Adult ; Diabetes, Gestational ; metabolism ; Female ; Humans ; Matrix Metalloproteinases ; metabolism ; Placenta ; enzymology ; Pregnancy

C-Terminal carboxyl methylation of a human placental 23 kDa protein catalyzed by membrane-associated methyltransferase has been investigated. The 23 kDa protein substrate methylated was partially purified by DEAE-Sephacel, hydroxyapatite and Sephadex G-100 gel filtration chromatographies. The substrate protein was eluted on Sephadex G-100 gel filtration chromatography as a protein of about 29 kDa. In the absence of Mg2+, the methylation was stimulated by guanine nucleotides (GTP, GDP and GTPgammaS), but in the presence of Mg2+, only GTPgammaS stimulated the methylation which was similar to the effect on the G25K/rhoGDI complex. AFC, an inhibitor of C-terminal carboxyl methylation, inhibited the methylation of human placental 23 kDa protein. These results suggests that the substrate is a small G protein different from the G25K and is methylated on C-terminal isoprenylated cysteine residue. This was also confirmed by vapor phase analysis. The methylated substrate protein was redistributed to membrane after in vitro methylation, suggesting that the methylation of this protein is important for the redistribution of the 23 kDa small G protein for its putative role in intracellular signaling.
Cysteine/metabolism* ; Female ; GTP-Binding Proteins/metabolism* ; Guanine Nucleotides/pharmacology ; Human ; Methylation ; Placenta/metabolism* ; Placenta/enzymology ; Pregnancy ; Pregnancy Proteins/metabolism* ; Protein Methyltransferases/metabolism*

OBJECTIVE: To investigate the effect of angiotensin converting enzyme (ACE) in the pathogenesis of preeclampsia. METHODS: A cross-sectional study was conducted with 42 pregnant women in the following categories: 30 cases of preeclampsia (mild preeclampsia, n=15; severe preeclampsia, n=15), and normal pregnancy (control group,n=12). The expression and localization of ACE mRNA in the placenta of the 3 groups were respectively examined by in situ hybridization. Ultraviolet radiation colorimetry was used to detect the activity of ACE in the placenta tissue homogenate and the mothers' serum in the 3 groups. RESULTS: The expression of ACE mRNA was found in the endothelial cells of villus and trophoblasts in the placenta. The positive index of ACE mRNA in the placenta of preeclampsia(3.12+/-0.94) was higher than that in the normal pregnancies(1.65+/-0.67) (P<0.05), and there was significant difference between severe preeclampsia and mild preeclampsia (P<0.05). The levels of ACE activity in the placenta tissue homogenate and the maternal serum of preeclampsia were higher than those in the normal pregnancies (P<0.05), and there was significant difference between severe preeclampsia and mild preeclampsia (P<0.05). The placenta tissue homogenate ACE activity was correlated with ACE activity of the maternal serum (r=0.781,P<0.05). CONCLUSION The expression and activity of local ACE in the placenta tissue may play an important role in preeclampsia and contribute to the development of preeclampsia.
Adult ; Cross-Sectional Studies ; Female ; Humans ; In Situ Hybridization ; Peptidyl-Dipeptidase A ; blood ; genetics ; metabolism ; Placenta ; enzymology ; Pre-Eclampsia ; enzymology ; Pregnancy ; RNA, Messenger ; biosynthesis ; genetics

