The effect of prostacyclin on the human umbilical artery

Prostacyclin was tested on human umbilical artery obtained after spontaneous delivery or by Cesarean section. Isometric and isotonic responses were measured on spiral preparations in Krebs-bicarbonate buffer at 37°C equilibrated with 95% O₂ and 5% CO₂. Spiral artery strips, whether superfused or mounted in organ baths isometrically or isotonically, responded in a dose-dependent manner to both prostacyclin and serotonin; the PGI₂ response was biphasic in that low doses (2.5 × 10^-8 M - 1.0 × 10^-6 M) elicited a dose-dependent relaxation which changed with higher concentrations (1.0 × 10^-6 M - 2.53 × 10⁵ M) to a contractile response. The maximum tension exerte was 50% less than that elicited by serotonin. The data indicate that the human umbilical artery is responsive to prostacyclin and may be involved in the regulation of fetal placenta blood flow.     

The effect of prostacyclin on the human umbilical artery.

Prostacyclin was tested on human umbilical artery obtained after spontaneous delivery or by Cesarean section. Isometric and isotonic responses were measured on spiral preparations in Krebs-bicarbonate buffer at 37 degrees C equilibrated with 95% O2 and 5% CO2. Spiral artery strips, whether superfused or mounted in organ baths isometrically or isotonically, responded in a dose-dependent manner to both prostacyclin and serotonin; the PGI2 response was biphasic in that low doses (2.5 x 10(-8) M -1.0 x 10(-6) M) elicited a dose-dependent relaxation which changed with higher concentrations (1.0 x 10(-6) M -2.53 X 10(-5) M) to a contractile response. The maximum tension exerted was 50% less than that elicited by serotonin. The data indicate that the human umbilical artery is responsive to prostacyclin and may be involved in the regulation of fetal placenta blood flow.     

Protective effect of melatonin on myocardial infarction

The dose- and time dependence of melatonin and the effective window of melatonin administration were determined in a mouse model of myocardial infarction. When mouse hearts were subjected to 60 min of occlusion of the left anterior descending artery (LAD) followed by 4 h of reperfusion, melatonin pretreatment for 30 min significantly reduced the infarct size/risk area. The most effective dose was found to be 150 microg/kg intraperitoneally, and the effective period of protection lasted up to 2 h after melatonin administration. Melatonin administration 45 min after LAD ligation or right before reperfusion was as effective as administration 30 min before ligation; however, melatonin administered after the release of occlusion was not protective. Melatonin's effect was still present in mice deficient for the Mel1a melatonin receptor. 8-Methoxy-2-propionamidotetralin, a melatonin receptor agonist with no antioxidant activity, offered no protection, suggesting a lack of involvement of melatonin receptors. Finally, the effects of melatonin were similar in rats and mice. Our results demonstrate that melatonin is an effective cardioprotective agent when administered either before or during coronary occlusion at a very low dose.     

Melatonin counteracts potentiation by homocysteine of KCL-induced vasoconstriction in human umbilical artery: relation to calcium influx

Homocysteinemia is a major and independent risk factor for vascular disease. Oxidative stress is a possible mechanism for homocysteine (HCY)-induced vascular disease. Herein, we evaluated the antioxidant property of melatonin (MLT) in relation to the vasoconstrictive effect of HCY on the human umbilical artery. Helical umbilical arterial strips without endothelium were obtained at elective Cesarean delivery near term. Changes in potassium chloride (KCl)-induced vasoconstriction were measured. Arterial strips were treated with HCY (10 or 100 microM) plus FeSO(4) (10 microM) alone or pretreated with a hydroxyl radical ((*)OH) scavenger, mannitol (20 mM), or MLT (1 or 10 microM). The effect of HCY on the response of arterial strips to external calcium (Ca(2+)) in the presence of KCl (20 mM) was determined. HCY plus FeSO(4) potentiated KCl-induced vasoconstriction in a concentration-dependent manner; pretreatment with mannitol significantly reduced this vasospastic effect. HCY (100 microM) significantly augmented the contractile response to external Ca(2+). MLT (10 microM) significantly suppressed the contractile response to external Ca(2+). These results suggest that HCY potentiates KCl-induced umbilical artery vasoconstriction, in part by increasing Ca(2+) influx in vascular smooth muscle cells via activation of Ca(2+) channels. MLT significantly suppressed the vasoconstrictive effect of HCY, probably by scavenging (*)OH arising from HCY autooxidation.     

