| Objective:Folic acid is an essential nutrient which involved in the synthesis of DNA,but also involved in a range of other enzymatic reactions,including protein synthesis and metabolism of vitamin.Folate deficiency not only affects the mother’s health,but also inhibit the normal development of embryonic stem cells.So folate deficiency has an important relationship with the fetus’ development.Therefore,the mother needs to add extra folic acid to meet the growth and development of the fetus during pregnancy.In addition,some studies have found that certain genetic changes which caused by folate deficiency were linked to the occurrence and development of intestinal lesions(Crohn’s disease,irritable bowel syndrome,colorectal cancer).The purpose of this experiment is to initially explore whether folic acid dificiency will affect their offsprings’ gut function and development or not,when we give them without folic acid food feeding before two months of pregnancy and perinatal maternal production period(Subjects are SD rats).We have known that the Wnt/b-catenin and CDX2 which are two important pathway proteins that related to the function and development of intestinal barrier.So we want to learn whether folic acid dificiency will affect their offsprings’ gut function and development or not by studying the expression levels of two pathways protein(Wnt/β-catenin and CDX2).All these we do will providing a more comprehensive understanding to further understand and diagnosis clinical symptoms with diarrhea of children.Methods:In this study,subjects were SD rats.Two months before pregnancy,SD adult females were given standard diets with folic acid or experimental diets without folic acid,were used as control groups and experimental groups.Two months later,the two kinds of female groups were mated to conceive with the adult males(all were feed with standard diets)respectively.We singled out 20 pregnant females from the experimental group and control group to be produced respectively.When the age of pups grow up to 7-10 days,a number of experimental group and control group pups were killed to be observed the development of the overall situation of the gut and the organizations would be taken to prepare for the subsequent experimental analysis.Parts of tissues were frozen in-80℃ degrees freezer for the protein and RNA extraction,the other parts were fixed with 4%formaldehyde and embedded in paraffin for the HE staining and immunehistochemical study.Results:Visually observed,no matter the small intestine or the large intestine have no morphological abnormalities.However,under the microscope we have seen adverse development throughout the intestine with HE staining and corresponding morphological changes have been seen in the intestine crypts and villus enterocytes.Compared with the control,the immunohistochemistry showed that the expression of lysozyme which is the small intestinal tissue-specific expression products of Paneth cells,declined,while the changes is not obvious in the large intestine.In addition,one kind of mucin(MUC2)which is the representative of goblet cells also decreasedMeanwhile we found that folic acid deficiency affected the intestinal barrier function.One of the most important intestinal barrier is the mechanical barrier which consisted by tight junction proteins between cells.We have learned that ZO-1/Claudin-1 were two representative proteins of tight junction proteins between intestinal cells in SD rats.In this study,we found that the folate deficiency group significantly decreased the expression of the ZO-1/Claudin-1 both on the nucleic acid and the protein levels compared with the control group.Conclusions:Congenital folate deficiency has a certain effect on the intestinal barrier development,from the experimental results,we have found that the structure of the intestinal villus and crypts and the intestinal barrier molecules all effected by congenital folate deficiency. |
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