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Biomedical subjects

Yuping Wang

Publications and source records attributed to Yuping Wang.

At least 37 records · Page 2Linked to original sources
PubMed2005

Placental tissue levels of nonesterified polyunsaturated fatty acids in normal and preeclamptic pregnancies.

OBJECTIVE: We previously reported that maternal plasma levels of nonesterified polyunsaturated fatty acids were decreased in women with preeclampsia as compared to women with normal pregnancies. Polyunsaturated fatty acids of the n-6 and n-3 families are essential dietary fatty acids. The n-6 polyunsaturated fatty acids are involved in inflammatory reactions, whereas n-3 polyunsaturated fatty acids protect against inflammation. METHODS: In this study, we investigated the composition of nonesterified polyunsaturated fatty acids in placental tissue of normal and preeclamptic pregnancies. Linoleic (18:2) and arachidonic (20:4, AA) acids of the n-6 family and linolenic (18:3), eicosapentaenoic (20:5, EPA) and docosahexaenoic (22:6, DHA) acids of the n-3 family were analyzed. CONCLUSIONS: We found that total concentrations of nonesterified polyunsaturated fatty acids were lower in placental tissues from preeclamptic pregnancies than from normal pregnancies. Both n-6 and n-3 polyunsaturated fatty acids were decreased. The decrease in n-6 polyunsaturated fatty acids was due to a decrease in AA. The concentration of linoleic acid was not altered. For n-3 polyunsaturated fatty acids, DHA was decreased, with no change in linolenic acid. The percentage decrease in EPA and DHA was greater than for AA, so the ratio of n-6 to n-3 polyunsaturated fatty acids was two-fold higher in preeclampsia than normal pregnancy. Because the levels of essential fatty acids, linoleic (18:2) and linolenic (18:3), are preserved but levels of metabolites (AA and DHA) are reduced, we speculate that placental oxidative stress and increased conversion of AA to thromboxane are responsible for the decreased concentrations of polyunsaturated fatty acids in preeclampsia.

Adult↗
PubMed2005

Severe acute respiratory syndrome associated coronavirus is detected in intestinal tissues of fatal cases.

OBJECTIVES: A significant percentage of confirmed severe acute respiratory syndrome (SARS) patients experienced gastrointestinal symptoms, and the viral sequence was detectable in the stool of most patients. At present, the knowledge of the pathology of the digestive system in SARS patients is limited. Because a resurgence of the SARS epidemic is constantly possible, there is an urgent need to understand the involvement of the digestive system in this new disease. METHODS: We performed seven SARS autopsies in which samples of alimentary tract and digestive glands were examined with routine pathology, electron microscopy (EM), in situ hybridization (ISH), immunohistochemistry, and real-time polymerase chain reaction (PCR). RESULTS: The main histopathological finding was atrophy of the mucosal lymphoid tissue. A few mucosal epithelial cells and lymphocytes in the intestine were positively stained for coronavirus with ISH. SARS-coronavirus (CoV)-like particles were found in the mucosal epithelial cells under EM and mild focal inflammation was detected in the alimentary tract. One patient who experienced severe diarrhea had pseudomembranous enteritis of the ileum. Fatty degeneration and central lobular necrosis were observed in the liver. No evidence of direct viral infection was found in the esophagus, the stomach, the salivary gland, the liver, or the pancreas. CONCLUSIONS: In addition to the lungs, the gastrointestinal tract is another target of SARS-CoV infection, as the intestinal epithelial cells and mucosal lymphoid tissue are infected. The findings provide possible explanations for the gastrointestinal symptoms and the presence of virus in the stool of SARS patients.

Adult↗
PubMed2005

Cognitive impairment in major depressive disorder revealed by event-related potential N270.

