OBJECTIVETo analysis the differential expression of decidua tissue proteins and effective mechanisms of recurrent abortion mice with Shoutai Wan, and explore the mechanism of Shoutai Wan in preventing miscarriage.

METHODThe abortion-prone CBA/J x DBA/2 matings were established as the model of recurrent abortion and the nonabortion-prone CBA/J x BALB/c matings were used as the model of normal pregnancy. The model of recurrent abortion CBA/J x DBA/2 of mice pregnant were randomly divided into four groups according to the sequence of pregnancy, including model group, Shoutai Wan low-dose group, Shoutai Wan middle-dose group and Shoutai Wan high-dose group. From the 1st day of pregnant, mice of normal group, model group, Shoutai Wan low-dose group (3 g x kg x d(-1)), Shoutai Wan middle-dose group (6 g x kg x d(-1)) and Shoutai Wan high-dose group (12 g x kg x d(-1)) are oral administration in different doses. On the 14th day of pregnancy, all mice are killed and the embryo loss rate (ELR) was counted. The expression of differential proteins of mice decidua tissues were separated by means of 2-DE and identified by MALDI-TOF-MS. The functions of identified proteins were further analysed according to bioinformatics resources.

RESULTCompared with model group, low-dose Shoutai Wan can not significantly improve the model of recurrent abortion in pregnant mice ELR; Shoutai Wan middle-dose and high-dose group of pregnant mice ELR were significantly decreased (P < 0.01). The results showed that the well-resolved, reproducible 2-DE patterns of mice decidua tissues of model group, normal group and Shoutai Wan low middle high-dose group were obtained. Through comparative proteome analysis of decidua tissues of all groups, 30 differential expression protein spots which maybe related to recurrent abortion and Shoutai Wan intervention were identified by MALDI-TOF-MS. These differential expression proteins mainly refer to invasion of the blastocyst, blood vessel remodeling and cell apoptosis.

CONCLUSIONShoutai Wan can decrease recurrent abortion mice ELR significantly, and play a role in preventing miscarriage. Recurrent abortion is a complicated process refer to diverse proteins participate. For several protein spots expression of decidua tissues in recurrent abortion mice was regulated by Shoutai Wan, it provides contribution to the effect characteristic of multitarget.

Abortion, Habitual ; metabolism ; pathology ; prevention & control ; Animals ; Decidua ; drug effects ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Mice ; Pregnancy ; Proteome ; metabolism

OBJECTIVE: To observe the expression of progesterone receptor (PR), interleukin-1β (IL-1β), and cyclooxygenase-2 (COX-2) induced by lipopolysaccharide (LPS) or Toll-like receptor 4 antagonist (TLR4 mAb) in decidual cells in vitro, and then to explore the effect of LPS and its antagonist on PR of decidual cells and the relation between PR and inflammatory cytokines. METHODS: We isolated and cultured human decidua of early abortion in the sterile state. When the cells passaged to the 4th generation, the cells were randomly divided into 6 pore plates: A control group was added the culture medium alone; experimental group I was added 100 ng/mL of LPS; experimental group II was add 1 μg/mL of TLR4 mAb; experimental group III was added 3 μg/ mL of TLR4 mAb; experimental group IV was added 1 μg/mL of TLR4 mAb pretreatment for 24 h, and then 100 ng/mL LPS; and experimental group V was added 3 μg/mL of TLR4 mAb pretreatment for 24 h, and then 100 ng/mL LPS for 24 h culture. Subsequently, HE staining and immunofluorescence were used to observe the morphology and identify the purity of decidual cells in the 6 groups. The levels of mRNA expression of PR, IL-1β, and COX-2 were detected by reverse transcription PCR (RT-PCR). RESULTS: LPS reduced the mRNA expression of PR (P<0.05), increased the mRNA expression of IL-1β and COX-2 (P<0.05). TLR4 mAb increased the mRNA expression of PR (P<0.05) and reduced the mRNA expression of IL-1β (P<0.05) after LPS-stimulated decidual cells. High concentrations of TLR4 mAb reduced the mRNA expression of COX-2 (P<0.05) after LPS stimulated decidual cells. CONCLUSION The mRNA expression of PR is reduced, and the mRNA expressions of IL-1β and COX-2 are increased after LPS-stimulated decidual cells in vitro. TLR4 mAb antagonize the role of LPS on PR, IL-1β, and COX-2.
Adult ; Cells, Cultured ; Cyclooxygenase 2 ; genetics ; metabolism ; Decidua ; cytology ; metabolism ; Female ; Humans ; Interleukin-1beta ; genetics ; metabolism ; Lipopolysaccharides ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Receptors, Progesterone ; genetics ; metabolism ; Toll-Like Receptor 4 ; antagonists & inhibitors ; Young Adult

OBJECTIVETo investigate the expression of Rho-GDI in the decidual tissues of patients preeclampsia and explore its clinical implication.

