Shenghua Decoction reduces uterine bleeding and regulates T-cell paradigm in human deciduas of RU486 medical abortion.
Excessive uterine bleeding is the most common and problematic side effect of RU486 medical abortion. Shenghua Decoction (SHD) is a well-known traditional Chinese herbal prescription for reducing uterine bleeding induced by RU486 medical abortion. However, its therapeutic mechanism still remains unclear. The Th1/Th2/Th17/Treg paradigm plays an important role in achieving maternal-fetal immunotolerance and its bias participates in RU486-induced abortion. Our previous research on mice demonstrated that the uterine bleeding volume is negatively related to the proportions of Th1 and Th17 cells whereas positively related to the proportions of Th2 and Treg cells. Additionally, Th1-type cytokine inducing effect was identified in our previous study. Therefore, it was hypothesized that SHD reduced the uterine bleeding in RU486 medical abortion by inducing Th1/Th2/Th17/Treg paradigm bias. The purpose of this study was to determine the regulatory effect and the mechanism of SHD on human decidual Th1/Th2/Th17/Treg paradigm for alleviating uterine bleeding in RU486 medical abortion.
90 women within seven weeks of a normal intrauterine pregnancy, who elected for termination of pregnancy, were divided into three groups; vacuum aspiration group, RU486 group, and SHD-RU486 group. Duration of uterine bleeding was recorded and volume of uterine bleeding was measured by the method of alkaline hematin photometric. To determine the regulatory effect of SHD on Th1/Th2/Th17/Treg paradigm, the proportions of Th1/Th2/Th17/Treg cells in the decidua of different groups were analyzed using a FACS calibur. Correlation was analyzed in order to demonstrate the relationship between the Th1/Th2/Th17/Treg paradigm and the uterine bleeding in RU486 medical abortion. Moreover, to elucidate the mechanism underlying the T-cell paradigm regulating of SHD, the mRNA and protein expressions of subset-specific transcription factors (T-bet, GATA-3, RORγt, and Foxp3) for the differentiation of Th1/Th2/Th17/Treg paradigm in human decidual CD4(+) T cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) assay and western blot analysis respectively. Moreover, the mRNA expression of the characteristic cytokines of Th1/Th2/Th17/Treg paradigm (IFNγ, IL-4, IL-17A, TGF-β) were analyzed by RT-PCR assay.
Compared with RU486 group, both the uterine bleeding volume and duration reduced significantly in SHD-RU486 group. Both the duration and the volume of the uterine bleeding demonstrated negative correlation with the proportions of Th1 and Th17 cells, whereas showed positive correlation with Th2 and Treg cells. SHD increased the proportions of Th1 and Th17 cells whereas decreased those of Th2 and Treg cells. Thus, the ratios of Th1/Th2 and Th17/Treg cells elevated markedly after SHD treatment. SHD promoted the mRNA as well as the protein expressions of subset-specific transcription factors for the differentiation of Th1 and Th17 subsets (T-bet and RORγt) while inhibited those of Th2 and Treg cells (GATA-3 and Foxp3). Moreover, the mRNA expression of Th1- and Th17- type cytokines (IFNγ and IL-17A) was up-regulated while that of Th2-type and Treg-produced cytokines (IL-4 and TGF-β) was down-regulated significantly after SHD administration.
Th1/Th2/Th17/Treg paradigm bias was involved in RU486 medical abortion. SHD reduced the uterine bleeding efficiently by inducing Th1 and Th17 skews in the maternal-fetal of RU486 medical abortion patients. The regulatory effect of SHD on Th1/Th2/Th17/Treg paradigm in RU486 medical abortion is attributed to the modulation of transcription and protein expression of subset-specific transcription factors for T-cell subsets differentiation and their characteristic cytokines.
Li X
，Zhang M
，Wang B
，Li Y
，Wang L
，Zhao X
，Zhou X
，Guo Y
，Jiang G
，Yao C
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The dynamic changes of T-bet(+)/GATA-3(+) and RORgammat(+)/FOXP3(+) cells in recipient spleens and grafts after rat orthotopic liver transplantation.
Rejection of transplanted liver occurs when the host generates alloantigen-reactive T cells, and CD4(+) T-cell subsets, including Th1, Th2, T17 and iTreg, could be involved in changing the dynamics of graft rejection onset. In the current immunosuppressive strategies, rejection is treated as an undifferentiated process that is uniform, which results in the failure of tolerance induction. Here, we established a rejection model to observe the reciprocal interaction of CD4(+) T-cell subsets in the complex networks of the immune system.