Major depressive disorder (MDD) is a mood disorder that is often accompanied by the impairment of cognitive functions. The aim of this study was to investigate the feasibility of N270 as an index for evaluating the cognitive impairment in MDD patients. Twenty-five patients with MDD diagnosed according to DSM-IV and 25 age matched normal controls performed a matching task while event-related potentials (ERPs) were recorded from their scalp. There were two kinds of stimulus pairs in this study: match condition, the second stimulus (S2) in a pair was identical to the first one (S1); mismatch condition, S2 conflicted with S1 in the color attribute. Subjects were required to press a button in the match condition and to press another button in the mismatch condition. A negative ERP component, N270, which was considered to reflect conflict processing activity in human brain, was evoked by S2 of the mismatch condition. The patient group showed a delayed and smaller N270 than the control group. The prolongation of its peak latency was significant at P3 and P4 electrodes, and the reduction of its peak amplitude was significant at F3, F4, P3 and P4 electrodes. The amplitude of P300 elicited in the match condition was decreased in the patient group at P3 and P4 electrodes, but its latency did not differ from the control group. The results indicate that MDD patients as a group showed cognitive decline. N270 is a sensitive index in revealing cognitive impairment.

Adult↗
PubMed2004

Chemical modification of muscle protein in diabetes.

Levels of glycation (fructose-lysine, FL) and advanced glycoxidation and lipoxidation end-products (AGE/ALEs) were measured in total skeletal (gastrocnemius) muscle and myofibril protein and compared to levels of the same compounds in insoluble skin collagen of control and diabetic rats. Levels of FL in total muscle and myofibril protein were 3-5% the level of FL in skin collagen. The AGE/ALEs, N(epsilon)-(carboxymethyl)lysine (CML) and N(epsilon)-(carboxyethyl)lysine, were also significantly lower in total muscle and myofibril protein, approximately 25% of levels in skin collagen. The newly described sulfhydryl AGE/ALE, S-(carboxymethyl)cysteine (CMC), was also measured in muscle; levels of CMC were comparable to those of CML and increased similarly in response to diabetes. Although FL and AGE/ALEs increased in muscle protein in diabetes, the relative increase was less than that seen in skin collagen. These data indicate that muscle protein is partially protected against the increase in both glycation and AGE/ALE formation in diabetes.

Animals↗
PubMed2004

Evidence of endothelial dysfunction in preeclampsia: decreased endothelial nitric oxide synthase expression is associated with increased cell permeability in endothelial cells from preeclampsia.

OBJECTIVE: The purposes of this study were to examine endothelial nitric oxide synthase expression in endothelial cells and to determine whether the inhibition of endothelial nitric oxide synthase could impair endothelial barrier function in preeclampsia. STUDY DESIGN: Messenger RNA and protein expression for endothelial nitric oxide synthase were examined in endothelial cells that were isolated from normal and preeclamptic pregnancies. Endothelial monolayer permeable response to interleukin-8 stimulation was determined. Normal endothelial cells that were treated with nitric oxide inhibitor were used to test the association of endothelial nitric oxide synthase and endothelial barrier function. Messenger RNA expression for endothelial nitric oxide synthase was determined by reverse transcription-polymerase chain reaction, and protein expression was determined by Western blot analysis. Endothelial permeability was measured by horseradish peroxidase leakage through endothelial cell filters. Interleukin-8 production was measured by enzyme-linked immunosorbent assay. Data were presented as mean+/-SE and analyzed by analysis of variance or nonparametric Mann-Whitney test. RESULTS: Relative messenger RNA expression and protein expression for endothelial nitric oxide synthase were decreased significantly in endothelial cells from preeclampsia compared with cells from normal pregnancies (messenger RNA expression, 0.191+/-0.057 vs 0.508+/-0.061 [P <.01]; protein expression, 0.225+/-0.08 vs 0.786+/-0.098 [P<.01], respectively). Horseradish peroxidase leakage in normal endothelial cells was 0.30+/-0.26 micromol/L (interleukin-8, 1 pg/mL), 3.14+/-2.45 micromol/L (interleukin-8, 5 pg/ml), and 9.08+/-2.69 micromol/L (interleukin-8, 25 pg/mL; P<.01; compared with 0.77+/-0.47 micromol/L [control endothelial cells]). Horseradish peroxidase leakage in preeclamptic endothelial cells was 6.20+/-2.19 micromol/L, 8.44+/-85 micromol/L, and 15.79+/-2.06 micromol/L (P<.05) compared with 5.23+/-1.28 micromol/L, respectively. The ratio of horseradish peroxidase leakage was >7-fold increase in normal endothelial cells, but only a 4-fold increase in preeclamptic endothelial cells in response to interleukin-8 stimulation at 25 pg/mL. The inhibition of endothelial nitric oxide synthase with N(G)-Monomethyl-L-arginine resulted in an increase in interleukin-8-induced endothelial cell permeability. No difference for interleukin-8 production was observed between normal and preeclamptic endothelial cells (1.15+/-0.21 ng/mg protein vs 1.29+/-0.23 ng/mg protein, P>.5). CONCLUSION: Increased endothelial permeability may be associated with decreased endothelial nitric oxide synthase expression and activity in endothelial cells from preeclampsia.

