Biomedical subjects

L Y Wing

Publications and source records attributed to L Y Wing.

At least 19 recordsLinked to original sources

PubMed1998

Ovarian steroids reduced apoptotic death in SV40 temperature-sensitive mutant virus transformed uterine epithelial cells.

Ovarian steroids have been shown to inhibit uterine cell death in vivo. In this study, we investigated whether ovarian steroids regulated cell death in an uterine epithelial cell line transformed with SV40 temperature-sensitive (ts) mutant virus. To assess cell death rate, cells were grown at permissive temperature (34 degrees C) and pulsed with 3H-thymidine. The retention of incorporated radioactivity was then examined after a temperature shift to nonpermissive temperature (40 degrees C) in the absence or presence of estradiol and progesterone. When cells were continuously cultured at 34 degrees C, cell number increased rapidly and most of radioactivity was retained in the attached cells. However, the temperature shift from 34 degrees C to 40 degrees C resulted in a decrease in cell number and radioactivity in attached cells. Estradiol and progesterone attenuated this temperature shift-induced cell death. Morphological examination with Hoechst 33258 staining revealed that the temperature shift increased the percentage of apoptotic death. The treatment of ovarian steroids reduced the extent of apoptotic death. Our studies demonstrated that ovarian steroids could act directly on uterine epithelial cells to reduce apoptotic death in culture.

Animals↗

PubMed1996

Regulation of plasminogen activator inhibitor activity by plasmin in endothelial cells.

The fibrinolytic activity in endothelial cells was regulated by balance of plasminogen activators and plasminogen activator inhibitors. Plasmin can specifically inhibit the biosynthesis of tissue-type plasminogen activator (t-PA), but not plasminogen activator inhibitor, type 1 (PAI-1) in endothelial cells. The PAI activity in the conditioned medium of endothelial cells was low and remained constant in 24 hours. However, the PAI activity in the conditioned medium of the plasmin-pretreated cells increased linearly in 24 hours. Pretreatment with protein kinase C inhibitors, H-7 or staurosporine, partially suppressed the PAI activity induced by plasmin. Pretreatment of endothelial cells with a G-protein inhibitor pertussis toxin resulted in an inhibition of the plasmin-induced PAI activity. The phospholipase A2 inhibitor mepacrine specifically eliminated the effect of plasmin stimulation on PAI activity. Cyclooxygenase and lipoxygenase inhibitors also partially inhibited the plasmin-stimulated PAI activity in endothelial cells. All these inhibitors did not affect the biosynthesis of the PAI-1 antigen in the presence or absence of plasmin. The results indicate that plasmin increased the PAI activity of endothelial cells via pathways in which protein kinase C, G protein, and phospholipase A2 may be involved.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1995

Changes of protein kinase C subspecies in staphylococcal enterotoxin-B-induced thymocyte apoptosis.

Our previous studies demonstrated that intravenous administration of staphylococcal enterotoxin B (SEB) to BALB/c mice resulted in thymocyte apoptosis. In the present study, we evaluated the role of protein kinase C (PKC) in thymocyte apoptosis induced by SEB. Our results showed that the level of protein phosphorylation in the thymocytes was reduced after the in vivo SEB treatment for 24 h. The activity of classical PKC subspecies was decreased in both cytosolic and membrane fractions of thymocytes following SEB administration. The lowest level of PKC activity was reached by 24 and 48 h, then was recovered gradually after 72 h. Furthermore, the mRNA expression of PKC-beta and, to a much less extent, PKC-alpha, but not PKC-sigma, in thymocytes was reduced by SEB. The decrease of mRNA level of PKC-beta showed good correlation with the pattern of PKC activity. These results provide direct evidence showing the changes in PKC subspecies mRNA expression during the process of apoptosis.

Animals↗

PubMed1994

Diacylglycerol formation induced by human plasmin in cultured endothelial cells.

