COPD is a prevalent lung disease with significant impacts on public health. Affected airways exhibit pulmonary neutrophilia and consequent secretion of pro-inflammatory cytokines and proteases, which result in lung emphysema. Probiotics act as nonspecific modulators of the innate immune system that improve several inflammatory responses. To investigate the effect of Lactobacillus rhamnosus (Lr) on cigarette smoke (CS)-induced COPD C57Bl/6 mice were treated with Lr during the week before COPD induction and three times/week until euthanasia. For in vitro assays, murine bronchial epithelial cells as well as human bronchial epithelial cells exposed to cigarette smoke extract during 24 hours were treated with Lr 1 hour before CSE addition. Lr treatment attenuated the inflammatory response both in the airways and lung parenchyma, reducing inflammatory cells infiltration and the production of proinflammatory cytokines and chemokines. Also, Lr-treated mice presented with lower metalloproteases in lung tissue and lung remodeling. In parallel to the reduction in the expression of TLR2, TLR4, TLR9, STAT3, and NF-κB in lung tissue, Lr increased the levels of IL-10 as well as SOCS3 and TIMP1/2, indicating the induction of an anti-inflammatory environment. Similarly, murine bronchial epithelial cells as well as human bronchial epithelial cells (BEAS) exposed to CSE produced pro-inflammatory cytokines and chemokines, which were inhibited by Lr treatment in association with the production of anti-inflammatory molecules. Moreover, the presence of Lr also modulated the expression of COPD-associated transcription found into BALF of COPD mice group, i.e., Lr downregulated expression of NF-κB and STAT3, and inversely upregulated increased expression of SOCS3. Thus, our findings indicate that Lr modulates the balance between pro-and anti-inflammatory cytokines in human bronchial epithelial cells upon CS exposure and it can be a useful tool to improve the lung inflammatory response associated with COPD.

IntroductionChronic pulmonary obstructive disease (COPD) is defined as an abnormal inflammatory response of the lungs to noxious particles or gases, mainly cigarette smoke. Thus, new therapeutic approaches are of unquestionable relevance. Without a specific treatment for COPD patients the use of probiotics via supplementation of diet can be a promising target. This project aims to investigate the beneficial effects of Bifidobacterium breve (Bb) and Lactobacillus rhamnosus (Lr) probiotics in lung inflammatory process in mice in a model of lung emphysema.MethodsThis study was approved by the Ethics Committee of the UNIFESP. The COPD was induced by cigarette smoke inhalation of 14 cigarette per day, twice a day, 7 times/week during 60 days in C57Bl/6 mice. The mice were treated with Lr and Bb at the same time. The pro‐inflammatory mediators as IL‐6, TNF, IL‐1 β, CXCL1, CXCL8, CXCL10, KC, CXCL9, CXCL11 and anti‐inflammatory as IL‐10 in bronchoalveolar lavage fluid (BALF) were measured by ELISA. The expression of mRNA of the MMP9 and MMP12, NF‐κB, STAT3 and TLR 2,4 and 9 in lung were analyzed by quantitative RT‐PCR. The NF‐κB was also analyzed by immunolocalization. The lung tissue was used for histological and morphometric analyzes.ResultsBb and Lr attenuated the cellularity in BALF and reduced the pro‐inflammatory cytokines and inversely increased the anti‐inflammatory. Also, the probiotics reduced the expression of MMP9 and 12, NF‐κB, STAT3 and TLR 2,4 and 9 in lung. Then, the probiotic also changed the airway remodeling (inflammatory infiltrate, alveolar enlargement, collagen, and elastic fibers).ConclusionThis study points out that the probiotics decreased the cellularity on the BALF and the expression of the main transcription factors as a consequence they also reduced pro‐inflammatory cytokines and inversely increase anti‐inflammatory. These results contribute to the understanding of Bb and Lr modulator activities.Support or Funding InformationFinancial Support: Foundation for Research Support of the State of São Paulo (FAPESP)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

AbstractCOPD is a prevalent lung disease with significant impacts on public health. Affected airways exhibit pulmonary neutrophilia and consequent secretion of pro-inflammatory cytokines and proteases, which result in lung emphysema. Probiotics act as nonspecific modulators of the innate immune system that improve several inflammatory responses. To investigate the effect of Lactobacillus rhamnosus (Lr) on cigarette smoke (CS)-induced COPD C57Bl/6 mice were treated with Lr during the week before COPD induction and three times/week until euthanasia. For in vitro assays, murine bronchial epithelial cells as well as human bronchial epithelial cells exposed to cigarette smoke extract during 24 hours were treated with Lr 1 hour before CSE addition. Lr treatment attenuated the inflammatory response both in the airways and lung parenchyma, reducing neutrophilic infiltration and the production of pro-inflammatory cytokines and chemokines. Also, Lr-treated mice presented with lower metalloproteases in lung tissue and lung remodeling. In parallel to the reduction in the expression of TLR2, TLR4, TLR9, STAT3, and NF-κB in lung tissue, Lr increased the levels of IL-10 as well as SOCS3 and TIMP1/2, indicating the induction of an anti-inflammatory environment. Similarly, murine bronchial epithelial cells as well as human bronchial epithelial cells (BEAS) exposed to CSE produced pro-inflammatory cytokines and chemokines, which were inhibited by Lr treatment in association with the production of anti-inflammatory molecules. Moreover, the presence of Lr also modulated the expression of COPD-associated transcription found into BALF of COPD mice group, i.e., Lr downregulated expression of NF-κB and STAT3, and inversely upregulated increased expression of SOCS3. Thus, our findings indicate that Lr modulates the balance between pro- and anti-inflammatory cytokines in human bronchial epithelial cells upon CS exposure and it can be a useful tool to improve the lung inflammatory response associated with COPD.

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