The Effect Of Kidney-replenishing Herb On The Indoleamine 2,3-dioxygenase Of Maternal-fetal Interface And The Balance Of Th Cytokine

Section one: The Relationship of abortion and the Expression of Indoleamine 2, 3-dioxygenase(ID0) on Villus and TrophoblastObjective: To study the relationship of abortion and the expression of indoleamine 2,3- dioxygenase (IDO) on villus and trophoblast. Methods: RT-PCR was applied to analyze the mRNA transcription of IDO on villus of normal pregnancy and inevitable abortion and JAR cells; immunohistochemistry was applied to analyze the expression of IDO protein on villus; Western blot was applied to determinate the expression of IDO protein on trophoblast cultured in vitro; high-performance liquid chromatography was applied to determinate whether there is kynurenine in cell culture supernates of trophoblast.Results: The expression of IDO mRNA and protein on villus of inevitable abortion is lower than that of normal pregnancy; IDO mRNA do not express on JAR cells; IDO protein expresses on trophoblast cultured in vitro; and there is kynurenine in cell culture supernates of trophoblast.. Conclusion: Appropriate expression of IDO on villus is necessary for normal pregnancy; IDO protein expresses on the trophoblast and is activited.Section two: The Effect of Kidney-replenishing Herb on the Indoleamine 2, 3-dioxygenase of Human Trophoblast Cultured in Vitro and the balance of Th cytokineObjective: To study the effect of Kidney-replenishing herb on the indoleamine 2, 3-dioxygenase(ID0) of human trophoblast cultured in vitro and the balance of Th cytokines. Methods: Trophoblasts was cultured in vitro for 24,48,72 hours,Group1: with control serum, without IDO inhibitor, 1-methyl-tryptophan (1-MT).Group2: with control serum, with 1-MT 500μmol/L.Group3: with control serum, with 1-MT 1000μmol/LGroup4: with serum made from Kidney-replenishing herb, without 1-MT.Group5: with serum made from Kidney-replenishing herb, with 1-MT 500μmol/L.Group6:with serum made from Kidney-replenishing herb, with 1-MT 1000μmol/L.RT-PCR was applied to analyze the IDOmRNA transcription of trophoblast;Western blot was applied to determinate the expression of IDO protein on trophoblast. Trophoblast and decidual lymphocytes were cocultured and the groups were the same as above, ELISA was applied to determinate the concentration of IL-10 and IFN-γ in coculture supernates; high-performance liquid chromatography was applied to determinate the kynurenine and tryptophan concentration in cell culture supernates, the ratio of kyn/try was used to determinate IDO activity. Results: IDOmRNA and protein can be detected in human trophoblast cultured in vitro; the inhibitor of IDO make the balance of Th cytokine depart from Th2; the serum made from herbal medicine improve the expression of IDOmRNA and protein, promote IDO activity and make the balance of Th cytokine depart from Th1.Conclusion: Kidney-replenishing herb improve the expression of IDOmRNA and protein, promote IDO activity to appropriate value, resume the balance of Th cytokine and regulate the materno-fetal tolerance .Section three: The Effect of Kidney-replenishing Herb on the Indoleamine 2,3-dioxygenase of Decidual Macrophages and Th cytokineObjective: To study the effect of Kidney-replenishing herb on theindoleamine 2,3-dioxygenase(ID0) of decidual macrophages and Thcytokine.Methods: RT-PCR was applied to analyze the IDOmRNA transcription ofdecidue; decidual macrophages was cultured in vitro for 24,48,72 hours,Group1: with control serum, without IDO inhibitor, 1-methyl-tryptophan (1-MT).Group2: with control serum, with 1-MT 500μmol/L.Group3: with control serum, with 1-MT 1000μmol/LGroup4: with serum made from Kidney-replenishing herb, without 1-MT.Group5: with serum made from Kidney-replenishing herb, with 1-MT 500μmol/L.Group6:with serum made from Kidney-replenishing herb, with 1-MT 1000μmol/L.Western blot was applied to analyze the expression of IDO protein on decidual macrophages. Decidual macrophages and lymphocytes were cocultured and the groups were the same as above, ELISA was applied to determinate the IL-10 and IFN-γ concentration in coculture supernates; high-performance liquid chromatography was applied to determinate the kynurenine and tryptophan concentration in cell culture supernates, the ratio of kyn/try was used to determinate IDO activity. Results: The decidual IDOmRNA transcription of normal pregnancy is higher than that of inevitable abortion; the inhibitor of IDO make the balance of Th cytokine depart from Th2; the serum made from herbal medicine improve the expression of IDO protein and IDO activity, and resume the balance of Th cytokine.Conclusion: Appropriate IDOmRNA transcription and IDO activity of decidue is necessary for normal pregnancy; Kidney-replenishing herb improve IDO activity to appropriate value.