Case-Control Studies↗
PubMed2004

High glucose levels down-regulate glucose transporter expression that correlates with increased oxidative stress in placental trophoblast cells in vitro.

OBJECTIVE: To study glucose transporter expression and oxidative stress in placental trophoblasts under hyperglycemic conditions in vitro. METHODS: Trophoblasts were isolated from term normal human placentas and incubated with Dulbecco's modified eagle medium containing 1000, 2500, and 4500 mg/L glucose for 3 days. At the end of incubation, culture medium was collected. Trophoblast RNA was extracted and mRNA expression of glucose transporters was determined by RNase protection assay. Messenger RNA expression for copper-zinc-superoxide dismutase (CuZn-SOD) was determined by real-time polymerase chain reaction. Lipid peroxide production was determined by measuring malondialdehyde concentration in the culture supernatant. Protein expression of sodium-glucose transporter 2 (SGLT-2) was determined by Western blot analysis. RESULTS: Messenger RNA expression for glucose transporter 1 (GLUT1) and SGLT-2 were reduced in trophoblast cells incubated with 4500 mg/L glucose compared with those incubated with 1000 and 2000 mg/L glucose. mRNA expression of CuZn-SOD was also decreased in trophoblasts incubated with 4500 mg/L glucose. Malondialdehyde production was significantly increased by trophoblasts incubated with 4500 mg/L glucose compared with those by trophoblasts incubated with 1000 and 2000 mg/L glucose (4.69 +/- 0.60 versus 2.10 +/- 0.29 and 2.89 +/- 0.47 nmol/mg protein; P < .01, respectively). CONCLUSIONS: Down-regulation of gene expression of glucose transporters correlates with increased lipid peroxide production and decreased superoxide dismutase expression in placental trophoblasts cultured under hyperglycemic conditions.

Cells, Cultured↗
PubMed2004

Heme oxygenase-1 mediates up-regulation of adhesion molecule expression induced by peroxynitrite in endothelial cells.