Protective effect of a calcium antagonist against renal vasoconstriction induced by endothelin in the normotensive rat

We studied the effect of nifedipine, a dihydropyridine calcium antagonist, on the hemodynamic changes induced by endothelin, in awake normotensive rats. Endothelin (0.07-1.40 nmol/kg, e.v.) caused an initial hypotensive effect, followed by long lasting hypertension. Renal blood flow was reduced immediately and still remained below basal levels, at 30 minutes after endothelin injection. Nifedipine (1 mg/kg, i.p.) significantly prevented the effect of endothelin on mean blood pressure and induced a right-ward shift in the dose response curve of renal hemodynamic changes induced by endothelin. We conclude that treatment with calcium antagonist could be very useful in all those conditions in which systemic and regional vasocostriction is provoked by endothelin.     

Protective effect of melatonin on zymosan-induced cellular damage

We investigated whether in vivo melatonin treatment inhibits cellular injury induced by peroxynitrite production and PARS activation in macrophages collected from rats subjected to zymosan-induced shock. Macrophages harvested from the peritoneal cavity exhibited a significant production of peroxynitrite, as measured by the oxidation of the fluorescent dye dihydrorhodamine 123. Furthermore, zymosan-induced shock suppressed macrophage mitochondrial respiration, DNA strand breakage, activation of the nuclear enzyme poly(ADP-ribose)synthetase (PARS) and reduction of cellular levels of NAD+. In vivo treatment with melatonin (25 and 50 mg/kg, i.p., 1 h after zymosan injection) significantly and dose-dependently reduced peroxynitrite formation and prevented the appearance of DNA damage, decrease in mitochondrial respiration, loss of cellular levels of NAD+ and PARS activation. Our study supports the view that the antioxidant and anti-inflammatoy effect of melatonin is also correlated with the inhibition of peroxynitrite production and PARS activation. In conclusion, melatonin may be a novel pharmacological approach to prevent cell injury in inflammation.     

Protective effects of melatonin against carbon tetrachloride hepatotoxicity in rats

Melatonin is an indolamine, mainly secreted by the pineal gland into the blood of mammalian species. The potential for protective effects of melatonin on carbon tetrachloride (CCl(4))-induced acute liver injury in rats was investigated in this work. CCl(4) exerts its toxic effects by generation of free radicals; it was intragastrically administered to male Wistar rats (4 g kg(-1) body weight) at 20 h before the animals were decapitated. Melatonin (15 mg kg(-1) body weight) was administered intraperitoneally three times: 30 min before and at 2 and 4 h after CCl(4) injection. Rats injected with CCl(4) alone showed significant lipid and hydropic dystrophy of the liver, massive necrosis of hepatocytes, marked increases in free and conjugated bilirubin levels, elevation of hepatic enzymes (alanine aminotransferase and aspartate aminotransferase) in plasma, as well as NO accumulation in liver and in blood. Melatonin administered at a pharmacological dose diminished the toxic effects of CCl(4). Thus it decreased both the structural and functional injury of hepatocytes and clearly exerted hepatoprotective effects. Melatonin administration also reduced CCl(4)-induced NO generation. These findings suggest that the effect of melatonin on CCl(4)-induced acute liver injury depends on the antioxidant action of melatonin.     

Protective effect of melatonin against propoxur-induced oxidative stress and suppression of humoral immune response in rats

Effect of melatonin in attenuation of propoxur induced oxidative stress and suppression of humoral immune response was studied in rats. Oral administration of propoxur (10 mg/kg) increased lipid peroxidation in serum after 28 days treatment. Superoxide dismutase, catalase and glutathione were also altered following propoxur exposure. In addition propoxur exposure markedly suppressed humoral immune response as assessed by antibody titre and plaque forming cell assay. Simultaneous treatment with melatonin (5 mg/kg, ip) markedly attenuated the effect of propoxur on (a) lipid peroxidation, (b) oxidative stress parameters and (c) immunotoxicity. Results have been discussed in the light of possible immunopotentiating and antioxidant effects of melatonin to understand the influence of oxidative stress on propoxur induced immunomodulation.     

Protective effects of peoniflorin against hydrogen peroxide-induced oxidative stress in human umbilical vein endothelial cells

Peoniflorin (PF), extracted from the root of Paeonia lactiflora Pall., has been reported to have anti-inflammation and antioxidant effects in several animal models. Herein, we investigated the protective effects of PF against hydrogen peroxide (H(2)O(2))-induced oxidative damage in human umbilical vein endothelial cells (HUVECs). HUVECs were treated by H(2)O(2) (240?μmol/L) with or without PF. PF significantly increased the percent cell viability of HUVECs injured by H(2)O(2) using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. By flow cytometric analysis, PF markedly attenuated H(2)O(2)-induced apoptosis and intracellular reactive oxygen species production. In addition, PF also displayed a dose-dependent reduction of lactate dehydrogenase leakage, malondialdehyde formation, and caspase-3 proteolytic activities in H(2)O(2)-treated cells, which was accompanied with a restoration of the activities of endogenous antioxidants, including total superoxide dismutase and glutathione peroxidase. Finally, Western blot data revealed that H(2)O(2) upregulated phosphorylation of extracellular signal-regulated kinase 1/2 in HUVECs, which was almost completely reversed by PF. Taken together, our data provide the first evidence that PF has a protective ability against oxidative damage in HUVECs. PF may be a candidate medicine for the treatment of vascular diseases associated with oxidative stress.     