Treatment of cultured bovine carotid artery endothelial cells with 10(-7) M plasmin increased the cellular diacylglycerol which was determined by the formation of [3H]palmitate-labeled diacylglycerol and diacylglycerol mass. Upon the stimulation with plasmin, a gradual increase in diacylglycerol formation was observed within 20 min then slightly declined. The maximal effect during the 1-h time course study was 45 and 55% increases in [3H]palmitate-labeled diacylglycerol and diacylglycerol mass, respectively, at 20 min after plasmin treatment. Formation of phosphatidylethanol was also studied in [3H]palmitate-prelabeled cells in the presence of ethanol. Treatment with plasmin for 20 min induced a significant 45% increase in phosphatidylethanol formation. The present results indicate that the plasmin-induced diacylglycerol formation in endothelial cells was at least in part mediated through the phospholipase D activation.

Animals↗

PubMed1994

Regional differences in prostaglandin production rates among porcine intrathoracic vessels.

To investigate the regional variability in intrathoracic vascular prostaglandin (PG) synthesis, we obtained vessel segments from porcine coronary artery (COA), thoracic aorta (AT), common carotid artery (CRA), pulmonary artery (PA), pulmonary vein (PV), and inferior vena cava (IVC). Vascular production rates of 6-keto-PGF1 alpha (an indicator for PGI2), PGF2 alpha, and PGE2 were measured both in unstimulated state and in arachidonic acid-stimulated state using immunosorbent assays. Our results indicated that PGI2 production rate in all vessel segments decayed with time after vessel dissection. In all vessel segments tested under unstimulated conditions, PGI2 production rates were about one order of magnitude higher than PGF2 alpha and PGE2 production rates of the same specimens. Results from unstimulated, 1.5 hr pre-incubated specimens indicated that i) PGI2 production rates in COA, AT, and PV were greater than those in CRA, PA, and IVC; ii) PGF2 alpha production rates from the same specimens were higher in PV than in AT, CRA, and IVC, while these in PA were higher than in IVC; and iii) PGE2 production rates from the same specimens were not significantly different from one another. Arachidonic acid added at about 1.5 hr after vessel harvest stimulated the PGI2 and PGF2 alpha synthesis rates by 3 to 15 folds. However, this arachidonic acid treatment caused 70 to 300-fold increases in PGE2 production rates, reaching levels comparable to PGI2. All three prostanoid production rates under stimulated conditions were also variable among different intrathoracic vessels. Although either physiological gas concentrations or local hemodynamic conditions alone can partially explain our results, which physiological parameter(s) actually causes these regional differences remains to be verified.

Animals↗

PubMed1993

Human plasmin induces a receptor-mediated arachidonate release coupled with G proteins in endothelial cells.

Treatment of cultured bovine carotid artery endothelial cells with 10(-7) M plasmin increased arachidonate release coupled with the increase in prostacyclin production. The stimulatory effect of plasmin on arachidonate release could be divided into the early and late phases according to its calcium dependency and pertussis toxin sensitivity. The early phase of plasmin-induced arachidonate release was a calcium-dependent and pertussis toxin-sensitive response, which was observed within 20 min after plasmin treatment. The late phase was a calcium-independent and pertussis toxin-insensitive response, which was induced gradually from 20 to 60 min. Induction of the early phase of plasmin's effect required both the lysine binding and catalytic sites in plasmin molecule because it was inhibited either by the binding antagonist tranexamic acid or by the serine protease inhibitor aprotinin. Guanosine 5'-O-(2-thiotriphosphate) potentiated the effect of plasmin in permeabilized or nonpermeabilized cells, indicating that the early phase effect was mediated by a pertussis toxin-sensitive guanosine 5'-triphosphate (GTP)-binding protein. The late phase of plasmin's effect was due to the catalytic activity because it was inhibited by aprotinin but not by tranexamic acid. Microplasmin structurally having the catalytic sites induced a similar late phase effect. Plasmin did not elicit the metabolism of phosphatidyl polyphosphoinositides. These studies demonstrate that the activation of phospholipase A2, which results in arachidonate release, in the early phase of plasmin's effect is a receptor-mediation via GTP-binding protein that is not coupled through phospholipase C activation.

Animals↗

PubMed1993

Acetylpolyamines decrease blood pressure, [Ca++]i and isometric force of vascular smooth muscle.