METHODSReal-time PCR, Western blotting and immunohistochemistry were used to detect the mRNA and protein expressions of Rho-GDI in the decidual tissues from 30 normal women with full-term pregnancy, 30 patients with early-onset severe preeclampsia and 30 with late-onset severe preeclampsia.

RESULTSRho-GDI expression was found mainly on the cell membrane and in the cytoplasm and nuclei of the decidual cells, occasionally occurring in the stroma. Both the mRNA and protein expressions of Rho-GDI in the decidual tissues were significantly higher in the normal pregnancy group than in the two severe preeclampsia groups (P<0.05), and the patients with late-onset severe preeclampsia had the lowest expressions of Rho-GDI.

CONCLUSIONThe lowered expression of Rho-GDI in the deciduas might be involved in the pathogenesis and progression of preeclampsia.

Adult ; Decidua ; metabolism ; Female ; Guanine Nucleotide Dissociation Inhibitors ; genetics ; metabolism ; Humans ; Pre-Eclampsia ; metabolism ; Pregnancy ; RNA, Messenger ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; methods ; Young Adult ; rho-Specific Guanine Nucleotide Dissociation Inhibitors

OBJECTIVETo investigate the relationship between the expression of uncoupling protein 2 (UCP2) and ratio of interleukin-10/interferon-gamma (IL10/IFNgamma) in the macrophages from patients with unexplained recurrent spontaneous abortion (URSA).

METHODSTwelve women undergoing selective termination of normal early pregnancy (control) and 11 having URSA were included in this study. Magnetic cell sorting (MACS) was used to isolate the macrophages in the decidua, and the expression of UCP2 was detected with flow cytometry. Cytokine (IL10 and IFNgamma) secretion by the macrophages was detected by enzyme-linked immunosorbent spot-forming (ELISPOT) cell assay.

RESULTSCompared to the control group, the women with URSA showed significantly decreased expression of UCP2 on decidual macrophage (136-/+25 vs 201-/+31, P<0.01), and the expression of UCP2 was positively correlated to the ratio of IL10/IFNgamma(r=0.73, P<0.01).

CONCLUSIONUCP2 may play an important role in the regulation of macrophage activity and cytokine secretion to contribute to spontaneous abortion.

Abortion, Habitual ; metabolism ; Adult ; Cytokines ; metabolism ; Decidua ; cytology ; metabolism ; Female ; Humans ; Interferon-gamma ; secretion ; Interleukin-10 ; secretion ; Ion Channels ; metabolism ; Macrophage Activation ; Mitochondrial Proteins ; metabolism ; Pregnancy ; Uncoupling Protein 2

OBJECTIVETo investigate the expression of annexin V in the decidua tissues of preeclampsia patients and explore its clinical significance.

METHODSReal-time PCR, Western blotting and immunohistochemistry were employed to detect the mRNA and protein expressions of annexin V in the deciduas from 35 normal pregnant women at full term, 38 early onset severe preeclampsia patients and 33 late onset severe preeclampsia patients.

RESULTSAnnexin V was found on the cell membrane and in the cytoplasm of the decidual cells and stroma. Both the mRNA and protein of annexin V expressions in the decidua tissues were significantly different between normal pregnancy group and early or late onset severe preeclampsia group (P<0.05), being the highest in normal pregnancy group and the lowest in early onset severe preeclampsia group.

CONCLUSIONThe low expression of annexin V in the deciduas might participate in the hypercoagulability state in preeclampsia patients.

