106(39):16799-16804.
• [20]Thomas KE, Sapone A, Fasano A, Vogel SN: Gliadin stimulation of murine macrophage inflammatory gene expression and intestinal permeability are MyD88-dependent: role of the innate immune response in celiac disease. J Immunol 2006, 176(4):2512-2521.
• [21]Fasano A: Zonulin and its regulation of intestinal barrier function: the biological door to inflammation, autoimmunity, and cancer. Physiol Rev 2011, 91(1):151-175.
• [22]Duerksen DR, Wilhelm-Boyles C, Veitch R, Kryszak D, Parry DM: A comparison of antibody testing, permeability testing, and zonulin levels with small-bowel biopsy in celiac disease patients on a gluten-free diet. Dig Dis Sci 2010, 55(4):1026-1031.
• [23]Qin H, Zhang Z, Hang X, Jiang Y: L. Plantarum prevents enteroinvasive escherichia coli-induced tight junction proteins changes in intestinal epithelial cells. BMC Microbiol 2009, 9:63. BioMed Central Full Text
• [24]Liu ZH, Ma YL, Shen TY, Chen HQ, Zhou YK, Zhang P, Zhang M, Chu ZX, Qin HL: Identification of DC-SIGN as the receptor during the interaction of lactobacillus plantarum CGMCC 1258 and dendritic cells. World J Microbiol Biotechnol 2011, 27(3):603-611.
• [25]Liu Z, Zhang P, Ma Y, Chen H, Zhou Y, Zhang M, Chu Z, Qin H: Lactobacillus plantarum prevents the development of colitis in IL-10-deficient mouse by reducing the intestinal permeability. Mol Biol Rep 2011, 38(2):1353-1361.
• [26]Liu ZH, Shen TY, Zhang P, Ma YL, Moyer MP, Qin HL: Protective effects of lactobacillus plantarum against epithelial barrier dysfunction of human colon cell line NCM460. World J Gastroenterol 2010, 16(45):5759-5765.
• [27]Liu Z, Shen T, Zhang P, Ma Y, Qin H: Lactobacillus plantarum surface layer adhesive protein protects intestinal epithelial cells against tight junction injury induced by enteropathogenic escherichia coli. Mol Biol Rep 2011, 38(5):3471-3480.
• [28]Liu ZH, Huang MJ, Zhang XW, Wang L, Huang NQ, Peng H, Lan P, Peng JS, Yang Z, Xia Y, Liu WJ, Yang J, Qin HL, Wang JP: The effects of perioperative probiotic treatment on serum zonulin concentration and subsequent postoperative infectious complications after colorectal cancer surgery: a double-center and double-blind randomized clinical trial. Am J Clin Nutr 2013, 97(1):1-10.
• [29]Pfeiler EA, Klaenhammer TR: Role of transporter proteins in bile tolerance of lactobacillus acidophilus. Appl Environ Microbiol 2009, 75(18):6013-6016.
• [30]Russell WM, Klaenhammer TR: Efficient system for directed integration into the lactobacillus acidophilus and lactobacillus gasseri chromosomes via homologous recombination. Appl Environ Microbiol 2001, 67(9):4361-4364.
• [31]Mohamadzadeh M, Pfeiler EA, Brown JB, Zadeh M, Gramarossa M, Managlia E, Bere P, Sarraj B, Khan MW, Pakanati KC, Ansari MJ, O'Flaherty S, Barrett T, Klaenhammer TR: Regulation of induced colonic inflammation by lactobacillus acidophilus deficient in lipoteichoic acid. Proc Natl Acad Sci U S A 2011, 108(Suppl 1):4623-4630.
• [32]Arrieta MC, Madsen K, Doyle J, Meddings J: Reducing small intestinal permeability attenuates colitis in the IL10 gene-deficient mouse. Gut 2009, 58(1):41-48.
• [33]Huang W, Shang WL, Wang HD, Wu WW, Hou SX: Sirt1 overexpression protects murine osteoblasts against TNF-alpha-induced injury in vitro by suppressing the NF-kappaB signaling pathway. Acta Pharmacol Sin 2012, 33(5):668-674.