Prostanoid synthesis by human umbilical artery

A discrepancy between published values of PGI2 production by human umbilical artery in vitro measured by platelet bioassay, compared with values of 6-oxo-PGF1 alpha by radioimmunoassay, raised the possibility that another anti-aggregatory prostanoid was produced by this tissue. To test this hypothesis, umbilical artery rings were incubated in buffer and PGI2 determined by platelet bioassay and by a more specific radioimmunoassay based on comparison of 6-oxo-PGF1 alpha in hydrolysed and non-hydrolysed samples. 6-oxo-PGF1 alpha, PGF2 alpha and TXB2 were also measured by gas chromatography negative ion chemical ionisation mass spectrometry. PGI2 concentrations by radioimmunoassay and bioassay were significantly correlated (r = 0.92, p less than 0.01). There was no difference between them, disproving the presence of an additional antiaggregatory substance. PGI2 production determined by bioassay (mean 1.21 ng/mg wet weight/h, range 0.59-1.53 ng/mg/h) differed from previously reported values (range 70-325 ng/mg/h). 6-oxo-PGF1 alpha concentrations were confirmed by gas chromatography negative ion chemical ionisation mass spectrometry. Previous determinations of PGI2 production by this tissue overestimated it by approximately 100 times.     

Prostanoid synthesis by human umbilical artery

A discrepancy between published values of PGI₂ production by human umbilical artery measured by platelet bioassay, compared with values of 6-oxo-PGF_1α by radioimmunoassay, raised the possibility that another anti-aggregatory prostanoid was produced by this tissue. To test this hypothesis, umbilical artery rings were incubated in buffer and PGI₂ determined by platelet bioassay and by a more specific radioimmunoassay based on comparison of 6-oxo-PGF_1α in hydrolysed and non-hydrolysed samples. 6-oxo-PGF_1a, PGF_2α and TXB₂ were also measured by gas chromatography negative ion chemical ionisation mass spectrometry. PGI₂ concentrations by radioimmunoassay and bioassay were significantly correlated (r = 0.92, p < 0.01). There was no difference between them, disproving the presence of an additional antiaggregatory substance. PGI₂ production determined by bioassay (mean 1.21 ng/mg wet weight/h, range 0.59–1.53 ng/mg/h) differed from previously reported values (range 70–325 ng/mg/h). 6-oxo-PGF_1α concentrations were confirmed by gas chromatography negative ion chemical ionisation mass spectrometry. Previous determinations of PGI₂ production by this tissue overestimated it by approximately 100 times.     

Protective effect of ginkgo biloba against gossypol-induced apoptosis in human lymphocytes

Ginkgo biloba (EGb 761) is a well-defined plant extract that directly scavenges hydroxyl radicals. It is a potent antioxidant that inhibits apoptosis in cultured cells and is effective in treating mild-to-moderate dementia in Alzheimer patients. Apoptosis is an active process of cell destruction and it plays an important role in pathological processes. The aim of this study is to investigate the anti-apoptotic effect of EGb 761 in gossypol-treated human lymphocytes. Pretreatment of lymphocytes with 10 microg/ml EGb 761 for 30 min or 1h decreased the percentage of apoptosis to 17.5% and 20%, respectively. EGb 761 treatment (25-150 microg/ml) decreased the level of apoptosis to a plateau between 8 and 10% of the control values. We conclude that EGb 761 reduces gossypol-induced apoptosis in human lymphocytes.     

Element content of human umbilical artery and vein in umbilical cord

To elucidate the element content of newborn blood vessels, umbilical arteries and veins in human umbilical cords, which had the advantage of easy sampling, were examined by ICP-AES. Umbilical cords were removed after birth. Mothers’ ages ranged from 26 to 35 yr. It was found that the content of sulfur was the highest in both umbilical arteries and veins, being higher than the content of calcium and phosphorus. With respect of the content of sulfur, calcium, and magnesium, there were significant differences between the arteries and veins.     