Adult ; Annexin A5 ; metabolism ; Case-Control Studies ; Decidua ; metabolism ; Female ; Humans ; Immunohistochemistry ; Pre-Eclampsia ; metabolism ; pathology ; Pregnancy ; RNA, Messenger ; genetics

BACKGROUNDBAFF, the B cell activation factor, is a member of the tumor necrosis factor (TNF) ligand family that binds to BCMA, TACI, and BAFF-R. Previous studies have shown that members of the TNF family are detected in human placental trophoblast cells, but the expression patterns of BAFF involved in human decidua and the differential expression of BAFF between normal pregnancy and miscarriage are still incompletely documented or unknown. This study was designed to investigate the expression of BAFF and BAFF-R in the trophoblast and decidua of normal early pregnant women and recurrent spontaneous abortion (RSA) patients.

METHODSForty-five patients with RSA and 45 normal pregnant women were included in this study. By reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemical experiments, we explored the expression of BAFF and BAFF-R in the maternal-fetal interface of normal early pregnant women and RSA patients.

RESULTSAnalysis by RT-PCR and Western blotting revealed that BAFF was detected in both trophoblast and decidua of all the samples, and the expression level was higher in the tissues of normal early pregnant women (P<0.05) than that of recurrent spontaneous abortion patients under the same gestational weeks. Messages for BAFF-R were absent. Immunohistochemical experiments showed that expression of BAFF was cell-specific which was localized to villous cytotrophoblast and syncytiotrophoblast cells in trophoblast and to stromal cells in decidua. Whereas BAFF was prominent on the trophoblast and decidua of normal early pregnant women, it was decreased in the tissues of RSA patients.

CONCLUSIONSBAFF might steer maternal leukocytes away from a harmful immune response and toward a favorable one and play a potentially vital role for successful pregnancy.

Abortion, Habitual ; metabolism ; B-Cell Activating Factor ; analysis ; genetics ; physiology ; Decidua ; chemistry ; metabolism ; Female ; Humans ; Immunohistochemistry ; Interleukin-10 ; genetics ; Pregnancy ; RNA, Messenger ; analysis ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Trophoblasts ; chemistry ; metabolism

Human placental decidua basalis-mesenchymal stem cells (PDB-MSCs) are multipotent cells from the human term placenta, which are ethically conducive, easily accessible and high-yielding source. PDB-MSCs can differentiate into adipogenic, osteogenic and neurogenic cells under appropriate conditions, which may be an attractive and alternative source of seed cells for tissue engineering. To investigate the effect of hypoxia (1% O2) on human PDB-MSCs and the expression of cytokine, PDB-MSCs were isolated from human placenta by density gradient centrifugation and cultured in the Dulbecco's modified Eagle's medium-high glucose (DMEM-HG) containing 10% fetal bovine serum (FBS), and the fifth passage of PDB-MSCs were taken. PDB-MSCs were divided into 4 groups according to the concentrations of O2 and FBS: 20% O2, 10% FBS; 20% O2, 0% FBS; 1% O2, 10% FBS; 1% O2, 0% FBS. The proliferation and apoptosis of PDB-MSCs were detected by MTT and flow cytometric analysis at the time points of 6 h, 12 h, 24 h, 48 h, 72 h, and 96 h, respectively. Vascular endothelial growth factor (VEGF) released from PDB-MSCs was detected by enzyme-linked immunosorbent assay (ELISA) at the same time points. The results showed that hypoxia enhanced the proliferation of PDB-MSCs at 12 h under the condition of 10% FBS, while at 24 h under the condition of 0% FBS (P<0.01, n=3). In normoxia, the cells cultured in 10% FBS displayed a significant proliferation compared to those cultured in 0% FBS. However, in hypoxia, the number of cells cultured in 0% FBS (serum deprivation) increased significantly compared to that cultured in 10% FBS at 24 h and 96 h respectively (P<0.05, P<0.01, n=3). With the flow cytometric analysis of cell apoptosis under the condition of hypoxia and serum deprivation, we found that hypoxia and serum deprivation did not induce PDB-MSCs apoptosis (P>0.05, n=3). This conclusion may relate to the expression of VEGF which needs further research. In conclusion, the results obtained indicate that PDB-MSCs are able to bear hypoxia and serum deprivation, suggesting that PDB-MSCs can be used as seed cells for ischemia related tissue engineering.
Apoptosis ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Decidua ; cytology ; Female ; Humans ; Mesenchymal Stromal Cells ; cytology ; Placenta ; cytology ; Pregnancy ; Tissue Engineering ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo observe the clinical therapeutic effect of acupuncture massage for primary dysmenorrhea and its influence on prostaglandin F2alpha level in menstrual fluid.

