Characterization of the folate receptor in human molar placenta

We have characterized a high-affinity folate receptor in human molar placenta tissue. Radioligand binding exhibited characteristics typical of other high-affinity folate binding proteins. Those included, positive cooperativity, a tendency to increased binding affinity with decreasing receptor concentration, a slow ligand dissociation at pH 7.4 becoming rapid at pH 3.5, and inhibition by folate analogues. The folate receptor cross-reacted with antibodies against human milk folate binding protein, e.g. the syncytothrophoblastic layer of molar placenta tissue sections showed strongly positive immunostaining. The gel filtration profile contained two radioligand-bound peaks (25 and 100 kDa), however, with considerable overlap. Only a single band of 70 kDa was seen on SDS-PAGE immunoblotting. The folate receptor in placental tissue may play a crucial role in the transfer of folate from maternal circulation to the fetus.     

Detection of different adenosine triphosphatases in human placental brush border membranes

The microvillous membrane of human placental syncytiotrophoblast cells contains a high ATPase activity. The purpose of this study was to characterize this activity and to investigate the presence of vacuolar type H⁺ ATPase in this membrane. Intact brush border membrane vesicles strongly hydrolyzed ATP, reflecting the presence of ATPase on the external side of the membrane. The ATPase activity was entirely Mg²⁺ dependent and increased with pH. At pH 7.5, Vmax was 31.0 ± 1.7 mol/mg/20 min and Km 0.18 ± 0.03 mm ATP. Hydrolysis of ATP was not influenced by the presence of bicarbonate or alkaline phosphatase inhibitors, but at pH 8 it decreased by half following addition of 100 m dicyclohexylcarbodiimide (DCCD). At pH 7.5, 1 mm N-ethylmaleimide (NEM) depressed this activity by less than 5%. Opening the membrane vesicles with 0.1% desoxycholate (DOC) or Triton-X neither revealed any additional ATPase activity nor altered the low sensitivity to NEM. Treatment of these membranes with 1% cholate decreased the ATPase activity by more than 70% and did not enhance the sensitivity of ATP hydrolysis to NEM. 10^–7 m Bafilomycin, which reduced by 56 ± 9% the ATPase activity in dog kidney brash border membranes treated with 0.1% DOC, had no effect on placental brush border membranes subjected to the same procedure. Finally, neither immunocytochemical staining using monoclonal antibody to the Mr 31000 subunit of V-type H⁺ ATPase, nor electron microscopic examination detected the presence of H⁺ ATPase in placental membranes.In conclusion, the placental brush border membrane is the site of a strong ecto ATPase activity which is partially DCCD sensitive. However, our results did not detect the presence of any vacuolar type H⁺ ATPase activity in these membranes.This study was supported by the MRC grant N MA 9565 and by an extramural grant from Baxter Health Care Corporation. The authors are indepted to Dr. Patrick Vinay for this help in providing Bafilomycin and dog kidney membranes, as well as for his insightful discussion.     

Relationship between glycerol-3-phosphate dehydrogenase,fatty acid synthase and fatty acid binding proteins in developing human placenta

The activities of the enzymes glycerol-3-phosphate dehydrogenase and fatty acid synthase are inhibited by palmitoyl-coenzyme A and oleate. The two isoforms of fatty acid binding proteins (PI 6.9 and PI 5.4) enhance the activities of glycerol-3-phosphate dehydrogenase and fatty acid synthase in the absence of palmitoyl-coenzyme A or oleate and also protect them against palmitoyl-coenzyme A or oleate inhibition. Levels of fatty acid binding proteins, the activities of the enzymes fatty acid synthase and glycerol-3-phosphate dehydrogenase increase with gestation showing a peak at term. However, the activity of fatty acid synthase showed the same trend up to the 30th week of gestation and then declined slightly at term. With the advancement of pregnancy when more lipids are required for the developing placenta, fatty acid binding proteins supply more fatty acids and glycerol-3-phosphate for the synthesis of lipids. Thus a correlation exists between glycerol-3-phosphate dehydrogenase, fatty acid synthase and fatty acid binding proteins in developing human placenta.     

Dose-related action of estradiol on placental prostanoid prediction

Prostanoids play an important role throughout all of pregnancy and during the initiation and progress of labor. The human placenta at term produces large quantities of prostanoids, yet little is known of the factors that regulate their biosynthesis. Herein, we report the effect of estradiol or estradiol and progesterone on the basal release of placental prostanoids from fresh human term placental explants using a perifusion system.The basal release of prostaglandin E₂ (PGE₂, prostaglandin F_2α (PGF_2α), thromboxane (TxB₂) and 6-keto-prostaglandin F_1α (6-keto-PGF_1α) increased about 50% from the fifth to the ninth hour in culture, while the release of 13, 14-dihydro-15-keto-PGF (PGFM) remained constant and hCG release decreased. The dose-related effect of estradiol (20–2,000 ng/ml) in the perifusing medium starting at the fifth hour of perifusiOn (i.e., the zero treatment time) effected no change in the release of TxB₂, PGF_2α, PGFM or hCG. A biphasic action on the release of 6-keto-PGF,. was observed, i.e. it was significantly decreased when incubated with 20 ng/ml of estradiol, but effected an increase after exposure to 200 ng/ml. The concomitant addition of progesterone (2,000 ng/ml) with estradiol (200 ng/ml) significantly inhibited the stimulatory action of estradiol at this dose. The release of PGE₂ was inhibited in a dose-related fashion with increasing dose of estradiol. The addition of progesterone with estradiol (2,000 and 200 ng/ml, respectively) reversed the inhibition of PGE₂ by estradiol alone.These data demonstrate that physiologic levels of estradiol affect 6-keto-PGF_α and PGE₂ release from the human term placenta, but do not significantly alter production of TxB₂, PGFM or hCG under these conditions.     