Polyamines are polycations in cells and acetylation is the first step in their intracellular metabolism. We investigated the effects of the acetylated polyamines on arterial blood pressure and vascular reactivities in rats. Acetylspermine and acetylspermidine, administered at concentrations ranging from 12.5 to 50 mmol/kg b.wt., both induced a transient decrease in mean arterial blood pressure. The magnitudes of the hypotensive effects of these acetylpolyamines and polyamines were in the order of spermine > acetylspermine > acetylspermidine = spermidine. Pretreatment of rats with calcium diminished polyamine-induced hypotensive effects. The effects of spermine and acetylspermine on isolated vascular smooth muscle were examined in rat aortic rings and tail artery strips. Both compounds relaxed precontracted arterial preparations, and this relaxation could be counteracted by increasing extracellular calcium concentration. Tail artery strips were more sensitive to acetylspermine when compared to aortic rings. In tail artery strips preloaded with the bioluminescent protein aequorin, both spermine and acetylspermine caused a concomitant decrease in intracellular calcium and isometric force activated by 36 mM of KCl. These results demonstrate clearly that acetylspermine and spermine alike decrease intracellular calcium concentration of vascular smooth muscle, which is likely to account for the relaxation of vasculature. The relaxation of smooth muscle in the vascular wall in turn might lead to decreased arterial blood pressure.

Animals↗

PubMed1992

Plasmin and the regulation of tissue-type plasminogen activator biosynthesis in human endothelial cells.

Plasmin inhibited the biosynthesis of tissue-type plasminogen activator (tPA) antigen by human umbilical vein endothelial cells (HUVEC) in a dose-dependent manner. The amount of tPA antigen found in the 24-h conditioned medium of cells treated with 100 nM plasmin for 1 h was 20-30% of that in the control group. However, in contrast to tPA, such treatment led to a 3-fold increase in plasminogen activator inhibitor (PAI) activity, whereas the amount of PAI type 1 antigen was unchanged. The effects of plasmin on HUVEC were binding- and catalytic activity-dependent and were specifically blocked by epsilon-aminocaproic acid. Microplasmin, which has no kringle domains, was less effective in reducing tPA antigen biosynthesis or enhancing PAI activity in HUVEC. Kringle domains of plasmin affected neither tPA antigen nor PAI activity of the cells. Other proteases including chymotrypsin, trypsin, and collagenase at comparable concentrations did not have a significant effect on the biosynthesis of tPA antigen or PAI activity of HUVEC. Thrombin stimulated the biosynthesis of tPA and PAI-1 antigens by HUVEC. Thrombin also stimulated an increase in the protein kinase activity in HUVEC, whereas plasmin inhibited the protein kinase activity of the cells. It is possible that plasmin regulates the biosynthesis of tPA in HUVEC through the signal transduction pathway involving protein kinase.

Aminocaproic Acid↗

PubMed1992

Role of polyamines in regulation of uterine blood flow of ovariectomized rats.

Polyamines have been shown to relax several types of smooth muscle including vasculature. In the estrogen-treated uterus, uterine blood flow and polyamine levels in the uterus are increased. The relaxant effect of polyamines on blood vessels suggest that polyamines may act on uterine vasculature to cause uterine hyperemia. In this study, we examined the roles of polyamines in regulating uterine blood flow. Ovariectomized rats were administered with polyamines or estrogen. The uterine blood flow was then measured by employing radioactive microspheres. The direct injection of polyamines into systemic circulation caused a 3-fold increase in uterine blood flow within 30 seconds. The polyamine-induced uterine hyperemia was counteracted by increasing extracellular calcium concentration. When rats were treated with estradiol, uterine blood flow increased. However, the treatment with alpha-difluromethylornithine, a polyamine synthesis inhibitor, did not attenuate the estrogen-induced uterine blood flow. Our results suggest that polyamines may affect uterine blood flow via antagonizing the entry of extracellular calcium. However, the detailed mechanisms via which polyamines involved in estrogenic stimulation of uterine hyperemia may require further studies.