OBJECTIVE: Endothelial cell (EC) activation with up-regulation of cellular adhesion molecule (CAM) expression is a pathophysiologic feature in preeclampsia (PE). Enhanced peroxynitrite formation in the vasculature of women with PE was also reported. This study was to test whether EC oxidative stress induced by peroxynitrite could up-regulate EC CAM expression, and whether heme oxygenase-1 (HO-1) has protective effects on this peroxynitrite-induced cellular response. METHODS: Confluent ECs were stimulated with 3-morpholinosydnonimine-HCl (SIN-1, a peroxynitrite generator) alone or combined with Mn(III) tetrakis (1-methyl-4-pyridyl) porphyrin pentachloride (MnTMPyP, a peroxynitrite scavenger) up to 4 hours. EC surface protein expressions for ICAM, VCAM, P-selectin, and E-selectin were measured by colorimetric assay. ECs were also treated with Sn(IV) mesophorphyrin IX dichloride (SnMP, a HO-1 inhibitor) to determine if HO-1 was involved in the increased CAM expression in stressed cells. Protein and mRNA expressions for HO-1 were determined by Western blot analysis and reverse-transcriptase polymerase chain reaction (RT-PCR). Data are presented as the mean +/- SE and analyzed by analysis of variance (ANOVA). RESULTS: Endothelial CAM expressions for VCAM, P-selectin, and E-selectin, but not ICAM, were significantly increased in SIN-1-treated ECs. Protein and mRNA expressions for HO-1 were also up-regulated in cells treated with SIN-1. MnTMPyP blocked both mRNA and protein expressions for HO-1, whereas SnMP only blocked HO-1 protein expression. Both MnTMPyP and SnMP abolished SIN-1-induced up-regulation of VCAM, P-selectin, and E-selectin expression in ECs. CONCLUSIONS: Peroxynitrite-induced EC oxidative stress produces differential effects on CAM expression, which may be mediated by HO-1 regulation. Our results suggest that increased peroxynitrite formation in the maternal vasculature may contribute to the increased CAM expression and enhanced neutrophil-endothelial interaction associated with PE.

Cell Adhesion Molecules↗
PubMed2004

The sequential processing of visual feature conjunction mismatches in the human brain.

To clarify the brain mechanism for multifeature stimulus comparison, subjects matched the features of two serial visual stimuli in pairs. Stimulus pairs were of four categories: C-S-, color same, shape same (match); C-S+, color same, shape different (shape mismatch); C+S-, color different, shape same (color mismatch); C+S+, color different, shape different (conjunction mismatches). Subjects matched the stimuli in three different sessions according to different attention tasks: attending to color (Ac), attending to shape (As), or attending to both color and shape (Acs). A negative one-peak brain potential, N270, was elicited in all the mismatch conditions with amplitude enhanced in the task-relevant mismatch. Negative potential with two peaks, N270 and N400, appeared when attending to the conjunction mismatches concurrently. The two serial negativities in response to attended feature conjunctions might reflect the temporal different stages for processing conjunction mismatches or conflicts.

Adult↗
PubMed2004

Event-related potentials elicited by visual stimulus-duration discrimination tasks.

Event-related potentials (ERPs) were recorded on subjects who discriminated the stimulus duration of two sequentially presented spots (S1 and S2). Four kinds of trials were included: short-duration match (S1 and S2 lasting 300 ms), short-duration conflict (S1 and S2 lasting 300 ms and 600 ms, respectively), long-duration match (S1 and S2 were 600 ms) and long-duration conflict (S1 and S2 were 600 ms and 900 ms, respectively). When S1 was equal to S2, components of P1, N1, P2, N2 and a delayed late positive component were recorded after the onset of S2. The other three negativities were elicited in both short and long duration conflict conditions. The three negative components peaked at 475, 600 and 900 ms in the short-duration condition, and they peaked at 805, 900 and 1200 ms in the long-duration condition. The first negativity was considered a visual mis-match negativity (vMMN), followed by a delayed N270 reflecting the conflict processing for duration in the brain. The last negativity, similar to N270, possibly reflects the second conflict processing activity evoked by stimulus offset.

Adult↗
PubMed2004

Differential expression of vascular endothelial growth factor (VEGF), endocrine gland derived-VEGF, and VEGF receptors in human placentas from normal and preeclamptic pregnancies.