金属硫蛋白拮抗同型半胱氨酸对血管内皮细胞的脂质过氧化作用

目的:观察金属硫蛋白(metallothionein,MT)对同型半胱氨酸(homocysteine Hcy)损伤血管内皮细胞的拮抗作用及机制.方法:培养液中分别加入Hcy及Hcy MT孵育血管内皮细胞,自动生化分析仪测培养液乳酸脱氢酶(LDH)活性,硫代巴比妥酸法测细胞丙二醛(MDA)含量.结果:同型半胱氨酸增加内皮细胞LDH及蛋白漏出,并使细胞MDA含量明显升高.MT呈浓度依赖性地抑制Hcy所致的血管内皮细胞损伤及MDA增高.结论:MT拮抗Hcy对血管内皮细胞的脂质过氧化损伤.     

Protective role of mast cells in homocysteine-induced cardiac remodeling

Recent reports including those from our laboratories indicate that hyperhomocysteinemia (Hhe) is an independent risk factor for cardiac dysfunction and clinical heart failure. Mast cell accumulation is a prominent feature in our model of Hhe-induced cardiac dysfunction. Because mast cell-derived mediators can potentially attenuate cardiac remodeling, we investigated the possible protective role of mast cells in Hhe-induced cardiac remodeling using a mast cell-deficient rat model that in our recent report did not demonstrate any adverse cardiac function at younger age (6 mo) than mast cell-competent control animals. Mast cell-deficient (Ws/Ws) rats and mast cell-competent (+/+) littermate control animals (3 mo of age) were treated with a Hhe-inducing diet for 10 wk. Cardiac remodeling was assessed structurally utilizing histomorphometric methods and functionally using an isolated Langendorff-perfused heart preparation. The Hhe-inducing diet caused similar elevations of homocysteine levels in the two groups. Compared with Hhe +/+ rats, the Hhe Ws/Ws rats demonstrated strikingly exacerbated adverse cardiac remodeling and myocardial fibrosis. Cardiac function measurement showed worsened diastolic function in Hhe Ws/Ws rats compared with Hhe +/+ rats. The absence of mast cells strikingly exacerbates Hhe-induced adverse cardiac remodeling and diastolic dysfunction. These findings indicate a potential dual rather than sole deleterious role for mast cells in cardiac injury.     

Protective effect of melatonin against transient global cerebral ischemia-induced neuronal cell damage via inhibition of matrix metalloproteinase-9

Aims

Melatonin possesses various pharmacological effects including neuroprotective effects against brain ischemia. Post-ischemic increases in matrix metalloproteinase-9 (MMP-9) expression and activity mainly contribute to neuronal damage by degradation of the extracellular matrix. This study was designed to examine whether melatonin has a neuroprotective effect and an influence on MMP-9 in transient global brain ischemia.

Main methods

Mice were subjected to 20 min of global brain ischemia and sacrificed 72 h later. Melatonin (30 mg/kg) was administered 30 min before and 2 h after ischemia as well as once daily until sacrifice.

Key findings

Hippocampal pyramidal cell damage after ischemia was significantly decreased by melatonin. As observed by zymography, melatonin inhibited the increase of MMP-9 activity after ischemia. In the brain sections, the increased gelatinase activity was mainly observed in the hippocampus after ischemia and melatonin also reduced gelatinase activity. The laminin and NeuN expression levels were reduced in the hippocampal CA1 and CA2 regions after ischemia, and melatonin reduced laminin degradation and neuronal loss. A TUNEL assay demonstrated that there were TUNEL-positive cells in the hippocampus and the number of TUNEL-positive cells was significantly decreased by melatonin. There was no difference in the ischemia-induced hippocampal neuronal damage between the vehicle- and melatonin-treated groups of MMP-9 knock-out mice.

Significance

These data demonstrate that melatonin suppressed the occurrence of neuronal injury, which might be partly due to its inhibitory effects on MMP-9 in addition to its anti-oxidative effects. MMP-9 may be an important key target of melatonin in neuroprotection against global ischemia.     

Protective effect of glutathione against oxygen-induced growth inhibition of human diploid fibroblasts

The reduced glutathione level in human diploid fibroblasts was increased by the addition of N-acetylcysteine or reduced glutathione ethylester into the culture medium, while it was decreased by the addition of L-buthionine-(R,S)-sulfoximine or diethyl maleate. The hyperbaric oxygen-induced reduction in colony-forming ability was prevented in the N-acetylcysteine- or reduced glutathione ethylester-treated cells, and enhanced in the L-buthionine-(R,S)-sulfoximine- or diethyl maleate-treated cells. The extent of the growth inhibition is well correlated with the cellular glutathione level. It is deduced that glutathione is an important safeguard against the oxygen-induced growth inhibition of human diploid cells.