Interference Of Neonicotinoids On Neural Differentiation Of Mesenchymal Stem Cells

Neonicotinoids have gradually become the most widely used insecticides in the world.Recently,it has been found that neonicotinoids not only acted on non-target insects such as bees,but also posed toxic effects on vertebrates.The results of animal experiments showed that exposure to neonicotinoids had adverse effects on the nervous system of mice and rats,but the toxicity mechanism and molecular toxicological targets were still unclear.In the present study,human bone marrow mesenchymal stem cells(hBMSCs)were used as an in vitro model to study the interference of three neonicotinoids on the neuronal differentiation of hBMSCs.In addition,the gene expression profiles of hBMSCs exposed to neonicotinoids were analyzed to explore the toxic mechanism and sensitive targets of neonicotinoids.The main results are as follows:The morphology of neurogenic differentiation was analyzed at the cell level,and the expression of neuronal differentiation markers at the gene and protein level was detected at the molecular level.The results showed that IMI and ACE significantly inhibited the length of neurite formed by differentiated cells.IMI significantly inhibited the expression of NESTIN and NF-M protein.In the concentration of 25 ?g/m L and 50 ?g/m L,NIT significantly inhibited the expression of NESTIN and NF-M protein but significantly increased the expression of MAP2 protein.ACE significantly enhanced the expression of NESTIN and MAP2 protein at the concentration of 25 ?g/m L.NIT and ACE significantly inhibited the expression of NESTIN at all exposure concentrations.NIT and ACE significantly inhibited the expression of WISP1,which was the downstream target gene of Wnt/?-catenin signaling pathway.The results showed that the three neonicotinoids interfered with the neuronal differentiation of hBMSCs.Gene chip was used to explore the developmental neurotoxicity mechanism of NEOs and to screen potential sensitive targets.328 differentially expressed genes(DEGs)were induced by IMI,NIT and ACE,accounting for 63.7% of all DEGs.Alcoholism,PI3k-Akt and m TOR signaling pathway related to nervous system were enriched through further analysis of gene expression profiles,also with biological processes related to neuronal cell behavior,learning and memory.NGF,BDNF and SERPINB were screened as the key genes of neonicotinoids.In addition,the results of gene expression profile suggested the role of Wnt/?-catenin signaling pathway in neurogenic differentiation of hBMSCs interfered by neonicotinoids.In the present study,we found that exposure to neonicotinoids interfered with the neuronal differentiation of hBMSCs,and identified a series of targets of neonicotinoids on hBMSCs by transcriptomic data analysis.The results provided a novel evidence for further study of potential toxic effects of neonicotinoids exposure on human body,and scientific data for environmental and health risk assessment and regulation of neonicotinoids.