In vitro anti-inflammatory activity among probiotic Lactobacillus species isolated from fermented foods
Introduction
Due to the enormous changes in environmental conditions and industrial revolution, modern man has adopted western lifestyle leading to psycho-emotional stress, inflammatory response and chronic infections (Bosma-den Boer, van Wetten, & Pruimboom, 2012). The intake of high calorie foods, low physical activity had affected the immune system of host and increased the mortality rate (Bosma-den Boer et al., 2012). It was reported that, the probiotic microbes would reduce the side-effects emerging from the unavoidable modern life style, by restoring the gut dysbiosis (Yoo & Kim, 2016). The Indian traditional foods have a plethora of probiotic microbes, anti-oxidant activity, nutritive properties, dietary fibres, etc., and hence are recognized as “functional foods” (Sarkar, Lohith Kumar, Dhumal, Panigrahi, & Choudhary, 2015). Diverse species of Lactobacillus were identified in fermented foods of North-East India (NEI) with potential health promoting benefits (Devi et al., 2016, Tamang et al., 2015). Probiotic microbes used in food products as starter cultures get adhered to the intestinal epithelial cells and play a significant role in the host immunomodulation (Neish, 2009, Son et al., 2005, Swain et al., 2014). The interaction of potential probiotic strain to host epithelial cells would enhance their probiotic properties and is considered as an important aspect in the current scenario (Behnsen et al., 2013, van Hermert et al., 2010). Many strains of Lactobacillus and Bifidobacterium have conferred their health benefits by adhering to the epithelial layer, release of pro- and anti-inflammatory cytokines (CKs) (Ramiah, van Reenen, & Dicks, 2007), excluding pathogens, etc. (Wang et al., 2014). Moreover, the release of CKs bestows host-microbe interaction (Foligne et al., 2015). Such beneficial microbes also aid in minimizing the adverse symptoms during diarrhoea, inflammatory bowel disease (IBD), allergic reactions, physio-emotional stress, bacterial or viral infections, peptic ulcers and Chron's disease (Nishiyama et al., 2016, Son et al., 2005). The T-helper cells like Th1 and Th2 stimulate inflammatory CKs and provide protection against invading pathogenic microbes, inflammatory response, tissue destruction in the host (Plaza-Diaz, Gomez-Llorente, Fontana, & Gil, 2014). Studies on CKs like Interleukin (IL)-6, IL-8 IL-10, IL-12 and tumor necrosis factor (TNF)-α have shown to stimulate pro-inflammatory response triggering fever, inflammation, disturb cell barrier function, etc. (Lee et al., 2016). The anti-inflammatory CKs like IL-4, IL-10, transforming growth factor (TGF) etc. are found to raise their levels under inflammatory conditions (Presti et al., 2015). However, the mechanism of probiotic action during immunomodulation is highly strain-specific and has a direct co-relation towards the health aspect of host (Duary, Batish, & Grover, 2014). A host under oxidative stress, inflammation and during invasion of pathogenic microbes is found to stimulate immune cells and activate the inflammatory CKs leading to disturbed cell barrier function, increased symptoms of infection and intestinal injury (Kim et al., 2015, Paszti-Gere et al., 2012). The current study is focussed on the selection of a potential probiotic strain with better anti-inflammatory action through quantitative Real Time PCR assay.
It was reported that the beneficial microbes possess surface layer binding proteins like mucin binding protein (Mub) with adhesion domains (MucBP) that aid in colonization to epithelial cells of host (Devi and Halami, 2017, Devi et al., 2015, Etzold et al., 2014). However, the application of bioinformatics tools on the domain repeats would elucidate the adhesion mechanism, co-aggregation of related bacteria and colonization to the gut (Nishiyama et al., 2016, Rychli et al., 2017). Till date, the evolutionary aspects of the adhesion domain repeats between probiotic and pathogenic microbes were not elucidated and an attempt was made in the current study.
Section snippets
Bacterial isolates and culture conditions
Potential probiotic cultures Lactobacillus plantarum subsp. plantarum MTCC 5422 (isolated from fermented cereal) and CFR MFT1 (MCC 3446) (isolated from Eup), Lactobacillus paraplantarum MTCC 9483 (isolated from gundruk) and Lactobacillus rhamnosus GG or ATCC 53,103 (a proven probiotic strain) were used in the present study (Devi et al., 2016). All cultures were grown in MRS broth (deMan, Ragosa, and Sharpe, HiMedia, Mumbai, India) for 12–16 h at 37 °C. Subsequently, the pathogenic strain such
Adhesion of Lactobacillus strains to Caco-2 cells
In our present study, adhesion assay and SEM analysis to Caco-2 cells was evaluated for four Lactobacillus strains, i.e., L. plantarum subsp. plantarum MTCC 5422 and MCC 3446, L. paraplantarum MTCC 9483 and L. rhamnosus GG (a standard probiotic strain). Fig. 1 shows the adherence of probiotic cultures to gastrointestinal cell line, and it was observed that all the four strains possessed good adherence ability. An adhesion score of 58, 46, 128 and 62% was observed for MCC 3446, MTCC 5422, MTCC
Discussion
Recent studies have established that the functional aspects of probiotics are highly strain-specific and their biological effects cannot be judged at species level (Ramos, Thorsen, Schwan, & Jespersen, 2013). This makes the researchers to screen continuously for novel probiotic strains with more promising health benefits.