Transplantation of placenta‐derived mesenchymal stem cells enhances angiogenesis after ischemic limb injury in mice

Mesenchymal stem cell‐based therapy has emerged as a promising approach for the treatment of peripheral arterial disease. The purpose of this study was to examine the potential effects of human placenta‐derived mesenchymal stem cells (PMSCs) on mouse hindlimb ischemia. PMSCs were isolated from human placenta tissue and characterized by flow cytometry. An in vivo surgical ligation‐induced murine limb ischemia model was generated with fluorescent dye (CM‐DiI) labelled PMSCs delivered via intramuscular injection. Our data show that PMSCs treatment significantly enhanced microvessel density, improved blood perfusion and diminished pathologies in ischemic mouse hindlimbs as compared to those in the control group. Further immunostaining studies suggested that injected PMSCs can incorporate into the vasculature and differentiate into endothelial and smooth muscle cells to enhance angiogenesis in ischemic hind limbs. This may in part explain the beneficial effects of PMSCs treatment. Taken together, we found that PMSCs treatment might be an effective treatment modality for treatment of ischemia‐induced injury to mouse hind limbs by enhancement of angiogenesis.     

Telocytes in female reproductive system (human and animal)

Telocytes (TCs) are a newly discovered type of cell with numerous functions. They have been found in a large variety of organs: heart (endo‐, myo‐, epi‐ and pericardium, myocardial sleeves, heart valves); digestive tract and annex glands (oesophagus, stomach, duodenum, jejunum, liver, gallbladder, salivary gland, exocrine pancreas); respiratory system (trachea and lungs); urinary system (kidney, renal pelvis, ureters, bladder, urethra); female reproductive system (uterus, Fallopian tube, placenta, mammary gland); vasculature (blood vessels, thoracic duct); serous membranes (mesentery and pleura); and other organs (skeletal muscle, meninges and choroid plexus, neuromuscular spindles, fascia lata, skin, eye, prostate, bone marrow). Likewise, TCs are widely distributed in vertebrates (fish, reptiles, birds, mammals, including human). This review summarizes particular features of TCs in the female reproductive system, emphasizing their involvement in physiological and pathophysiological processes.     

子宫动脉栓塞术在胎盘前置状态中孕引产的疗效观察

目的:探讨子宫动脉栓塞术在胎盘前置状态中期妊娠引产中的应用。方法:选择2014年1月~2016年1月我院收治的孕14~24周胎盘前置状态需要终止妊娠患者32例,随机分为两组,其中15例先行子宫动脉栓塞术,3小时后给予利凡诺100 mg羊膜腔内注射引产术,联合口服米非司酮150 mg(A组),17例行常规剖宫取胎术(B组),比较两组的出血量及住院时间。结果:A组产后平均出血量为284.53±33.74 m L,平均住院时间为3.8±0.7天;B组平均产后出血量为546.07±40.69 m L,平均住院时间为6.4±0.3天,差异均有统计学意义(P0.05)。结论:子宫动脉栓塞术可用于胎盘前置状态的中孕引产,而且具有出血少、创伤小、可保护女性生殖生理功能的优点,值得临床推广。     

表达K1-5 蛋白的人胎盘间充质干细胞抑制大鼠主动脉环血管生成的实验研究

目的:观察表达外源性Kringle1-5(K1-5)蛋白的人胎盘组织的间充质干细胞(HPMSCs)在体外对大鼠主动脉环血管生成的影响。方法:用胶原酶和贴壁法从人胎盘组织中提取间充质干细胞。选取感染复数MOI:50,将重组腺病毒载体rAd-K1-5感染HPMSCs至48 h;应用荧光显微镜观察转染效率。取8周雌性SD大鼠的腹主动脉,建立主动脉环血管生成体外模型,6天后观察基质胶内微血管形成的情况。结果:从人胎盘组织提取的成纤维样细胞具有贴壁生长特性,可向成骨细胞、脂肪细胞分化,证明从胎盘提取的细胞是间充质干细胞。采用rAd-K1-5腺病毒载体感染的HPMSCs可分泌Kringle1-5蛋白,表达K1-5蛋白的转基因HPMSCs处理的基质胶栓中管样结构的形成明显减少。结论:表达外源性Kringle1-5蛋白的HPMSCs能够体外抑制大鼠主动脉环新生血管生成。