The mediating role of gut microbiota in the associations of prenatal maternal combined exposure to lead and stress with neurodevelopmental deficits in young rats.

Prenatal single and combined exposure to lead (Pb) and stress (Ps) impairs neurodevelopment. Prenatal single exposure to Pb or Ps affects the composition of intestinal microbiota, and bidirectional communication between gut microbiota and central nervous system has been well recognized. However, whether gut microbiota mediated the effects of prenatal Pb+Ps co-exposure on neurodevelopmental deficits remains unclear. This study established rat models with prenatal single and combined exposure to Ps and Pb. We investigated the effects of such prenatal single and combined exposure on hippocampal structures using morphological analyses, on learning/memory using the Morris-water-maze test, and on fecal microbiota using 16S rRNA sequencing. The mediating roles of gut microbiota were analyzed using the bootstrap method. The study found both single and combined exposure affected hippocampal ultra-structures and spatial learning/memory, and the most significant impairments were observed in the Pb+Ps group. Prenatal Pb+Ps co-exposure decreased fecal microbial alpha/beta-diversity. Significantly lower levels of B/F-ratio, class-Bacteroidia, order-Bacteroidales, and family-S24-7, and significantly higher levels of class-Bacilli, order-Lactobacillales, family-Lactobacillaceae, and genus-Lactobacillus were observed in the co-exposure group, compared with the controls. Increased relative abundances of genus-Helicobacter mediated the detrimental effect of prenatal Ps+Pb co-exposure on learning/memory [β (95%CI) for the total and indirect effects: - 10.70 (-19.19, -2.21) and - 4.65(-11.07, -1.85)], accounting for 43.47% of the total effect. As a result, increased relative abundances of genus-Lactobacillus alleviated the adverse effects of the co-exposure on learning/memory, and the alleviation effect accounted for 44.55% of the direct effect [β (95%CI) for the direct and indirect effects: - 0.28(-0.48, -0.08) and 0.13(0.01, 0.41)]. This study suggested that prenatal combined exposure to Pb and Ps induced more impairments in offspring gut microbiota and neurodevelopment than single exposure, and alterations in fecal microbiome may mediate the developmental neurotoxicity induced by such prenatal co-exposure.