Vascular endothelial growth factor (VEGF) is a potent regulator of placental vascular function. Endothelial dysfunction is a key factor associated with preeclampsia. In this study, we examined expression of VEGF, endocrine gland-derived VEGF (EG-VEGF), VEGF receptors 1 and 2 (VEGFR-1 and VEGFR-2), and neuropilin-1 and -2 (NP-1 and NP-2) in human placentas from women with normal and preeclamptic (PE) pregnancies using quantitative or semiquantitative PCR. We found that total VEGF mRNA expression was increased 2.8-fold (P < 0.05), along with increases in mRNA expression of VEGF121, 165, and 189 (P < 0.05; 1.7-, 1.9-, and 1.8-fold, respectively) in PE vs. normal placentas. Expression of VEGFR-1 mRNA, but not EG-VEGF and the other three VEGF receptors studied, was elevated (P < 0.05) 2.7-fold in PE vs. normal placentas. Protein expression of VEGF and its four receptors was determined using Western blot analysis. For VEGF, two major isoforms (VEGF165 and 189) were detected. For VEGFR-1, VEGFR-2, NP-1, and NP-2, one major band was observed at 180, 235, 130, and 130 kDa, respectively. All of these bands were corresponding to their positive controls. Of these five proteins studied, only VEGFR-1 levels were increased (P < 0.05; 1.7-fold) in PE placentas. The expression of VEGF and the four VEGF receptors was confirmed using immunohistochemistry. They were primarily present in syncytiotrophoblasts and endothelial cells of villous capillaries and large vessels. Thus, together with previous reports that VEGFR-1 mediates trophoblast function and inhibits VEGF-induced angiogenesis and endothelium-dependent vasodilation, these data suggest that the increased VEGFR-1 expression may alter VEGF- mediated function on trophoblast and endothelial cells in PE placentas.

Case-Control Studies↗
PubMed2004

Placental trophoblast-derived factors diminish endothelial barrier function.

Although increased vascular permeability is an important event in the pathogenesis of preeclampsia, the origin of the circulating factor(s) that elicits this endothelial barrier dysfunction is not known. In this study, we use coculture of endothelial cells and placental trophoblast cells to determine whether placental trophoblasts are a potential source of the factor(s) that mediate the increased vascular permeability of preeclampsia. Human umbilical vein endothelial cells grown in Transwell inserts or on coverslips were cocultured with trophoblast cells isolated from normal and preeclamptic placentas or placenta conditioned media. Endothelial cell barrier function was determined by: 1). measurements of electrical resistance and leakage of horseradish peroxidase, and 2). immunofluorescent staining of vascular endothelial-cadherin, pan-cadherin, and occludin. Uterine myometrium endothelial cells were also studied for comparison. We observed the following: 1). electrical resistance was significantly (P < 0.01) decreased (compared with control endothelial cells) in endothelial cell monolayers cocultured with normal trophoblast cells and further reduced in endothelial cells cocultured with preeclamptic trophoblast cells; 2). an increased horseradish peroxidase leakage that was correlated with the decreased electrical resistance in cocultured cells; and 3). disorganized tight junction proteins and an altered distribution of vascular endothelial-cadherin and occludin in monolayers of endothelial cells cocultured with preeclamptic trophoblast cells. Similar responses were noted in uterine myometrium endothelial cells. We conclude that: 1). placental trophoblast cells produce factors that diminish the barrier function of endothelial cells; 2). endothelial tight junctions are more susceptible to factors released from preeclamptic trophoblast cells than from normal trophoblast cells; and 3). these results implicate trophoblast-derived factors in the increased vascular permeability associated with preeclampsia.

Adherens Junctions↗
PubMed2003

Coupling development and elution, a new thin-layer chromatography technique.

Three methods of coupling development and elution were studied in this paper. (1) A new mode of solvent supplementation and eluate collection was developed for descending development. By using a new distributor and collector in descending development, components can be separated and eluted continuously. (2) The same effect can be realized with a slope distributor [Su et al., J. Planar Chromatogr. 14 (2001) 203] and a collector by horizontal development. (3) In-situ elution can be used to treat a developed silica plate, which can elute the separated components to the receptor without scraping them off. These three methods can be used individually, and the in-situ elution can be used with other modes of development.

Chromatography, Thin Layer↗
PubMed2003

Brain potentials elicited by matching global and occluded 3-dimensional contours.