METHODSNinety patients with primary dysmenorrhea were assigned equally to the acupuncture group, the massage group and the control group (oral administrated with Somiton tablet). The treatment course was 3 menstrual cycles. The effect in relieving menalgia was evaluated using numerical rating scale (NRS) before and after treatment and change of PGF2alpha content in menstrual fluid was tested as well.

RESULTSThe NRS score was 0.60 +/- 0.67 in the acupuncture group and 0.53 +/- 0.63 in the massage group respectively, significantly lower than that in the control group, which was 2.30 +/- 1.73 (P < 0.01). The PGF2alpha level after treatment was significantly lower in the acupuncture group (176.58 +/- 17.85 ng/L) and the massage group (171.32 +/- 17.15 ng/L) than that in the control group (220.56 +/- 54.47 ng/L), respectively (P <0.01). There showed no difference in the two aspects between the acupuncture group and the massage group (P >0.05).

CONCLUSIONAcupuncture and massage has a reliable therapeutic effect on primary dysmenorrhea, which is superior to that of Somiton tablet, and its mechanism might be exerted by regulating PGF2alpha level in menstrual fluid.

Acupuncture Therapy ; Decidua ; metabolism ; Dinoprost ; metabolism ; Dysmenorrhea ; therapy ; Female ; Humans ; Massage

OBJECTIVETo investigate relationship between the vascular endothelial growth factor (VEGF) and the pathogenesis of pre-eclampsia in pregnant rats.

METHODSPregnant rats were divided into two groups randomly. Saline solution or L-nitro arginine methyl ester (L-NAME) 125 mg/d was given subcutaneously from day 7 of gestation till establishing pre-eclampsia. Systolic blood pressure, urine protein, platelet count, and weight of pups and placenta were determined. The levels of VEGF in pregnant rats venous serum, placenta and decidual tissue from normal pregnancy and pre-eclampsia rats were detected by ELISA and immunohistochemistry, respectively.

RESULTPregnant rats which were given L-NAME produced physical signs similar to those of pre-eclampsia, such as increase in systolic blood pressure [(145.3 +/-4.6)mmHg] and urine protein [(814.3 +/-57.5)mg/L], and decrease in platelet count [(467.1 +/-76.3) x 10(9)/L] and weight of pups and placenta. Compared with controls, the intensity of VEGF immunostaining in trophoblast or decidual cells were significantly reduced. The serum levels of VEGF were significantly lower in pre-eclampsia group than in normal pregnancy.

CONCLUSIONDecreased serum levels of VEGF and reduced expression of VEGF in placental tissues might in part explain the pathogenesis of pre-eclampsia in pregnant rats.

Animals ; Decidua ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Immunohistochemistry ; NG-Nitroarginine Methyl Ester ; Placenta ; metabolism ; Pre-Eclampsia ; blood ; chemically induced ; metabolism ; Pregnancy ; Random Allocation ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; blood ; metabolism

OBJECTIVETo study the role of platelet-derived growth factor (PDGF) in the pathogenesis of preeclampsia (PRE).

METHODSThirteen normal and 20 PRE late-pregnancy women were enrolled in this study. The serum PDGF-BB levels were measured with enzyme-linked immunosorbent assay, and the expression of PDGF-B mRNA in the decidual blood vessel was determined using in situ hybridization.

RESULTSPDGF-BB levels in PRE group was significantly higher than that in normal pregnant women (83.54 -/+34.52 vs 39.61-/+18.20, P<0.001), and the expression of PDGF-B mRNA in decidual blood vessel was also significantly higher in PRE group (P<0.001), showing a positive correlation between serum PDGF and PDGF-B mRNA expression (r=0.603, P<0.001).

CONCLUSIONPDGF is associated with the pathology of decidual blood vessel. Elevated serum PDGF levels and PDGF-B mRNA expression in the decidual blood vessel may play an important role in the pathogenesis of PRE.

Adult ; Decidua ; blood supply ; Female ; Gene Expression Regulation ; Humans ; Platelet-Derived Growth Factor ; genetics ; metabolism ; Pre-Eclampsia ; blood ; genetics ; metabolism ; pathology ; Pregnancy ; Proto-Oncogene Proteins c-sis ; RNA, Messenger ; genetics ; metabolism