Animals↗

PubMed1992

Chemotactic and mitogenic activities of granulosa cells in developing follicles.

Chemotactic and mitogenic activities of granulosa cells in developing follicles were studied. Immature rats were subcutaneously injected with 20 IU of pregnant mare's serum gonadotrophin and killed at various intervals after injection. The ovaries were removed and granulosa cells were isolated and cultured in a serum-free medium supplemented with insulin, transferrin and hydrocortisone. Chemotactic and mitogenic activities in the conditioned medium were determined. Our results demonstrated that in addition to mitogenic activity, chemotactic activity was also expressed in the conditioned medium of granulosa cells. Both activities increased with the maturity of follicles. A gel filtration analysis revealed that there were two peaks showing both mitogenic and chemotactic activities with a molecular size smaller than 5000. These peaks had various sensitivities to heat and trypsin treatment. In addition, the active component of both peaks was organic solvent-extractable. A thin-layer chromatography analysis indicated that the lipid component was not prostaglandin, estradiol or hydrocortisone.

Analysis of Variance↗

PubMed1992

Cytoprotective effect of reduced glutathione in hydrogen peroxide-induced endothelial cell injury.

The kinetic effects of hydrogen peroxide (H2O2) on cultured endothelial cells isolated from bovine carotid artery were studied. The cytoprotective effects of glutathione (GSH) on H2O2-induced cell injury were also investigated. H2O2-induced a dose- and time-dependent cell injury in cultured endothelial cells. H2O2-induced cell injury was blocked by simultaneous treatment by catalase, but not by superoxide dismutase. H2O2 also induced endogenous PGI2 biosynthesis, and the maximum PGI2 production was reached after 1 h treatment. Stimulation of PGI2 production was parallel with arachidonate release from H2O2-treated cells. However the prostaglandin biosynthesis enzyme activity in cells was inhibited by H2O2 treatment. When the cells were treated with GSH, the intracellular GSH reached a plateau after 3 h treatment. Both H2O2-induced cell injury and PGI2 production were significantly inhibited by the 3 h pretreatment with GSH. The cytoprotective effect of GSH was completely inhibited by buthionine sulfoximine which is a specific inhibitor of gamma-glutamylcysteine synthetase. The results indicate that the cytoprotective effect of GSH on H2O2-induced cell injury in cultured bovine carotid artery endothelial cells depends on the increase in intracellular GSH content.

Animals↗

PubMed1992

Antigen-specific tachycardia and hypotension in rodents.

Tachycardia and hypotension, two cardiovascular responses to anaphylaxis, were specifically induced by antigen in mice and rats, respectively. Intravenous injection of poly (Glu60Ala30Tyr10) (GAT) elicited tachycardia within 30-40 sec in GAT-primed B6 mice. Moreover, a minute amount of GAT (0.2 micrograms) was enough to sensitize the mice to subsequent GAT-induced tachycardia. Challenging doses ranging from 100 ng to 500 micrograms. could elicit tachycardia. The kinetics of tachycardia induction was different from that of antibody production or delayed-type hypersensitivity. Tachycardia was induced from day 6 after immunization, while delayed-type hypersensitivity developed as early as day 4, and anti-GAT antibodies were undetectable on day 6 and would not reach a maximum until day 8. Specific antigen-induced hypotension was also observed in rats. Furthermore, cardiovascular changes in both species could be passively transferred by heat-treated (56 degrees C, 30 min) sera from immunized animals. These benchmarks of antigen-induced cardiovascular changes in mice or rats could be used as models to study the immune control of cardiovascular changes in anaphylactic responses.

Amino Acid Sequence↗

PubMed1990

Differential effects of sex steroids on uterine and renal ODC activity in ovariectomized rats.

