三黄三仙汤治疗再生障碍性贫血的作用机制研究OA

Objective:To explore the effect of Sanhuang Sanxian decoction in the treatment of aplastic anaemia(AA)and its underlying molecular mechanisms in depth.Methods:Splenic lymphocytes were isolated and purified to obtain Naive CD4+T cells,which were then induced to differentiate interleukin-17(IL-17)producing helper T cells(Th17).The immunophenotype of CD4+T cells was analyzed by flow cytometry,and their morphological changes were assessed by optical microscopy.The CCK8 assay detected cell viability after treatment with different concentrations of Sanhuang Sanxian decoction-containing serum.Flow cytometry was used to identify regulatory T cells(Treg)and Th17 cell differentiation in each group.An enzyme-linked immunosorbent assay was used to detect the levels of interleukin(IL)-17A,IL-10 and IL-21 in cells.qPCR was used to detect IL-17F,T-bet mRNA expression in cells.Western blot was performed to detect the protein expression of peroxisome proliferator-activated receptor γ(PPARγ),retinoid-related orphan receptor gamma t(RORγt),and transcription factor forkhead box protein 3(Foxp3)in cells.Results:CCK8 results indicated 10%drug-containing serum as the optimal serum concentration.Compared with the model group,there was a remarkable decreased proportion of Th17 cells in the Sanhuang Sanxian decoction-containing serum group and the Icariin group,whereas there was a significantly increase of the proportion of Treg cells.IL-17A and IL-21 content dramatically decreased and IL-10 content dramatically increased.The expression levels of IL-17F,T-bet mRNA was significantly reduced.The PPARγ and Foxp3 protein expression was markedly elevated,and the RORγt protein expression was markedly reduced.The opposite trend was observed with the addition of the PPARγ inhibitor T0070907.Conclusion:Sanhuang Sanxian decoction was able to effectively alleviated AA,and its mechanism of action may be related to the activation of PPARγ,causing dysregulation of the Foxp3/RORγt pathway.