Subjects were instructed to match 3-dimensional forms while event-related potentials (ERPs) were recorded to explore the relationship between visual completion and conflict processing. Sequentially presented paired stimuli (S1 and S2) were identical in condition I, while in condition II the images were of the same contour, but S2 contained an invisible portion, and in condition III S1 and S2 were of different contours. Subjects indicated if stimuli were physically identical or contour similar in two separate sessions. Following the onset of S2, ERP components P100, N150, and LPC were recorded in all conditions. N150 was enhanced in condition II in both sessions, and N270 was elicited in conditions II and III. N150 is related to visual completion, while N270 is related to the evaluation of information discrepancy, belonging to independent systems.

Adult↗
PubMed2003

Event-related potential N270 and its distribution in adults and school-age children.

To compare the pattern and distribution of event-related potential (ERP) N270 in adults and school-age children and to explore the development of conflict processing systems in the human brain, pairs of colored numbers were sequentially presented on a screen to subjects. The subjects were instructed to discriminate whether the color or magnitude of the pairs of numbers were identical (match) or different (conflict), and ERPs were recorded at the same time. In adults, a negative potential peaking at 270ms (N270) was elicited when the second stimulus (S2) conflicted with the first stimulus (S1) in either task relevant or irrelevant conditions. N270 was distributed widely on the scalp in adults. In children, N270 was only elicited by a task-relevant conflict. N270 in the color conflict was distributed bilaterally on the scalp and N270 elicited by the magnitude conflict was found to be on the left central-frontal areas. In school-age children, ERP N270 is elicited under the control of attention. Its distribution on the scalp depends on the attributes of stimulus pairs.

Adult↗
PubMed2003

Event-related potential N270, a negative component to identification of conflicting information following memory retrieval.

OBJECTIVE: N270, an endogenous ERP component of conflict effect, was evoked in previous studies with S1-S2 paradigm. The present experiment is designed to confirm the speculation that this conflict-related negativity could also be elicited by stimulus probes having conflict with a memorized item in a visual post-retrieval comparison task. METHODS: A Sternberg probe-matching paradigm was modified in the present study. The stimuli consisted of a memory set of 3 different items (simple figures) and a retrieval set of 3 probe figures. Subjects matched each probe to its corresponding item in the memory set. The tasks were designed with different conflict loads of no-conflict, low-conflict and high-conflict in the probe retrieval test. RESULTS: Probes of no-conflict elicited a major positive going component, P300, with bilateral parietal distribution. Probes of low- and high-conflict evoked N270, while N430 was elicited only in high-conflict condition. N270 was more negative in high-conflict condition than in low-conflict condition. The N270 was right hemispheric prominent in the low-conflict task and remarkably distributed over the right prefrontal areas. On the other hand, both N270 and N430 were distributed bilaterally on the scalp in the high-conflict task. CONCLUSIONS: The present results demonstrate that the N270 is an index to the conflict identification, while the N430 of the high-conflict task reflects the processing for complex conflicts following probe retrieval. These negativities are related to the processing of conflicts.

Adult↗
PubMed2003

Increased endothelial monolayer permeability is induced by serum from women with preeclampsia but not by serum from women with normal pregnancy or that are not pregnant.