Many hormones are known to induce the activity of ornithine decarboxylase (ODC), the first and rate-limiting enzyme for polyamine biosynthesis, in their target tissues. Using ovariectomized rats, we have compared the effects of sex hormones on ODC activity in the uterus and the kidney which contain estrogen and androgen receptors. The results show that 1) both estrogen and androgen stimulate renal ODC activity, 2) estrogen but not androgen effectively increases ODC activity in the uterus, 3) estrogen at higher dosage can stimulate renal ODC activity to an extent similar to that in the uterus, 4) daily treatment with estradiol for 5 days results in the desensitization of uterine ODC activity, but not that of renal ODC activity to the hormonal stimuli. Although both uterus and kidney are targets of sex hormones, our results indicate that estrogen and androgen have differential influences on the ODC activity in these two organs.

Androgens↗

PubMed1988

Differential responses of sex steroid target tissues of rats treated with 4-hydroxyandrostenedione.

4-Hydroxyandrostene-3,17-dione (4-OHA) inhibits ovarian aromatase activity and causes regression of carcinogen-induced hormone-dependent mammary tumors in rats. Although estrogen levels were reduced, LH levels did not increase nor did uterine weight decline in 4-OHA-treated animals. These findings are in contrast to those in animals deprived of estrogen by ovariectomy. The possible direct action of 4-OHA on gonadotropin secretion and uterine growth was, therefore, investigated in ovariectomized rats not treated with the carcinogen. Treatment with 4-OHA for 2 weeks prevented regression of the uterus and the increase in gonadotropin secretion in ovariectomized rats in a dose-dependent manner. The effect on gonadotropin secretion of 4-OHA at 50 mg/kg.day was similar to that of dihydrotestosterone at 0.5 mg/kg.day and could be completely antagonized by administration of the antiandrogen flutamide. The stimulation of uterine growth by 4-OHA was also blocked by flutamide, but not by the antiestrogen enclomiphene. The trophic action of 4-OHA at 50 mg/kg.day was equivalent to that of 1.8 mg/kg.day dihydrotestosterone. Furthermore, treatment with 4-OHA caused a reduction in uterine estrogen receptor and progesterone receptor levels. The reduction in uterine estrogen and progesterone receptor levels was also counteracted by the concomitant injection of flutamide, but not by enclomiphene. The results suggest that in the rat 4-OHA has multiple actions on sex steroid target tissues in addition to inhibition of aromatase. The effects appear to be related to the androgenic rather than estrogenic activity of the compound. Inhibition of gonadotropins may help maintain reduced ovarian estrogen secretion and contribute to the antitumor activity of this compound.

Androstenedione↗

PubMed1988

Effect of estradiol on the activities of ornithine decarboxylase and S-adenosyl-methionine decarboxylase in tissues of ovariectomized rats.

Polyamines play an important role in cellular growth and differentiation. Ornithine decarboxylase (ODC) and S-adenosyl-methionine decarboxylase (SAMD) catalyze the rate-limiting steps of polyamine formation. Many hormones have been known to affect these decarboxylase activities in their target tissues. This study investigated the effect of estradiol (E2) on the activities of ODC and SAMD and on the growth of three estrogen receptor-containing organs: uterus, kidney and liver. Two weeks after ovariectomy, Sprague-Dawley rats received a subcutaneous injection of E2 (10 micrograms/rat). The ODC activity and organ weight were determined at various intervals after E2 administration. The E2 treatment increased the weight of uterus but not that of liver and kidney. In the uterus, ODC activity showed biphasic increases at 6 h and 16-24 h respectively after E2 injection. Renal ODC activity increased gradually and peaked at 18-24 h whereas hepatic ODC activity remained unchanged. Daily injection of E2 over a 3-day period provoked an increase of uterine weight, uterine ODC and SAMD activity. The same treatment also increased renal ODC activity, however it had no effect on the renal SAMD activity and the kidney Our results indicate that E2 has different effects on the activities of ODC and SAMD in three organs tested, and the increase of ODC activity is not necessarily correlated with the tissue growth.

Adenosylmethionine Decarboxylase↗

PubMed1986

Aromatase inhibitors and the treatment of breast cancer.

