Early Life Emulsifier Exposure Induces Dysbiosis And Defective Intestinal Barrier In The Offspring

Background and Aims: The first 3 years of life is a crucial period for the development and maturation of gut microbiota,which is defined as the early stage of life.Properinoculation of intestinal microbiota during early stage of life is of great importance to individuals' health.Disruption of this inoculation process may lead to ascending morbidity of multiple diseases.The main determinants of early life gut microbiota colonizing involve delivery mode,feeding mode and environment exposure.It is proposed that the maternal-infant microbiota transmission may occur through delivery and breast-feeding.It has been revealed that interference with maternal intestinal microbiota during gestation and lactation might impact their offspring's microbiota and this impact could even persist to the third generation.Polysorbate80(P80)is a kind of synthetic amphiphilic nonionic surfactant,which is ubiquitously applied to the manufacture of foods and medications as a kind of emulsifier.So females may inevitably take in P80.Both in vivo and ex vivo studies have revealed that P80 could disrupt intestinal barrier function and induce chronic low-dose inflammation and elicit dysbiosis and metabolic disorders,while whether maternal intake of P80 has an impact on offspring remains unknown.This study aims to investigate whether maternal intake of P80 during gestation and lactation could influence the ontogeny and intestinal microbiota of the second and third generation of offspring.Methods:Six-week-old C57bl/6 mice were randomly separated into 2 groups.One group was provided with sterile water throughout the course of the study while the other group was continuously interfered with 1%P80 dissolved in drinking water.After co-housed with male mice from the same group for a week,successfully pregnant female mice were raised along before the offspring were delivered.The second generation of offspring were co-housed with their mother and fed by maternal breast milk before weaning.Part of the second generation mice in both groups were sacrificed and the body weight and quantity of the liver were documented.The rest of the second generation in both groups were raised alone for 5 weeks with sterile water.After that,each female mouse was co-housed with male mice from the same group for a week,respectively.Successfully pregnant female mice were raised along before the offspring were delivered.The third generation of offspring were co-housed with their mother for 3 weeks before sacrifice.The body weight and quantity of the liver were documented.Fecal samples and intestinal specimens of both the second and third generations were collected and stored for later assays.Ontogeny was estimated via comparing body weight and liver mass and measuring the length of villus using HE stained specimens.Periodic Acid-Schiff(PAS)dye was utilized to count the amount of goblet cells.Immunofluorescent staining was applied to evaluate the expression of secretory Ig A(s Ig A).The expression of components of tight junction and MUC2 were quantified via realtime-PCR(RT-PCR).RT-PCR was also utilized to analyze the expression of inflammatory cytokines.The 16 sr DNA sequencing techniques was applied to analyzing the fecal microbiota.Results:(1)The P80 group of the second generation had shorter intestinal villus and the P80 group of the third generation had shorter jejunal villus.Colonic glands of the P80 group were shorter in both the second and the third generations.(2)The composition of gut microbiota differed between the P80 group and the control group in both the second and the third generations.Mucispirillumwas of lower abundance in both the second and the third generations.(3)The expression of IL-6 and IL-1? in the P80 group was higher in both the second and the third generations.(4)The amount of colonic goblet cells were lower in the P80 group in both the second and the third generations.The expression of ZO-1 and claudin-3 was lower in the P80 group in both the second and the third generations.The expression of MUC2 was lower in the P80 group in both the second and the third generations.The positive rate of s Ig A dye was lower in the P80 group in both the second and the third generations.Conclusion: Maternal P80 exposure may disrupt the offspring's intestinal barrier function,induce dysbiosis and low grade inflammation,and further impact the ontogeny of the gut.