Based on Th17/Treg imbalance to explore the mechanism of Bushen Gutai mixture in the treatment of recurrent spontaneous abortion
Objective Based on the imbalance of helper T cell 17(Th17)/regulatory T cell(Treg),the possible mechanism of Bushen Gutai Mixture in the treatment of recurrent abortion(RSA)was investigated.Methods After 1 week of adaptive feeding,80 SD rats(30 male and 50 female)were combined with cages,and the mark after successful pregnancy was detected as day 0 of gestation,and were randomly divided into 5 groups with 10 rats in each group,including blank control group,model group,dydrogesterone group,low-dose Bushen Gutai mixture group and high-dose Bushen Gutai mixture group.In the afternoon of the 1st to 9th day of gestation,except the blank control group,the other groups were intragaxed with 450 mg·kg-1·d-1hydroxyurea tablet aqueous solution,and intragaxed with 4 mg·kg-1·d-1 mifepristone aqueous solution in the morning of the 10th day of gestation.From day 1 to day 14 of gestation,the blank control group and model group were given equal volume normal saline in the morning,the dydrogesterone group was given 3.02 mg·kg 1·d-1 dydrogesterone aqueous solution,and the low dose group and high dose group were given 9.9 g·kg-1·d-1and 39.6 g·kg-1·d-1 water decoction of Bushen Gutai mixture,respectively.The changes of body mass at 14 days gestation were observed.The spleen index of pregnant mice was calculated.The embryonic growth and development of pregnant mice in each group were observed.He-matoxylin-eosin(HE)staining was performed on decidua tissue and spleen tissue of pregnant mice,and the pathological changes were observed.The contents of interleukin-17(IL-17),tumor necrosis factor-α(TNF-α),interleukin-10(IL-10)and transfor-ming growth factor-β1(TGF-β1)in serum of pregnant mice were detected by ELISA kit.The expression of retinoid-related orphan nuclear receptor γt(RORγt)and forked head spiral transcription factor 3(FOXP3)in decidual tissue was detected by immunofluores-cence staining(IF).The expressions of IL-17,TNF-α,IL-10 and TGF-β1 in decidua of pregnant mice were detected by Western blot.Results Compared with the blank control group,the mass of the 14th celestial body in the model group was significantly decreased(P＜0.01).Spleen index was significantly decreased(P＜0.01).The size of embryo development of pregnant mice was not uni-form,and some embryos showed blood stasis and necrosis.Embryo loss rate was significantly increased(P＜0.01).The decidual tis-sue cells were disordered,with nucleus shrinkage,partial disappearance,cytoplasmic edema,and partial vacuole-like change.The splenic tissue structure was seriously abnormal,some splenic nodules were disordered,and the boundary between red pulp and white pulp was chaotic and blurred.The expression levels of IL-17,TNF-α,and RORγt in serum and decidua tissue were significantly increased(P＜0.01),while the expression levels of IL-10,TGF-β1,and RORγt/FOXP3 were significantly decreased(P＜0.01).Com-pared with model group,the spleen index in dydrogesterone group and Bushen Gutai mixture high-dose group was significantly increased(P＜0.01);Compared with the model group,the embryo size and development of pregnant mice were significantly improved,and only some embryos were absorbed.The rate of embryo loss was significantly decreased(P＜0.01).The arrangement of decidua cells was relatively orderly,the cell morphology was regular,and only a few cells were edema and necrosis.The parenchyma distribution of spleen tissue was more uniform,and the boundary of red pulp and white pulp was clear.The expression levels of IL-17,TNF-α and RORγt in serum and decidua tissue,and the ratios of IL-17/TGF-β1,TNF-α/IL-10 and RORγt/FOXP3 were significantly decreased(P＜0.05,P＜0.01).The expressions of IL-10,TGF-β1 and FOXP3 were significantly increased(P＜0.05,P＜0.01).There was no significant difference in the mass of the 14th object in dydrogesterone group(P＞0.05).The mass of the 14th celestial body in Bushen Gutai mixture high dose group was significantly increased(P＜0.05).Conclusion Bushen Gutai mixture may maintain Th17/Treg immune balance by up-regulating the expression of IL-10,TGF-β1 and FOXP3,and down-regulating the expression of IL-17,TNF-α and RORγt,thereby reducing the embryo loss rate of pregnant mice in RSA model.
Recurrent spontaneous abortionBushen Gutai mixtureImmune imbalanceTh17/TregSD rat
NETL
NSTL
万方数据