The effects of ethyl alcohol and pig serum administration on the development of preneoplastic hepatic enzyme-altered foci were examined in an in vivo mid-term assay system. Rats were initially given a single dose (200 mg/Kg) intraperitoneal injection of diethylnitrosamine (DEN). Two weeks later, treatment was started with 10% ethanol + 10% sucrose solution, 10% sucrose solution, or tap water as drinking water for 6 weeks with or without intraperitoneal injection of porcine serum twice a week. All rats were subjected to a two-thirds partial hepatectomy at week 3. The modification potentials were evaluated by comparing the number and area per cm2 of glutathione S-transferase placental form-positive (GST-P+) foci in the liver of each group. As a result, ethanol significantly enhanced the development of GST-P+ foci. Unfortunately, the porcine serum injection produced no hepatic fibrosis and no significant alteration in GST-P+ foci.
Animals ; Diethylnitrosamine/*toxicity ; Ethanol/*pharmacology ; Glutathione Transferase/*metabolism ; Immune Sera/pharmacology ; Liver Cirrhosis, Alcoholic/enzymology ; Male ; Placenta/drug effects/*enzymology ; Precancerous Conditions/*chemically induced/enzymology ; Rats ; Rats, Inbred F344 ; Survival Rate ; Swine

The enzyme complex 3b-hydroxysteroid dehydrogenase/delta(5)-delta(4)-isomerase (3beta-HSD) is involved in the biosynthesis of all classes of active steroids. The expression of 3beta-HSD in human uterine endometrium during the menstrual cycle and decidua was examined in an effort to understand its role during ova implantation. 3beta-HSD was weakly expressed in the glandular epithelium of the proliferative phase and moderately expressed in the glandular epithelium of secretory phase of the endometrium. In the decidua of the ectopic pregnancy, 3beta-HSD was strongly expressed. The human uterine endometrial 3beta-HSD was identified as being the same type as the placental 3beta-HSD by RT-PCR and sequence analysis. In addition to the expression of 3beta-HSD, P450scc was expressed in the decidua of the ectopic pregnancy. These results suggest that pregnenolone might be synthesized from cholesterol by P450scc de novo and then, it is converted to progesterone by 3beta-HSD in the uterine endometrium. The data implies that the endometrial 3beta-HSD can use not only the out-coming pregnenolone from the adrenal gland but also the self- made pregnenolone to produce progesterone. The de novo synthesis of progesterone in the endometrium might be a crucial factor for implantation and maintenance of pregnancy.
Cholesterol/chemistry ; Cholesterol Side-Chain Cleavage Enzyme/*biosynthesis/genetics ; Decidua/enzymology ; Endometrium/*enzymology ; Female ; Gene Expression/physiology ; Human ; Menstrual Cycle/physiology ; Multienzyme Complexes/*biosynthesis/genetics ; Placenta/enzymology ; Pregnancy ; Pregnenolone/biosynthesis ; Progesterone/biosynthesis ; Progesterone Reductase/*biosynthesis/genetics ; Steroid Isomerases/*biosynthesis/genetics

OBJECTIVETo test the possibility that certain efficient substrates for lipoxygenase (LOX) produce shuttle oxidants that stimulate the generation of reactive oxygen species from other chemicals.

METHODSMetabolic interactions were conducted in vitro between chlorpromazine (BZ) or other phenothiazines and benzidine or other xenobiotics mediated by soybean lipoxygenase (SLO) or purified human term placental lipoxygenase (HTPLO) in the presence of hydrogen peroxide, and the rates of xenobiotics oxidation were measured by spectroscopic analysis.

RESULTSChlorpromazine cation radical (CPZ(*+)) generated by lipoxygenase triggered a rapid oxidation of benzidine to benzidine diimine cation. Under the experimental conditions used, the metabolic interaction resulted in a 42-fold greater stimulation than BZ alone in the rate of BZ oxidation. The magnitude of stimulation of benzidine oxidation exhibited a dependence on the pH of the reaction medium, amount of the enzyme, and concentration of chlorpromazine, BZ, and hydrogen peroxide. A number of other phenothiazines were also found to stimulate BZ oxidation, albeit to a lesser degree. Chlorpromazine cation radical stimulated the oxidation of all six other xenobiotics tested. The highest stimulation (94-fold) was noted in the oxidation of tetramethyl phenylenediamine to Wursters blue radical, while the lowest stimulatory response (2-fold) was observed in guaiacol. Preliminary data showed that purified HTPLO also displayed a similar stimulatory response to benzidine oxidation in the presence of chlorpromazine.

