title: Shenghua Decoction reduces uterine bleeding and regulates T-cell paradigm in human deciduas of RU486 medical abortion.
abstract: BACKGROUND
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Key Laboratory for Tumor Immunology and Traditional Chinese Medicine Immunology, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory for Rare & Uncommon Disease, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Key Laboratory of Biotechnology Drugs of the Ministry of Public Health, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China; Laboratory for Immunopharmacology of State Administration of Traditional Chinese Medicine, Institute of Basic Medicine, Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.forename: Meng
authors.initials: M
authors.lastname: Zhang
authors.forename: Bin
authors.initials: B
authors.lastname: Wang
authors.forename: Yuzhu
authors.initials: Y
authors.lastname: Li
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Xiangzhong
authors.initials: X
authors.lastname: Zhao
authors.forename: Xianbin
authors.initials: X
authors.lastname: Zhou
authors.forename: Yuqi
authors.initials: Y
authors.lastname: Guo
authors.forename: Guosheng
authors.initials: G
authors.lastname: Jiang
authors.forename: Chengfang
authors.initials: C
authors.lastname: Yao
mesh_terms: D000021:Abortifacient Agents, Steroidal / Q000009:adverse effects; D000028:Abortion, Induced / Q000009:adverse effects*; D000328:Adult; D016207:Cytokines / Q000235:genetics / Q000276:immunology; D003656:Decidua / Q000166:cytology / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D004365:Drugs, Chinese Herbal / Q000494:pharmacology / Q000627:therapeutic use*; D005260:Female; D051858:Forkhead Transcription Factors / Q000378:metabolism; D050990:GATA3 Transcription Factor / Q000378:metabolism; D006801:Humans; D015735:Mifepristone / Q000009:adverse effects; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3 / Q000378:metabolism; D011247:Pregnancy; D020825:T-Box Domain Proteins / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D014592:Uterine Hemorrhage / Q000188:drug therapy* / Q000209:etiology / Q000276:immunology / Q000378:metabolism; D055815:Young Adult; D000098815:T-bet Transcription Factor
chemical_list: D000021:Abortifacient Agents, Steroidal; D016207:Cytokines; D004365:Drugs, Chinese Herbal; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D050990:GATA3 Transcription Factor; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; C000590839:Shenghua Decoction; D020825:T-Box Domain Proteins; D000098815:T-bet Transcription Factor; D015735:Mifepristone
keywords: Cytokine; FPA; GA; GAPDH; GATA binding protein 3; GATA-3; IgG; MACS; MSD; PBS; ROR; RT-PCR; RU486; Reverse transcription-polymerase chain reaction; S.D.; SDS; SHD; Sheng-Hua Decoction; Shenghua Decoction; T-bet; T-box expressed in T cells; T-cell paradigm; T-helper cell; TCM; Th cell; Traditional Chinese Medicine; Transcription factor; Uterine bleeding; frequency of previous abortion; gestational age; glyceraldehyde-3-phosphate dehydrogenase; immunoglobulin G; magnetic affinity cell sorting; mean sac diameter; mifepristone; phosphate buffered saline; retinoic acid-related orphan receptor; sodium dodecyl sulfate; standard deviation