Correlation Between Nitric Oxide and Urodynamics in Men With Bladder Outlet Obstruction / 대한배뇨장애요실금학회지

Purpose@#To investigate the correlation between nitric oxide (NO) and urodynamics in men with bladder outlet obstruction (BOO) by analyzing nitric oxide synthase (NOS) in the urothelium. @*Methods@#We prospectively enrolled 25 men who planned to undergo surgical treatment for benign prostatic obstruction and identified as BOO in the preoperative urodynamics. Bladder tissue was taken during surgical prostate resection. Expressions of endothelial NOS (eNOS), inducible NOS (iNOS), and neuronal NOS (nNOS) in the urothelium were analyzed, and their correlation with urodynamic parameters was also assessed in all patients. We also compared the expressions of eNOS, iNOS, and nNOS between BOO with detrusor underactivity (DU) group and BOO without DU group. @*Results@#In all patients, the level of eNOS positively correlated with maximal flow rate and with maximum cystometric capacity (MCC). The level of iNOS positively correlated with MCC. nNOS levels were positively correlated with detrusor pressure at maximal flow and with bladder contractility index in all patients. The level of eNOS, iNOS, and nNOS did not significantly differ between BOO without DU group and BOO with DU group. @*Conclusions@#This study suggests that NO was correlated with bladder dysfunction in men with BOO. Particularly, nNOS may reflect the change in detrusor function.

Analysis of Human Leukocyte Antigen-G Expression in Ovarian Cancer / 대한산부인과학회잡지

OBJECTIVE: Human leukocyte antigen-G (HLA-G) is a nonclassical major histocompatibility complex (MHC) molecule with highly limited tissue distribution that has been proposed to protect tumor cells from natural killer cell lysis. To delineate the potential role of HLA-G in ovarian cancer, we investigated expression patterns of this molecule in human ovarian cancer cell lines and tissues. METHODS: HLA-G expression was determined both at RNA level by reverse transcriptase-polymerase chain reaction (RT-PCR) and protein level by Western blotting and immunohistochemistry using monoclonal antibody against denatured heavy chain of HLA-G, MEM-G1, in 13 ovarian cancer patient tissues and 6 ovarian cancer cell lines (OVCAR-3, SKOV-3, ES-2, PA-1, TOV-112D, TOV-21G). RESULTS: We found mRNA transcripts of different HLA-G isoforms in five of 6 ovarian cancer cell lines (OVCAR-3, SKOV-3, ES-2, TOV-112D, TOV-21G). HLA-G protein was also detected in 5 cell lines that exhibited expression of HLA-G mRNA transcripts. Immunohistochemical analysis of human ovarian cancers revealed expression of HLA-G in eight of 13 tissue samples. CONCLUSION: Our results provide additional clues as to how a tumor can be selected in vitro and in vivo to escape from cytotoxic anti-tumor responses. We suggest that the aberrant expression of HLA-G may contribute to immune escape in human ovarian cancer.

The Effect of Preeclampsia Mother's Serum and Her Neonatal Cord Blood on the Activity of Soluble Adhesion Molecules (sICAM-1) and Apoptosis in Cultured Human Umbilical Venous Endothelial Cells (HUVEC)

PURPOSE: It is proposed that endothelial cell dysfunction, a central pathogenic feature of preeclampsia is caused by circulating unknown factors produced by placenta in the blood of women with preeclampsia. In this study we investigated the effect of such factors in the serum from preeclamptic mother or her neonatal umbilical vein on the activity of sICAM-1 and apoptosis in cultured human umbilical venous endothelial cells. METHOD: Quantitive determinations of sICAM-1 and apoptosis were detected from isolated and cultured HUVEC supernatants which were incubated with serum samples for 48hours. The serum samples were collected from preeclamptic mother and her neonatal cord blood in pairs according to gestational age and were compaired to nonpreeclamptic control groups. RESULTS: sICAM-1 level was significantly higher in the maternal groups compared to corresponding cord groups (P<0.001). The preterm maternal group showed higher level than term maternal group (P<0.001). The preeclamptic preterm mother groups showed elevated level compared to other mother groups and the existence of preterm delivery affects sICAM-1 level more importantly than the existence of preeclampsia. In TUNEL stain preeclamptic preterm mother group showed increased number of apoptotic nuclei compared to other groups. The neonatal cord group of preterm preeclampsia showed no apoptotic body on cultured HUVEC. CONCLUSIONS: Unknown circulating factors in preeclamptic preterm mother activate expression of sICAM-1 and apoptosis in cultured human umbilical venous endothelial cells. But the fetal circulation may not be affected by the factor (s) that lead to disturbed endothelial cell function in women with preterm preeclampsia.