Adherence of probiotic bacteria to host intestinal layer is considered as a principal criterion to exhibit functional properties (Duary et al., 2014, Kaushik et al., 2009).
Conclusion
The current study elucidated the selection of potent probiotic Lactobacillus strain from vegetable origin with anti-inflammatory activity. Among all the strains used, Lactobacillus paraplantarum MTCC 9483 was found to possess strong anti-inflammatory property during oxidative stress, LPS-induction, and pathogen invasion conditions, with alleviated levels for IL-4, IL-10 and TLR-2. In addition, a study on in silico analysis suggested that the MucBP domain (D10) might have evolved from Listeria
Conflict of interest
The authors claim no conflicts of interest
Ethical approval
This article does not contain any studies on either animals models or humans participants.
Acknowledgements
The authors thank Director (CSIR-CFTRI) for the constant encouragement. We thank the Head, Department of Biochemistry for providing the facilities for Cell-culture and Real-time PCR assays. SMD extend her sincere gratitude to SERB- Department of Science and Technology, New Delhi for endorsing the study under Start-up grant scheme (Project No. SB/YS/LS-353/2013).
References (59)
- et al.
Interleukin 4 acts as an inducer of decay-accelerating factor gene expression in human intestinal epithelial cells
Gastroenterology
(1996) - et al.
Interaction of probiotic Lactobacillus and Bifidobacterium strains with human intestinal epithelial cells: adhesion properties, competition against enteropathogens and modulation of IL-8 production
International Journal of Food Microbiology
(2008) - et al.
Discrimination and divergence among Lactobacillus plantarum-group (LPG) isolates with reference to their probiotic functionalities from vegetable origin
Systematic and Applied Microbiology
(2016) - et al.
Diversity and evolutionary aspects of mucin binding (MucBP) domain repeats among Lactobacillus plantarum group strains through comparative genetic analysis
Systematic and Applied Microbiology
(2017) - et al.
Immunomodulation properties of multi-species fermented milks
Food Microbiology
(2016) - et al.
Scanning electron microscopy of target cells and molecular weight determination of a bacteriocin produced by Lactococcus lactis D53
International Journal of Food Microbiology
(1994) Microbes in gastrointestinal health and disease
Gastroenterology
(2009)- et al.
Expression of the mucus adhesion genes Mub and MapA, adhesion-like factor EF-Tu and bacteriocin gene plaA of Lactobacillus plantarum 423, monitored with real-time PCR
International Journal of Food Microbiology
(2007) - et al.
Strain-specific probiotics properties of Lactobacillus fermentum, Lactobacillus plantarum and Lactobacillus brevis isolates from Brazilian food products
Food Microbiology
(2013) - et al.
Traditional and ayuvedic foods of Indian origin
Journal of Ethnic Foods
(2015)
Vascular endothelial growth factor (VEGF) in colonic epithelial cells in induced by hypoxia and reduced by Tumor necrosis factorα (TNFα)
Gastroenterology
Histidine inhibits oxidative stress- and TNF-α-induced interleukin-8 secretion in intestinal epithelial cells
FEBS Letters
Immunomodulatory and cytotoxic effects of various Lactococcus strains on the murine macrophage cell line J774.1
International Journal of Food Microbiology
Antibiotic resistance in Lactococcus species from bovine milk: Presence of a mutated multidrug transporter mdt(A) gene in susceptible Lactococcus garvieae strains
Veterinary Microbiology
Screening of adhesive lactobacilli with antagonistic activity against Campylobacter jejuni
Food Control
Induction of cytokine formation by human intestinal bacteria in gut epithelial cell lines
Journal of Applied Microbiology
Combined Effect of 25-Hydroxycholesterol and IL-1β on IL-8 Production in Human Colon Carcinoma Cell Line (Caco-2)
Inflammation
Probiotics: Properties, examples, and specific applications
Cold Spring Harbour Perspectives in Medicine
Toll-like receptor 2 Arg677Trp polymorphism is associated with susceptibility to tuberculosis in tunisian patients
Clinical and Vaccine Immunology
Listeria monocytogenes surface proteins: From genome predictions to function
Microbiology and Molecular Biology Reviews
Chronic inflammatory diseases are stimulated by current life style: How diet, stress levels and medication prevent our body from recovering
Nutrition & Metabolism
In vitro adhesion and anti-inflammatory properties of native Lactobacillus fermentum and Lactobacillus delbrueckii spp.
Journal of Applied Microbiology
Regulation of the anti-inflammatory cytokines interleukin-4 and interleukin-10 during pregnancy
Frontiers in Immunology
Screening, characterization and in vitro evaluation of probiotic properties among lactic acid bacteria through comparative analysis
Probiotics and Antimicrobial Proteins
Immunomodulatory activity of two potential probiotic strains in LPS-stimulated HT-29 cells
Genes & Nutrition
Assessing the adhesion of putative indigenous probiotic lactobacilli to human colonic epithelial cells
Indian Journal of Medical Research
Structural basis for adaptation of lactobacilli to gastrointestinal mucus
Environmental Microbiology
Correlation between in vitro and in vivo immunomodulatory properties of lactic acid bacteria
World Journal of Gastroenterology
Clostridium butyricum activates TLR2-mediated MyD88-independent signaling pathway in HT-29 cells
Molecular and Cellular Biochemistry
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