CONCLUSIONBoth soybean lipoxygenase and purified human term placental lipoxygenase can mediate stimulatory response to the oxidation of benzidine and other xenobiotics in the presence of phenothiazines.

Benzidines ; metabolism ; Chlorpromazine ; chemistry ; Drug Interactions ; Female ; Humans ; Lipoxygenase ; isolation & purification ; pharmacology ; Oxidation-Reduction ; drug effects ; Phenothiazines ; pharmacology ; Placenta ; enzymology ; Pregnancy ; Xenobiotics ; metabolism

To evaluate the induction of preneoplastic hepatic foci in relation to natural killer cell (NK) activity, we sequentially analyzed glutathione S-transferase placental form positive (GST-P+) hepatocytes and NK activity during diethylnitrosamine (DEN) and phenobarbital (PB)-induced hepatocarcinogenesis in Sprague-Dawley rats. Previous studies have shown that NK activity can modulate the carcinogenic process induced by chemical carcinogens. Newborn females were initially given a single intraperitoneal injection of 15 mg DEN/kg and three weeks later, they were treated with 500 ppm phenobarbital (PB). From week 3, PB was administered in drinking water for 9 weeks. Interim and terminal sacrifices were performed at weeks 12, 15 and 30. GST-P+ hepatocytes increased with age in DEN-treated rats, especially in the population of more than two GST-P+ hepatocytes. The NK activity of DEN-treated rats did not significantly differ from that of control rats until week 12, but it progressively decreased from week 15 to 30. These results indicate that changes of NK activity inversely correlated with the induction of preneoplastic hepatic foci. This strong correlation of decreased NK activity with enhanced induction of GST-P+ foci suggests that NK activity is important in the early progression of hepatocarcinogenesis in rats.
Animal ; Body Weight ; Carcinogens/pharmacology* ; Diethylnitrosamine/pharmacology* ; Female ; Glutathione Transferase/metabolism* ; Killer Cells, Natural/immunology* ; Liver/enzymology* ; Liver/cytology ; Liver Neoplasms/physiopathology* ; Organ Weight ; Placenta ; Rats ; Rats, Sprague-Dawley

In order to provide a new method for the identification of Placenta hominis, the COI barcode has been employed to identify the P. hominis medicinal materials and its adulterants. Genomic DNA was extracted from the experimental samples. The COI sequences were amplified and sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. NJ tree was constructed by MEGA6.0 software. COI sequences can be successfully obtained from all experimental samples. The intra-specific variation and inter-specific divergence were calculated. The average intra-specific K2P distance of P. hominis was 0.001 and the maximum intra-specific distance was 0.008. The cluster dendrogram constructed can be seen that the same genus is together, and distinguished from its adulterants. It is concluded that P. hominis and its adulterants can be correctly identified by DNA barcoding method.
Animals ; Cattle ; DNA Barcoding, Taxonomic ; methods ; Drug Contamination ; prevention & control ; Electron Transport Complex IV ; genetics ; Female ; Humans ; Medicine, Chinese Traditional ; Molecular Sequence Data ; Phylogeny ; Placenta ; chemistry ; enzymology ; Pregnancy ; Quality Control ; Sheep ; Swine

