Imprinted genes play a key role in regulating mammalian placental and embryonic development. Here, we generated glutaminyl-peptide cyclotransferase-knockout (Qpct^-/-) mice utilizing the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) platform and identified Qpct as a novel anti-angiogenic factor in regulating mouse placentation. Compared with Qpct^+/+ mice, placentae and embryos (Qpct^-/+ and Qpct^-/-) showed significant overgrowth at embryonic Day 12.5 (E12.5), E15.5, and E18.5. Using single-cell transcriptome analysis of 32 309 cells from Qpct^+/+ and Qpct^-/- mouse placentae, we identified 13 cell clusters via single-nucleus RNA sequencing (snRNA-seq) (8880 Qpct^+/+ and 13 577 Qpct^-/- cells) and 20 cell clusters via single-cell RNA sequencing (scRNA-seq) (6567 Qpct^+/+ and 3285 Qpct^-/- cells). Furthermore, we observed a global up-regulation of pro-angiogenic genes in the Qpct^-/- background. Immunohistochemistry assays revealed a notable increase in the number of blood vessels in the decidual and labyrinthine layers of E15.5 Qpct^-/+ and Qpct^-/- mice. Moreover, the elevation of multiple pairs of ligand-receptor interactions was observed in decidual cells, endothelial cells, and macrophages, promoting angiogenesis and inflammatory response. Our findings indicate that loss of maternal Qpct leads to altered phenotypic characteristics of placentae and embryos and promotes angiogenesis in murine placentae.
Animals ; Female ; Pregnancy ; Mice ; Placentation/genetics* ; Single-Cell Analysis ; Gene Expression Profiling ; Mice, Knockout ; Transcriptome ; Placenta/blood supply* ; Neovascularization, Pathologic/genetics* ; Genomic Imprinting ; Single-Cell Gene Expression Analysis ; Angiogenesis

Alkanesulfonic Acids ; toxicity ; Animals ; Female ; Fluorocarbons ; toxicity ; Humans ; Male ; Mice ; Neovascularization, Physiologic ; drug effects ; Pedigree ; Placenta ; blood supply ; drug effects ; metabolism ; Pregnancy ; Prenatal Exposure Delayed Effects ; genetics ; metabolism ; physiopathology ; RNA, Long Noncoding ; genetics ; metabolism

Preeclampsia (PE) is a pregnancy-specific, multi-system disorder and the leading cause of maternal and perinatal morbidity and mortality in obstetrics worldwide. Excessive vasoconstriction and dysregulated coagulation function are closely associated with PE. Heat shock protein 20 (HSP20) is ubiquitously expressed under normal physiological conditions and has important roles in vascular dilatation and suppression of platelet aggregation. However, the role of HSP20 in the pathogenesis of PE remains unclear. In this study, we collected chorionic plate resistance arteries (CPAs) and serum from 118 healthy pregnant women and 80 women with PE and detected the levels of HSP20 and its phosphorylated form. Both HSP20 and phosphorylated HSP20 were downregulated in CPAs from women with PE. Comparison of the vasodilative ability of CPAs from the two groups showed impaired relaxation responses to acetyl choline in preeclamptic vessels. In addition to the reduced HSP20 in serum from women with PE, the platelet distribution width and mean platelet volume were also decreased, and the activated partial thromboplastin time and thromboplastin time were elevated.With regard to the vital roles of HSP20 in mediating vasorelaxation and coagulation function, the decreased HSP20 might contribute to the pathogenesis of PE.
Adult ; Case-Control Studies ; Chorion ; blood supply ; Female ; HSP20 Heat-Shock Proteins ; metabolism ; Humans ; Phosphorylation ; Placenta ; blood supply ; Platelet Function Tests ; Pre-Eclampsia ; metabolism ; Pregnancy ; Vasoconstriction ; Vasodilation

Placenta is an important organ to maintain fetal growth, metabolism, maternal and fetal physiologic balance. Angiogenesis is a critical factor in placental development involved in fetal blood circulation and vascular changes in the endometrium and placenta. Angiogenesis is closely related to angiogenesis factors such as vascular endothelial growth factor and placenta growth factor. Fetal growth restriction threats the fetal health in gestation and also increases the long-term likeliness of several diseases. In this review, the authors summarize the findings in current studies of the relationship between angiogenesis factors and fetal growth restriction.
Angiogenesis Inducing Agents ; metabolism ; Endometrium ; blood supply ; Female ; Fetal Development ; Fetal Growth Retardation ; Humans ; Neovascularization, Physiologic ; Placenta ; blood supply ; Placenta Growth Factor ; Pregnancy ; Pregnancy Proteins ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo determine the optimal dose of colloid preload, which is both safe and effective, for preventing hypotension in parturients undergoing cesarean section under spinal anesthesia.