OBJECTIVE: To determine if endothelial monolayer permeability could be altered by serum from preeclampsia (PE). METHODS: Confluent normal endothelial cells (ECs) were incubated with 20% serum from nonpregnant females, normal and PE pregnancies or combined with antioxidant superoxide dismutase (SOD) for 8 hr. Confluent PE ECs were incubated with 20% serum from normal pregnancies. EC barrier function of monolayer permeability was accessed by measuring EC electrical resistance (ER) and the leakage of horseradish peroxidase (HRP) passing through EC filters. Plasma concentrations of IL-8 and lipid peroxides by MDA were also measured. We determined 1) if serum from PE could affect EC permeable function; 2) if antioxidant and serum from normal pregnancies could preserve PE EC barrier function; 3) if lipid peroxides and cytokine IL-8 were increased in PE blood samples. Data are presented as mean+/-SE. ANOVA was used for statistical analysis. A p level less than 0.05 was considered statistically different. RESULTS: 1) ER was significantly decreased and HRP passage was significantly increased in ECs incubated with serum from PE compared to serum from non-pregnant and normal pregnant females (ER: 36.30+/-2.60 vs. 51.30+/-4.00 and 53.90+/-5.80 Omega x cm2, p<0.01; HRP: 0.100+/-0.020 vs. 0.014+/-0.002 and 0.022+/-0.007 DeltaOD470 nm, p<0.01, respectively). 2) ER was improved in PE ECs incubated with serum from normal pregnancies compared to controls, 52.28+/-3.13 vs. 34.50+/-3.80 Omega x cm2, p<0.01. 3) SOD attenuated decreased EC ER induced by PE serum, 55.58+/-3.61 Omega x cm2 (SOD+PE serum) vs. 42.34+/-3.24 (control) and 35.46+/-2.44 (PE serum), p<0.01, respectively. 4) Both MDA and IL-8 concentrations were higher in plasma or serum samples from PE than those in samples from nonpregnancies and normal pregnancies, MDA: 28.65+/-1.45 vs. 22.40+/-1.47 and 25.53+/-0.89 micromol/mL, p<0.01; IL-8: 5.35+/-1.08 vs. 1.69+/-0.47 and 2.28+/-0.73 pg/mL, p<0.05, respectively. Conclusions. 1) Sera from PE but not from nonpregnant women or normal pregnancies increase EC monolayer permeability. 2) Increased lipid peroxides and IL-8 are candidates altering EC barrier function. 3) Antioxidant SOD preserves increased EC monolayer permeability induced by PE serum, suggesting that EC oxidative stress may be associated with altered EC barrier function in preeclampsia.

Adult↗
PubMed2003

Antioxidant superoxide dismutase attenuates increased endothelial permeability induced by platelet-activating factor.

To study the protective effects of the antioxidant superoxide dismutase (SOD) against platelet-activating factor (PAF)-induced endothelial permeability. Endothelial cells (ECs) were isolated from human umbilical veins from normal pregnancies. The first passage (P1) ECs were grown in polycarbonate transwell filters. Confluent ECs were incubated with PAF at concentrations of 2, 5, and 10 microgram/mL for 2 hours or pretreated with superoxide dismutase. Endothelial monolayer permeability was then measured by EC electrical resistance or by the leakage of horseradish peroxide (HRP) passing through filters. Endothelial junctional protein distribution and expression of VE-cadherin and occludin were determined by fluorescent staining of endothelial monolayer and by Western blot analysis. mRNA expressions for VE-cadherin and occludin were determined by reverse transcriptase-polymerase chain reaction. Data are expressed as Omega. cm(2) for electrical resistance and DeltaOD 470 nm for HRP assay and presented as mean +/- standard error of the mean. Analysis of variance was used for statistical analysis. A P value less than.05 was considered statistically significant. Endothelial cell electrical resistance was decreased and HRP leakage was increased in ECs treated with PAF. Intercellular gaps were formed at cell contact regions in ECs treated with PAF, as evaluated by staining of junctional protein VE-cadherin and occludin. The functional changes of the EC barrier and the formation of intercellular gaps induced by PAF were concentration dependent, which could be partially attenuated by pretreatment of ECs with SOD. Total cellular junctional protein expression and mRNA expression of VE-cadherin and occludin were not affected by PAF. Increased EC monolayer permeability induced by platelet-activating factor is associated with disorganization of EC junctional protein distribution of VE-cadherin and occludin. Superoxide dismutase partially attenuated the PAF-induced increased endothelial monolayer permeability, which suggests that oxidative stress might be involved in the process of PAF-induced disturbances of endothelial barrier function.

Antigens, CD↗