The effect of astaxanthin on N(Ω)-nitro-L-arginine methyl ester (L-NAME) induced preeclampsia disease rats was investigated. Thirty pregnant Sprague-Dawley rats were randomly divided into three groups (n = 10): blank group, L-NAME group and astaxanthin group. From day 5 to 20, astaxanthin group rats were treated with astaxanthin (25 mg x kg(-1) x d(-1) x bw(-1)) from pregnancy (day 5). To establish the preeclamptic rat model, L-NAME group and astaxanthin group rats were injected with L-NAME (125 mg x kg(-1) x d(-1) x bw(-1)) from days 10-20 of pregnancy. The blood pressure and urine protein were recorded. Serum of each group was collected and malondialdehyde (MDA), superoxide dismutase (SOD) and nitric oxide synthase (NOS) activities were analyzed. Pathological changes were observed with HE stain. The expression of NF-κB (nuclear factor kappa B), ROCK II (Rho-associated protein kinase II), HO-1 (heme oxygenase-1) and Caspase 3 were analyzed with immunohistochemistry. L-NAME induced typical preeclampsia symptoms, such as the increased blood pressure, urinary protein, the content of MDA, etc. Astaxanthin significantly reduced the blood pressure (P < 0.01), the content of MDA (P < 0.05), and increased the activity of SOD (P < 0.05) of preeclampsia rats. The urinary protein, NO, and NOS were also decreased. HE stain revealed that after treated with astaxanthin, the thickness of basilal membrane was improved and the content of trophoblast cells and spiral arteries was reduced. Immunohistochemistry results revealed that the expressions of NF-κB, ROCK II and Caspase 3 in placenta tissue were effectively decreased, and HO-1 was increased. Results indicated that astaxanthin can improve the preeclampsia symptoms by effectively reducing the oxidative stress and inflammatory damages of preeclampsia. It revealed that astaxanthin may be benefit for prevention and treatment of preeclampsia disease.
Animals ; Blood Pressure ; Caspase 3 ; metabolism ; Disease Models, Animal ; Female ; Heme Oxygenase (Decyclizing) ; metabolism ; Malondialdehyde ; metabolism ; NF-kappa B ; metabolism ; NG-Nitroarginine Methyl Ester ; Nitric Oxide Synthase ; metabolism ; Oxidative Stress ; Placenta ; enzymology ; Pre-Eclampsia ; drug therapy ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Xanthophylls ; therapeutic use ; rho-Associated Kinases ; metabolism

Axl encodes the tyrosine-protein kinase receptor, participating in the proliferation and migration of many cells. This study examined the role of Axl in functions of endothelial progenitor cells (EPCs). Axl was detected by RT-PCR and Western blotting in both placentas and EPCs from normal pregnancy and preeclampsia patients. The Axl inhibitor, BMS777-607, was used to inhibit the Axl signalling pathway in EPCs. Cell proliferation, differentiation, migration and adhesion were measured by CCK-8 assay, cell differentiation assay, Transwell assay, and cell adhesion assay, respectively. Results showed the expression levels of Axl mRNA and protein were significantly higher in both placentas and EPCs from preeclampsia patients than from normal pregnancy (P<0.05). After treatment with BMS777-607, proliferation, differentiation, migration and adhesion capability of EPCs were all significantly decreased. Our study suggests Axl may play a role in the function of EPCs, thereby involving in the pathogenesis of preeclampsia.
Adult ; Aminopyridines ; pharmacology ; Blood Pressure ; Case-Control Studies ; Cell Adhesion ; drug effects ; Cell Differentiation ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Female ; Fetal Blood ; cytology ; enzymology ; Gene Expression Regulation ; Gestational Age ; Human Umbilical Vein Endothelial Cells ; drug effects ; enzymology ; pathology ; Humans ; Placenta ; metabolism ; physiopathology ; Pre-Eclampsia ; blood ; genetics ; physiopathology ; Pregnancy ; Primary Cell Culture ; Protein Kinase Inhibitors ; pharmacology ; Proto-Oncogene Proteins ; antagonists & inhibitors ; genetics ; metabolism ; Pyridones ; pharmacology ; RNA, Messenger ; antagonists & inhibitors ; genetics ; metabolism ; Receptor Protein-Tyrosine Kinases ; antagonists & inhibitors ; genetics ; metabolism ; Stem Cells ; drug effects ; enzymology ; pathology