The Effect of Gonadotropins and Cytokines on Human Luteal Cell Apoptosis / 대한산부인과학회잡지

OBJECTIVE: Our object is to evaluate the detailed mechanisms of support and regression of the human corpus luteum. METHODS: To investigate the regulation of luteal function by gonadotropins, cytokines, and prostaglandins, the frequency of apoptosis and expression of Fas, Fas-L, Bcl-2, Bax, p53, caspase-8 were examined in cultured human luteal cells after treatment with various doses of FSH (30, 100, or 300 ng/mL), LH (30, 100, or 300 ng/mL), TGFbeta1 (1, 10, or 100 ng/mL), TNFalpha (1, 10, or 100 ng/mL), or PGF2alpha (1, 10, or 100 ng/mL) for 24 h. Cells were tested for apoptosis by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick end labeling TUNEL) method and cell death detection ELISA. Immunostaning was performed using anti-Fas, Fas ligand, Bcl-2, Bax, and p53 antibodies. RESULTS: Incidence of apoptosis determined by TUNEL method in the group without treatment was 1.7+/-0.5% (0 h), 10.8+/-1.6% (24 h), and 12.9+/-1.2% (48 h), respectively. Spontaneous increase was significant at the latter time points. Significant suppression of incidence of apoptosis was observed with LH and TGFbeta1 (P<0.05). On the other hand, significant induction of incidence of apoptosis was observed with TNFalpha and PGF2alpha (P<0.05). Immunostaining revealed that p53 and Bax expressions after treatment with LH or TGFbeta1 were significantly lower than those without treatment. Bcl-2 and caspase-8 expressions were not significantly affected by any substance addition. Also we found that inductions of apoptosis by TNFalpha and PGF2alpha were not correlated with the expression of Fas, Fas ligand, Bcl-2, Bax, p53 and caspase-8. CONCLUSION: Our results suggest that LH and TGFbeta1 may be involved in the support of luteal function via suppression of apoptosis, and that TNFalpha and PGF2alpha may contribute to luteal regression via its induction in human corpus luteum during early luteal phase. Also, Fas, Fas-L, Bax and p53 may play roles in this apoptosis controlled by LH, and TGFbeta1.

Effects of Magnesium sulfate on the expression of caspase-3 and caspase-9 of cytotrophoblasts under hypoxic condition / 대한산부인과학회잡지

OBJECTIVE: To determine the effect of magnesium sulfate (MgSO4) on the expression of caspase-3 and caspase-9 of cytotrophoblasts in vitro under normal and hypoxic condition as assessed immunoblot analyses. METHODS: Normal cytotrophoblasts were isolated from second trimester placentas and cultured in several physiologically relevant concentrations of MgSO4 under standard tissue culture condition (20% O2) or hypoxic condition (1-2% O2). Cytotrophoblasts apoptosis was estimated by TUNEL staining and by immunoblotting for Caspase-9 and Caspase-3. RESULTS: The apoptotic index was highest in cytotrophoblasts cultured under hypoxic conditions for 36 hour in the absence of MgSO4 and was showed decreasing tendency by the addition of MgSO4 under the same condition. The expression of Caspase-9 did not change under both standard condition and hypoxic condition with increasing MgSO4 concentrations, but the expression of Caspase-3 decreased under hypoxic condition with increasing of MgSO4 concentrations. CONCLUSION: MgSO4 might protect cytotrophoblasts from apoptosis under hypoxic condition.

The Selective Cyclooxygenase-2 Inhibitor Rofecoxib Reduces Chronic Cerebral Hypoperfusion-induced Apoptosis in the Rat Hippocampus

BACKGROUND: Chronic cerebral hypoperfusion induced by permanent occlusion of bilateral common carotid arteries (2VO) in rats caused cognitive deficits and neuronal damage. Cyclooxygenase-2 (COX-2) inhibitor was reported to attenuate both post-ischemic prostaglandin accumulation and neuronal damage. We studied the expression of mRNA of COX-2 in the hippocampus during hypoperfusion and the effectiveness of selective cyclooxygenase-2 inhibitor, rofecoxib, in preventing the neuronal damage of this model. METHODS: Bilateral common carotid arteries of the rat were ligated with silk sutures. The expression of mRNA for COX-1 and COX-2 were detected by the RT-PCR. The first group of animals (n=6) was treated with rofecoxib (10 mg/kg, i.p.) 7 days after operation and the following 7 days. The second group of animals (n=6) was treated with diclofenac sodium (9mg/kg, i.p.) and the third group of animals (n=5) was treated with vehicle (DMSO). TdT-mediated dUTP nick end labeling (TUNEL) technique was performed to estimate delayed cell death. RESULTS: Bilateral carotid artery occlusion (2VO) was shown to induce apoptotic morphology and DNA strand break in hippocampal neurons from 7 days with a peak at 14, 28 days. mRNA of COX-2 appeared in the frontal cortex (14, 28 days) and hippocampus (14, 28, 63 days). Treatment with rofecoxib significantly (p<0.05) attenuated the number of TUNEL-labeled cells in the hippocampus, whereas the cells of the diclofenac treated group were not protected. CONCLUSIONS: We concluded that COX-2 might contribute to cell death of pyramidal cells of the hippocampus of hypoperfusion and selective COX-2 inhibitor, rofecoxib, could prevent the neuronal damage.