METHODSForty-five healthy, termed parturients scheduled for cesarean delivery under spinal anesthesia were randomly assigned to 3 colloid preload groups to receive gelofusine infusion at the rates of 5, 10, or 15 ml·kg(-1)·h(-1) (groups I, II, and III, respectively). Colloid preload was administered 10 min before spinal anesthesia and maintained until the delivery. Blood pressure (BP) and heart rate (HR) of the parturients were monitored during the operation, and Apgar scores at 1 and 5 min after birth were recorded. S100β protein concentration and blood gas values of the umbilical artery were also measured. The vascular adaptation in the placental villous capillary was evaluated stereologically.

RESULTSAt each time point of measurement, BP and HR showed no significant differences among the 3 groups during the operation (P>0.05), but within the same group, BP and HR underwent significant variations during the operation; groups II and III maintained more stable hemodynamics compared to group I. Apgar scores and blood gas analysis, pH value, and S100β protein in the umbilical artery showed no significant differences among the 3 groups (P>0.05). The 3 groups exhibited no significant differences in the length and volume density of the placental villous capillaries (P>0.05).

CONCLUSIONColloid preload with gelofusine administered at the rate of 10 ml·kg(-1)·h(-1) can reduce the incidence and severity of hypotension in cesarean section under spinal anesthesia with the least adverse maternal and fetal effects.

Adult ; Anesthesia, Obstetrical ; Anesthesia, Spinal ; Cesarean Section ; methods ; Colloids ; administration & dosage ; Female ; Fetal Blood ; metabolism ; Humans ; Hypotension ; prevention & control ; Nerve Growth Factors ; blood ; Placenta ; blood supply ; Polygeline ; administration & dosage ; Pregnancy ; S100 Proteins ; blood

Interactions of vascular endothelial growth factor (VEGF) with receptors VEGFR1/Flt1 and VEGFR2/Flk1, and those of angiopoietins (Ang-1, Ang-2) with receptor Tie2 play important roles in placental angiogenesis. This study investigated vascular morphology and expression of these angiogenic factors in rat placenta on the day 15, 18, 21 of gestation (D15, D18 and D21). The rats were randomly assigned into 3 groups: normal group, model group [fetal growth restriction (FGR) model], and Bushen Yiqi Huoxue (BYHR) recipe treatment group (BYHR group, the pregnant rats with FGR were treated with BYHR recipe). Morphological analysis indicated that during initial villous formation, fetal nucleated erythrocytes (FNEs) appeared in maternal blood sinus (MBS). Subsequently, FNEs were surrounded by endothelial cells to form fetal capillary (FC) and then by trophoblast cells to form villi. As pregnancy proceeded, FC density increased progressively with increasing endothelial identification staining (EIS) in normal and BYHR groups. Whereas, villous formation was suppressed, normal increase in FC density was impaired and EIS was weakened in model group. Quantitative PCR analysis showed that VEGF and Flk1 mRNA increased over gestation in all groups, indicating that VEGF might play a pivotal role in FC growth during late gestation. VEGF mRNA was increased on D15, while decreased on D21 in model group as compared with normal group and BYHR group. Immunohistochemically, Ang-2 protein was highly expressed in FNEs, gradually disappeared as villi matured, and decreased over gestation in all groups, indicating that Ang-2 might play a pivotal role in villous formation, which was further supported by decreased Ang-2 mRNA and protein expression in model group on D15. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio increased from D15 to D18 in all groups as placenta matured. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio were decreased on D18 in model group as compared with normal and BYHR groups, indicating delayed maturity of FGR placenta. Alterations in angiogenic factors may result in altered placental vasculature and cause placental insufficiency. BYHR recipe could balance the angiogenic factors to promote the formation and maturation of FGR placental vasculature.
Animals ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Fetal Growth Retardation ; etiology ; physiopathology ; therapy ; Male ; Neovascularization, Physiologic ; drug effects ; physiology ; Placenta ; blood supply ; drug effects ; physiopathology ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Tobacco Smoke Pollution ; adverse effects

OBJECTIVETo explore the optimal approach to the prevention of hypotension during cesarean section for the benefits of both the parturients and the newborns.

