Elsevier

Journal of Functional Foods

Volume 47, August 2018, Pages 19-27
Journal of Functional Foods

In vitro anti-inflammatory activity among probiotic Lactobacillus species isolated from fermented foods

https://doi.org/10.1016/j.jff.2018.05.036Get rights and content

Highlights

  • Probiotic L. paraplantarum MTCC 9483 adhered more to epithelial Caco-2 cells.

  • Gene expression of anti-inflammatory cytokines was observed during probiotic treatment.

  • The adhesion domain of Lactobacillus species might have evolved from Listeria species.

  • Isolate MTCC 9483 effectively modulated induced conditions by its anti-inflammatory activity.

Abstract

Probiotic bacteria have the ability to alter immune response conferring treatment to inflammatory diseases. In the current study, probiotic Lactobacillus strains were subjected to adhesion to epithelial Caco-2 cells. Subsequently, gene expression of pro-and anti-inflammatory genes were studied under three stimulation conditions (which includes LPS-challenge, stress induction and pathogens invasion). Herein, immunomodulatory activity of native Lactobacillus strains (n = 3) were compared to L. rhamnosus GG. The studied strains had down-regulated the gene expression of pro-inflammatory cytokines (CKs) (like TNF-α, IL-1α, IL-1β, IL-6 and IL-8) and up-regulated the anti-inflammatory CKs like TGFβ1, IL-4, and IL-10. Gene expression of TLR2 has also been noted. Strain-specific interactions were noticed during the different probiotic treatment. Further, phylogenetic dendrogram of mucin adhesion domain suggested the interaction of Lactobacillus and Listeria species, explaining competitive colonization and exclusion mechanism in an ecological niche. Overall, isolate L. paraplantarum MTCC 9483 has effectively modulated induced conditions by its anti-inflammatory activity.

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).

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