• [34]Navarro-Rodriguez T, Silva FM, Barbuti RC, Mattar R, Moraes-Filho JP, de Oliveira MN, Bogsan CS, Chinzon D, Eisig JN: Association of a probiotic to a helicobacter pylori eradication regimen does not increase efficacy or decreases the adverse effects of the treatment: a prospective, randomized, double-blind, placebo-controlled study. BMC Gastroenterol 2013, 13:56. BioMed Central Full Text
• [35]Sapone A, de Magistris L, Pietzak M, Clemente MG, Tripathi A, Cucca F, Lampis R, Kryszak D, Carteni M, Generoso M, Iafusco D, Prisco F, Laghi F, Riegler G, Carratu R, Counts D, Fasano A: Zonulin upregulation is associated with increased gut permeability in subjects with type 1 diabetes and their relatives. Diabetes 2006, 55(5):1443-1449.
• [36]Jakava-Viljanen M, Palva A: Isolation of surface (S) layer protein carrying Lactobacillus species from porcine intestine and faeces and characterization of their adhesion properties to different host tissues. Vet Microbiol 2007, 124(3–4):264-273.
• [37]Zhou Y, Qin H, Zhang M, Shen T, Chen H, Ma Y, Chu Z, Zhang P, Liu Z: Lactobacillus plantarum inhibits intestinal epithelial barrier dysfunction induced by unconjugated bilirubin. Br J Nutr 2010, 104(3):390-401.
• [38]Wang W, Uzzau S, Goldblum SE, Fasano A: Human zonulin, a potential modulator of intestinal tight junctions. J Cell Sci 2000, 113(Pt 24):4435-4440.
• [39]Liu Z, Shi C, Yang J, Zhang P, Ma Y, Wang F, Qin H: Molecular regulation of the intestinal epithelial barrier: implication in human diseases. Front Biosci 2011, 16:2903-2909.
• [40]Liu Z, Shi C, Yang J, Zhang P, Ma Y, Wang F, Qin H: Molecular regulation of the intestinal epithelial barrier: implication in human diseases. Front Biosci 2012, 17:2903-2909.
• [41]Jayashree B, Bibin YS, Prabhu D, Shanthirani CS, Gokulakrishnan K, Lakshmi BS, Mohan V, Balasubramanyam M: Increased circulatory levels of lipopolysaccharide (LPS) and zonulin signify novel biomarkers of proinflammation in patients with type 2 diabetes. Mol Cell Biochem 2014, 388(1–2):203-210.
• [42]Chen X, Lamont JT: Overview of clostridium difficile infection: implications for China. Gastroenterol Rep (Oxf) 2013, 1(3):153-158.
• [43]Lalles JP: Intestinal alkaline phosphatase: multiple biological roles in maintenance of intestinal homeostasis and modulation by diet. Nutr Rev 2010, 68(6):323-332.
• [44]Bates JM, Akerlund J, Mittge E, Guillemin K: Intestinal alkaline phosphatase detoxifies lipopolysaccharide and prevents inflammation in zebrafish in response to the gut microbiota. Cell Host Microbe 2007, 2(6):371-382.
• [45]Alpers DH, Zhang Y, Ahnen DJ: Synthesis and parallel secretion of rat intestinal alkaline phosphatase and a surfactant-like particle protein. Am J Physiol 1995, 268(6 Pt 1):E1205-E1214.
• [46]Sheth P, Samak G, Shull JA, Seth A, Rao R: Protein phosphatase 2A plays a role in hydrogen peroxide-induced disruption of tight junctions in Caco-2 cell monolayers. Biochem J 2009, 421(1):59-70.
• [47]Marasa BS, Xiao L, Rao JN, Zou T, Liu L, Wang J, Bellavance E, Turner DJ, Wang JY: Induced TRPC1 expression increases protein phosphatase 2A sensitizing intestinal epithelial cells to apoptosis through inhibition of NF-kappaB activation. Am J Physiol Cell Physiol 2008, 294(5):C1277-C1287.