METHODSForty singleton full-term pregnant women undergoing elective cesarean delivery were randomly allocated into two equal groups. For prevention of hypotension during spinal anesthesia, ephedrine or pre-anesthetic volume with Voluven was administered. The changes of blood pressure, heart rate, and Apgar scores of the newborns were monitored and recorded, and the umbilical arterial blood gas variables were compared between the two groups. The placental samples were collected and immunohistochemistry for CD34 was performed for stereological study of the placental villous capillaries.

RESULTSThe umbilical arterial PaCO(2), PaO(2) and Apgar scores showed no significant differences between the two groups (P<0.05). The heart rate, incidence of hypotension and the lactic acid value were significantly higher, and the umbilical arterial pH significantly lower in ephedrine group than in the Voluven group (P>0.05). While the length density of the villous capillaries was comparable between the two groups (P>0.05), the volume density of the villous capillaries was significantly decreased in ephedrine group (P<0.05).

CONCLUSIONPre-anesthetic volume expansion with Voluven can maintain stable hemodynamics during spinal anesthesia and also efficiently improve the tissue perfusion, microcirculation and uteroplacental blood flow, thus increasing the oxygen supply to the fetus.

Adult ; Anesthesia, Obstetrical ; adverse effects ; Anesthesia, Spinal ; adverse effects ; Cesarean Section ; Elective Surgical Procedures ; Female ; Humans ; Hydroxyethyl Starch Derivatives ; administration & dosage ; Hypotension ; etiology ; prevention & control ; Placenta ; anatomy & histology ; blood supply ; Placental Circulation ; drug effects ; Plasma Substitutes ; administration & dosage ; Pregnancy

OBJECTIVETo observe the effect of Bushen Yiqi Huoxue Recipe (BYHR, a Chinese medical prescription for reinforcing Shen, replenishing qi and promoting blood circulation) on placental trophoblast apoptosis in fetal growth restricted (FGR) pregnant rat for the sake of explore its mechanism of action in treating FGR.

METHODSFGR animal model was established by passive smoking, 32 pregnant rats were divided into four groups at random: the normal group, the model group, the Chinese medicine (CM) group (model rats treated by BYHR) and the Western medicine (WM) group (model rats treated by arginine). The histological change of placenta was examined with HE stain, the trophoblast apoptosis was detected by TUNEL and RT-PCR, and the expressions of Bcl-2 and Bax in the placenta were detected by immunohistochemical method.

RESULTSBlood stasis and villous ischemia were seen in placenta of FGR model rats. Placental microcirculation was significantly improved in the CM group, but in the WM group only partially improved. The median apoptotic index of syncytial trophoblast cells in the four groups, in normal, model, CM, and WM order, was 45%, 75%, 57% and 70%, as compared with the model group, it was much lower in the normal group and the CM group (P < 0.01), but a similar level was shown in the WM group. No significant difference in mRNA and protein expressions of Bcl-2 and Bax was found among the four groups.

CONCLUSIONBYHR can improve the placental microcirculation in FGR rats to prevent excessive apoptosis through a mechanism other than the classical Bax/Bcl-2 apoptotic pathway, which needs further exploring.

Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Fetal Growth Retardation ; prevention & control ; Microcirculation ; drug effects ; Placenta ; blood supply ; cytology ; Pregnancy ; Rats ; Trophoblasts ; cytology

Abnormal attachment of the placenta (Placenta accreta, increta, and percreta) is an uncommon but potentially lethal cause of maternal mortality from massive postpartum hemorrhage. A 33-yr-old woman, who had been diagnosed with a placenta previa, was referred at 30 weeks gestation. On ultrasound, a complete type of placenta previa and multiple intraplacental lacunae, suggestive of placenta accreta, were noted. For further evaluation of the placenta, pelvis MRI was performed and revealed findings suspicious of a placenta increta. An elective cesarean delivery and subsequent hysterectomy were planned for the patient at 38 weeks gestation. On the day of delivery, endovascular catheters for balloon occlusion were placed within the hypogastric arteries, prior to the cesarean section. In the operating room, immediately after the delivery of the baby, bilateral hypogastric arteries were occluded by inflation of the balloons in the catheters previously placed within. With the placenta retained within the uterus, a total hysterectomy was performed in the usual fashion. The occluding balloons were deflated after closure of the vaginal cuff with hemostasis. The patient had stable vital signs and normal laboratory findings during the recovery period; she was discharged six days after delivery without complications. The final pathology confirmed a placenta increta.
Adult ; Arteries/*surgery ; *Catheterization ; Cesarean Section ; Female ; Gestational Age ; Humans ; Hysterectomy/*methods ; Placenta/*blood supply/ultrasonography ; Placenta Accreta/*surgery/ultrasonography ; Placenta Previa/*surgery/ultrasonography ; Postpartum Hemorrhage/*prevention & control ; Pregnancy ; Treatment Outcome

OBJECTIVE: To investigate the invason of trophoblasts in the placenta bed and the change of spiral arteries and microvessels in pre-eclampsia and normal pregnancy. METHODS: Twenty cases of normal pregnancies, mild pre-eclampsia and severe pre-eclampsia were chosen as Group A, Group B, and Group C. HE staining and immunohistochemistry staining (SP method) were used to observe the depth and the density of trophoblasts invading the placenta bed and the change of spiral arteries and microvessels. RESULTS: The significant difference in the degree of invasion was in the superficial myometrial segment. Group C was the most superficial in the 3 groups (P<0.01). The density of trophoblasts which invaded the placenta bed in the lower half of the basal decidual segment and the myometrial segment showed us Group C was the lowest (P<0.01). There was statistical difference among the 3 groups (P<0.01). The average lumen area of the spiral arteries in the decidual segment and the superficial myometrial segment of the placenta bed was the smallest in Group C among the 3 groups(P<0.01) and there was statistical difference among the 3 groups (P<0.01). The spiral arteries were the thickest in Group C with statistical difference among the 3 groups (P<0.01). The physiological and pathological change of the spiral arteries was mainly in the superficial myometrial segment. The incidence rate of physiological changes in the spiral arteries was the lowest in Group C with statistical difference among the 3 groups (P<0.01). The incidence rate of pathological changes was the highest in Group C (P<0.01) and the normal group was the highest. There was significant difference among the 3 groups(P<0.01). There was positive correlation between the physiological change of the spiral arteries and the invaing degree of the trophoblasts (P<0.05), there was negative correlation between the pathological change of the spiral arteries and the invasion depth as well as the invasion density of the trophoblasts(P<0.05). There was negative correlation between the physiological change and the pathogenetic condition of pre-eclampsia(P<0.05)while there was positive correlation between the pathological change and the pathogenetic condition degree of pre-eclampsia(P<0.05). There was negative correlation between the invasion depth as well as density in uteruso superficial myometrial segment by trophoblast and the pathogenetic condition degree of pre-eclampsia(P<0.05). There was invasion trophoblast in 62.50% lumen wall of spiral arteries in uterus superficial myometrial segment of the placental bed in normal pregnancy while 27.5% was seen in severe pre-eclampsia. Microvascular density in the decidual segment and the superficial myometrial segment of the placenta bed in Group C was the lowest among the 3 groups with statistical difference (P<0.01). CONCLUSION The invasion depth of the trophoblasts in pre-eclampsia was more superficial than normal pregnancy.The changes of the invasion of the trophoblasts and the pathological changes of the spiral arteries in the placenta bed mainly existed in the superficial myometrial segment which was closely related to the severity of the illness. That microvascular density in the placental bed of pre-eclampsia started to decrease from the basal decidual segment shows that the microvessel development in the placenta bed is impaired in pre-eclampsia.
Adult ; Arteries ; pathology ; Capillaries ; pathology ; Female ; Humans ; Placenta ; blood supply ; pathology ; Pre-Eclampsia ; pathology ; Pregnancy ; Trophoblasts ; pathology