Inhibition of aromatase to reduce estrogen production by peripheral and ovarian tissue could be a useful approach to treating hormone-dependent breast cancer. Several C19, 17 keto steroids have been identified as aromatase inhibitors. The most potent of these cause rapid competitive inhibition followed by enzyme inactivation. Injections of the compounds caused inhibition of peripheral aromatization in monkeys. In rats, these treatments result in inhibition of ovarian aromatase and estrogen secretion, accompanied by marked regression of carcinogen(DMBA or NMU)-induced mammary tumors. To date, 60 postmenopausal patients with advanced metastatic breast cancer and unselected for the presence of estrogen receptors have been treated with once weekly injections of 4-OHA. The mean estradiol level measured in 14 patients was significantly reduced to 36% of pretreatment values after 1 month and remained at this level for up to 4 months. There was no effect of treatment on gonadotropin levels. Although all patients had relapsed from previous therapy, complete or partial tumor regression occurred in 30% of patients while 15% had static disease. The results indicate that in these patients the responses are due to inhibition of peripheral aromatization and that 4-OHA may be of value in treating postmenopausal breast cancer.

Adult↗

PubMed1986

Aromatase inhibitors and their potential clinical significance.

Estrogen biosynthesis occurs not only in reproductive tissues of the female but also in such diverse sites as testes, adipose and muscle. Our rationale for the clinical use of aromatase inhibitors is that compounds interacting with aromatase in all tissues could provide both selective and effective inhibition of estrogen production. The most potent inhibitor identified by us to date is 4-hydroxyandrostene-3,17-dione (4-OHA). This compound causes rapid competitive inhibition followed by irreversible inactivation of aromatase. Treatment of rats with 4-OHA results in inhibition of ovarian aromatase and estrogen secretion, accompanied by marked regression of carcinogen induced mammary tumors. Using rhesus monkeys, marked inhibition of peripheral aromatization by 4-OHA was also demonstrated. The first clinical study with a selective aromatase inhibitor was recently carried out using once weekly injections of 500 mg 4-OHA in 60 postmenopausal patients with advanced metastatic breast cancer and unselected for the presence of estrogen receptors. The mean serum estradiol level reduced to 36% of pretreatment values for at least 4 months. No effect of treatment on gonadotropin levels occurred indicating that the reduction in estrogen levels was due to inhibition of peripheral aromatization. In spite of the fact that all patients had relapsed from previous therapy, complete or partial tumor regression occurred in 30% of patients while 15% had static disease. Although the optimum dose of 4-OHA has not yet been established, this aromatase inhibitor appears to be of value in treating postmenopausal breast cancer and may be beneficial in other diseases associated with estrogens.

Androstenedione↗

PubMed1985

Effects of aromatase inhibitors, aminoglutethimide, and 4-hydroxyandrostenedione on cyclic rats and rats with 7,12-dimethylbenz(a)anthracene-induced mammary tumors.

4-Hydroxyandrostenedione (4-OHA) is a more potent and specific inhibitor of aromatase (estrogen synthetase) than aminoglutethimide (AG). The two inhibitors were compared in rats with 7,12-dimethylbenz(a)anthracene-induced, hormone-dependent tumors and in normal cyclic rats treated for 4 and 2 weeks, respectively. Ovarian estradiol levels and aromatase activities were not consistently reduced, and tumors regressed in only two of eight rats treated with AG. In animals treated with 4-OHA or 4-OHA:AG, the total tumor volume, estradiol levels, and aromatase activity decreased by greater than 70%. Ovarian weights and plasma luteinizing hormone (LH) levels were also reduced by 4-OHA but increased by AG. Uterine weights were not altered by AG treatment but were increased by 4-OHA. Similar but more consistent results were obtained with these treatments in normal, cyclic rats. In ovariectomized rats, AG had no effect, whereas 4-OHA decreased LH levels and increased uterine weights. The results suggest that, although AG reduces ovarian estrogen secretion by aromatase inhibition, this may lead to an increase in LH secretion. Increased LH may promote ovarian growth and aromatase synthesis, counteracting the inhibitory action of AG to some extent. 4-OHA which inactivates aromatase may also prevent new enzyme synthesis by directly inhibiting gonadotropins. This would result in more effective reduction in ovarian estrogen production by 4-OHA than AG during long-term treatment.

9,10-Dimethyl-1,2-benzanthracene↗