Rotavirus, fermented milk CBAL74, not fermented cow?milk. RV contamination induces alterations of the cytoskeleton through ERK pathway activation via phosphorylating actin cross-linking/bundling proteins19. RV contamination were assessed: epithelial barrier damage (tight-junction proteins and transepithelial electrical resistance evaluation), and inflammation (reactive oxygen species, pro-inflammatory cytokines IL-6, IL-8 and TNF-, and mitogen-activated protein kinase pathway activation). Pre-incubation with FM-CBA L74 resulted in an inhibition of epithelial barrier damage and inflammation mediated by mitogen-activated protein kinase pathway activation?induced by RV infection. Modulating several protective mechanisms, the postbiotic FM-CBAL74 exerted a preventive action against RV contamination. This approach could be a disrupting nutritional strategy against one of the most common killers for the pediatric age. (RV), a segmented double-stranded RNA virus of the?CBAL74 (FM-CBAL74) is among the best characterized Solenopsin and studied postbiotics in the pediatric age11C16. Results from two randomized-controlled trials suggested that FM-CBAL74 can prevent AGE in young children11,12. A positive regulation of several defense mechanisms, including the modulation of gut barrier, the stimulation of adaptative (secretory immunoglobulin A, sIgA), and of innate immunity (human alpha-defensins 1C3; human beta-defensin 2; cathelicidin LL-37) has been exhibited in experimental models and in clinical trials11C14. In addition, the dietary supplementation with FM-CBAL74 has been associated with a beneficial modulation of gut microbiome structure and function in neonates and young children15,16. Prevention is the key for controlling RV-induced AGE. A well-characterized cellular model of RV contamination Solenopsin provided the opportunity to investigate the protective action of FM-CBAL74 against RV-induced AGE. We found that FM-CBAL74 was able to efficiently prevent all main aspects of RV contamination with a significant impact on cellular damage and inflammatory response, through the downregulation of mitogen-activated protein (MAP) kinase pathway. Altogether, these data suggest the potential of the postbiotic approach based on the use of FM-CBAL74 in preventing RV-induced AGE. This approach could be a disrupting nutritional strategy against one of the most common killers for the pediatric age. Results Cellular damage The RV infectivity is commonly demonstrated by the immunofluorescence staining of viral capsid protein VP6 in human enterocytes17,18. The RV contamination of Caco-2 cells was confirmed by an increase of VP6 protein quantification and mRNA levels (non-infected cells (NI); #p? ?0.05 RV-infected cells; p? ?0.05 NFM?+?RV. Rotavirus, fermented milk CBAL74, not fermented cow?milk. RV contamination induces alterations of the cytoskeleton through ERK pathway activation via phosphorylating actin cross-linking/bundling proteins19. This mechanism induces a rearrangement of F-actin filaments20. As showed in Fig.?1, panel B, non-infected cells showed a regular distribution of cytoskeleton actin filaments. In contrast, RV-infected enterocytes showed a marked alteration of cytoskeleton Solenopsin structure with a disorganization of F-actin filaments. Pre-incubation with FM-CBAL74, but not with non-fermented cows milk (NFM), guarded the cells against RV-induced rearrangements of F-actin filaments (Fig.?1, panel B). RV contamination induces apoptosis in human enterocytes21,22. An increase in necrotic cells (positive only for propidium, PI) and late apoptotic cells (positive for both PI and Annexin V) confirmed the pro-apoptotic effect induced by RV contamination (Rotavirus, fermented milk CBAL74, not fermented cow?milk. These results provided evidence on ERK/JNK pathway involvement in the protective action of FM-CBAL74 against RV contamination. Intestinal permeability RAD21 To investigate the protective action of FM-CBAL74 against gut barrier alteration induced by RV contamination, we evaluated the transepithelial electric resistance (TEER), Solenopsin the expression of the tight junction (TJ) proteins, occludin and zonula occludens -1 (ZO-1), and of the cell adhesion molecule E-cadherin in Caco-2 cells monolayer. RV contamination determined a significant decrease of transepithelial resistance (TEER) in human enterocytes (Rotavirus, fermented milk CBAL74, not fermented cow?milk. These results showed that FM-CBAL74 was able to prevent RV induced alteration of gut barrier integrity.?In Supplementary Information 3, we provide a?3D video?reconstruction from Z-stack?acquisition of Occludin.? Inflammatory response Oxidative stress and swelling are related pathophysiological events in infectious diseases25 carefully. To help expand investigate the protecting actions of FM-CBAL74 on inflammatory response induced by RV disease, we looked into oxidative tension (ROS creation) and pro-inflammatory cytokines (IL-6, IL-8 and TNF-) response in human being enterocytes. As demonstrated in Fig.?4, -panel A, RV increased ROS production. The pretreatment with FM-CBAL74, however, not with NFM, inhibited the RV-induced ROS boost (noninfected cells (NI); #p? ?0.05 RV-infected cells; p? ?0.05 NFM?+?RV. Rotavirus, fermented dairy.
Rotavirus, fermented milk CBAL74, not fermented cow?milk
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