| Endogenous antimicrobial peptides are an important component in innate immunity of organism and the defensins are a big family in endogenous antimicrobial peptides.β-defensins are important component of defensins family.β-defensins are proved to be a component of antimicrobial barrier and they distribute prominently in muscosal epithelial cells of mammalians and aves.There haven't relating research about defensins of sheep in female reprocuctive tract at present. There also havn't report about the relationship of estrogen and progesterone with defensins in female reprocuctive tract .In this study,we study the expression of beta-defensins of Mongolian sheep in female reprocuctive tract,and the relationship of expression of beta-defensins in female reprocuctive tract with estrogen and progesterone.1 We have demonstratedβ–defensin genes is expressed in the epithelial layers of the vagina,cervix,uterus,and fallopian tubes in the female reproductive tract of Mongolian sheep with RT-PCR. Furthermore,there are different expressions ofβ–defensin in different tissue of female reproductive tract.2 We detected the mRNA ofβ–defensins in the fallopian tubes organs by in situ hybridization. The result show, theβ–defensin mRNA localized in the cytoplast free end of epithelial cell of the fallopian tubes.There aren't expression in proper layer, muscular layer and connective tissue.3 We obtained the primary and passage epithelial cells of the fallopian tubes of mongolian sheep by 0.25% trypsin.The resulting of cultured cells were identified as epithelial cell by morphological technique and immunohistochemical method.The cell healthy were identified by Karyotype analysis. Therefore,the cells we cultured can be applyed to study the relationgship of the sBD-1 expression of epithelial cell of the fallopian tubes and hormone.4 For explored the relationgship of the expression of sBD-1 in the female reproductive tract with estrogen and progesterone,we added estrogen and progesterone into the cultured epithelial cell of the fallopian tubes.The passaged cell were cultured for 4 days following treated with serum free medium for 10 hours and then were treated with hormone.The expression of sBD-1 in cultured passaged cell were detected by real-time PCR.The results show ,(1) there is significant difference between blank group and treated groups with 10-8 M,10-9 M ,10-10 M 17-β-Estradiol.However, there is no difference between blank group and treated with 10-8 M 17-β-Estradiol.When treated with 17-β-Estradiol, different cultured time can not give different difference on the expression of sBD-1.Some concentration 17-β-Estradiol can promote the expression of sBD-1 in cultured epithelia cell of the fallopian tubes.(2) There is difference between blank group and treated groups with 10-6 M and 10-7 M progesterone. There is significant difference between blank group and treated groups with 10-8 M and 10-9M progesterone. There is no difference between blank group and treated group with 10-11 M progesterone. Some concentration progesterone can promote the expression of sBD-1 in cultured epithelia cell of the fallopian tubes.(3) There is significant difference between blank group and oestrum simulated group(added 10-8 M 17-β-Estradiol, 10-9Mprogesterone) on the expression of sBD-1. There is significant difference between blank group and gestation simulated group(added 10-10M 17-β-Estradiol, 10-7 M progesterone) on the expression of sBD-1.So, 17-β-Estradiol and progesterone can promote the expression of sBD-1 in cultured epithelia cell of the fallopian tubes.The expression ofβ–defensin in the female reproductive tract have the relationship with the menstrual cycle.This result provided evidence that estrogen and progesterone was one of the factor that can promote the expression of sBD-1,but not the essential factor of expression of sBD-1.5 To determine the mechanism by which estrogen and progesterone promote the expression of sBD-1 in female reproductive tract,cultured epithelia cells were treated with mifiprestone and tamoxifen following treated with estrogen and progesterone corresponding.Then the expression of sBD-1 were dteced by real-time PCR.The results show,there isn't obviously effect when the cell were treated with 10-6 M tamoxifen for 8 hours following treated with 10-8M estrogen.There isn't obviously effect when the cell were treated with 10-6 M mifiprestone for 8 hours following treated with 10-8M progesterone. Estrogen and progesterone promote the expression of sBD-1 by combining with corresponding receptor. |
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