Orthotopic liver transplantation (OLT) from male inbred Lewis rats (n=15) to male inbred Brown Norway (BN) rats was performed by Kamada's two-cuff technique without reconstruction of the hepatic artery. OLT from BN to BN rats (n=5) was performed as a control. Recipients were sacrificed on postoperative days 3, 5, 7, 9, 11, 13 and 15. Recipient spleens and grafts were harvested, fixed in 10% neutral formalin, and embedded in paraffin. Meanwhile, hematoxylin and eosin and immunohistochemical staining was done, acute rejection was graded by the Banff scheme, and the number of T-bet(+), GATA-3(+), RORgammat(+) and FOXP3(+) cells in the spleen and grafts were counted.
In recipient spleens, the T-bet(+) and RORgammat(+) cells were increased more significantly in the mild acute rejection (AR) group than in the control group (P=0.016, P=0.009, respectively), while both cell types were decreased in the moderate AR group. Compared with the control group, the RORgammat(+) cells did not differ significantly in the severe AR group, while the T-bet(+) cells were significantly decreased (P=0.465, P=0.009, respectively). The GATA-3(+) cells were significantly decreased in the mild AR group compared with the control group (P=0.009). With regard to the FOXP3(+) cells, there was no significant difference between the control and mild AR groups (P=0.754), while they were significantly decreased in the moderate and severe AR groups (P=0.028, P=0.009, respectively). The ratio of T-bet(+)/GATA-3(+) cells was more associated with AR than was the RORgammat(+)/FOXP3(+) cell ratio in the early stage. In the graft, the T-bet(+) and RORgammat(+) cells were significantly increased in mild, moderate and severe AR groups compared with the control group (P<0.009). The expression of GATA-3(+) cell was not significantly increased in the mild AR group compared with the control group, while it was significantly increased in the moderate and severe AR groups (P=0.028, P=0.009, respectively). Concerning the FOXP3(+) cells, no significant difference was found between the control and mild AR groups (P=0.347), while they were significantly increased in the moderate and severe AR groups (P<0.009).
Our study revealed the dynamic changes of T-bet(+), GATA-3(+), RORgammat(+) and FOXP3(+) cells in the spleen and grafts of recipient rats. It seems that the ratio of T-bet(+)/GATA-3(+) cells was more associated with AR than was RORgammat(+)/FOXP3(+) cells in the early stage of rejection, while the ratio of RORgammat(+)/FOXP3(+) cells was associated more with the later but more persistent stage of rejection. This study could make a contribution to the optimal selection of immunosuppressive regimens according to the dynamic changes in CD4(+) T-cell subsets at different stages of rejection.
Xu M
，Tan C
，Zhou J
，Huang X
，Dai Z
，Zhu H
，Zhao Y
，Gu F
，Zhou S
，Fan J
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Expression of Transcriptional Factors of T Helper Differentiation (T-bet, GATA-3, RORγt, and FOXP3), MIF Receptors (CD44, CD74, CXCR2, 4, 7), and Th1, Th2, and Th17 Cytokines in PBMC from Control Subjects and Rheumatoid Arthritis Patients.
The macrophage migration inhibitory factor (MIF) plays a pivotal role in the development of rheumatoid arthritis (RA). Previous research indicates that MIF can trigger the expression of cytokine profiles associated with Th1, Th2, and Th17 responses in peripheral blood mononuclear cells (PBMC) from both RA patients and control subjects (CS). Despite these, few studies to date precisely elucidate the molecular mechanisms involved. The present study aimed to associate the expression of Th differentiation TF (T-bet, GATA-3, RORγt) with MIF receptors (CD44, CD74, CXCR2, 4, 7) and Th1, Th2, and Th17 cytokines in PBMC from CS and RA patients.
PBMC from both groups was cultured for 24 h. The expression of the canonical and non-canonical MIF receptors and the TF was determined by flow cytometry. Additionally, multiplex bead analysis was employed to assess the levels of cytokines in the culture supernatants. The findings revealed that T CD4+ lymphocytes in the CS group exhibited a heightened expression of CD74 (p<0.05), whereas RA patients displayed an elevated expression of CXCR7 (p<0.001). Furthermore, T CD4+ lymphocytes from RA patients exhibited greater expression of GATA3, RORγt, and FOXP3, along with elevated levels of pro-inflammatory cytokines compared to the CS group (p<0.001).
These results indicate that CD74 is more prominently expressed in PBMC from the CS group, whereas CXCR7 is more expressed in PBMC from RA patients.
We also noted an increased secretion of Th17 profile cytokines in RA, potentially influenced by the activation of FOXP3 via CD74 and RORγt through CXCR7 using the endocytic pathway.
Zerpa-Hernández DA
，García-Chagollán M
，Sánchez-Zuno GA
，García-Arellano S
，Hernández-Bello J
，Hernández-Palma LA
，Cerpa-Cruz S
，Martinez-Bonilla G
，Nicoletti F
，Muñoz-Valle JF
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ResearchGate
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