1.Regulatory effect of dexamethasone on aquaporin-1 expression in cultured bovine trabecular meshwork cells.
Xinchun, XIONG ; Juan, MIAO ; Zulian, XI ; Haijiang, ZHANG ; Bo, HAN ; Yizhen, HU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(6):735-7
To evaluate the effect of dexamethasone on the expression of aquaporin-1 (AQP-1) in cultured bovine trabecular meshwork cells, bovine trabecular meshwork cells were cultured in vitro and reproduced to the third and the fourth generation, then treated with dexamethasone at the concentrations of 5, 25, 50, 250 microg/L respectively for 7 days. Immunohistochemical technique-supervision method was employed to measure, and image analysis system to analyze the expression of AQP-1 in normal cultured bovine trabecular meshwork cells and those treated with dexamethasone. In normal bovine trabecular meshwork cells, the grayscale of AQP-1 positive staining was 167.94 +/- 1.18, while it was 168.92 +/- 0.91, 176.72 +/- 1.80, 180.64 +/- 1.31, 185.64 +/- 1.58 in cells treated with 5, 25, 50, 250 microg/L concentrations of dexamethasone. When the concentration of dexamethasone was higher than 25 microg/L, the expression of AQP-1 was significantly inhibited (P < 0.05). The regulation of AQP-1 expression by dexamethasone in cultured bovine trabecular meshwork cells in vitro may be one of causes that retard the aqueous outflow in glucocorticoid- induced glaucoma.
Aquaporin 1/*biosynthesis
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Aquaporin 1/genetics
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Cells, Cultured
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Depression, Chemical
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Dexamethasone/*pharmacology
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Dose-Response Relationship, Drug
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Immunohistochemistry
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Trabecular Meshwork/cytology
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Trabecular Meshwork/*metabolism
2.Expression profile of aquaporin 1 in patients with menorrhagia.
Chao-chao SUN ; Chun FENG ; Cai-yun ZHOU ; He-feng HUANG
Journal of Zhejiang University. Medical sciences 2007;36(5):433-438
OBJECTIVETo investigate the expression of aquaporin 1 (AQP1) in endometrium of patients with menorrhagia.
METHODSRT-PCR and immunohistochemistry were carried out in twenty women with normal menstrual cycle to confirm the expression of AQP1 in endometrium and locate it. Then 51 women with menorrhagia and 40 women with normal menstrual cycle were included in the study. RT-PCR and immunohistochemistry were used to examine the expression of AQP1.
RESULTAQP1 mRNA was expressed in the human endometrium throughout menstruation cycle, which was mainly located in the endothelia of the capillaries and small blood vessels. Quantification of the immunostaining revealed higher density during secretary phase than that in proliferative phase (P<0.01). The staining intensity and density of AQP1-positive microvessel decreased significantly in simple hyperplasia group (P<0.01) and then gradually increased in complex hyperplasia and atypical hyperplasia groups (P<0.001).
CONCLUSIONDecreased expression of AQP1 may lead to disturbed endometrial vascular remodeling and may be involved in the occurrence of menorrhagia.
Adult ; Aquaporin 1 ; biosynthesis ; genetics ; Endometrium ; blood supply ; metabolism ; Female ; Gene Expression ; Humans ; Immunohistochemistry ; Menorrhagia ; genetics ; metabolism ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction
3.Expression of aquaporin-1 in a guinea pig model of labyrinth destruction.
Journal of Southern Medical University 2006;26(5):664-666
OBJECTIVETo study the expression of aquaporin-1(AQP-1) in the cochlea and endolymphatic sac in guinea pigs with labyrinth destruction.
METHODSChloroform was injected into the tympanum to establish the animal model of labyrinth destruction in guinea pigs, and two-step immunohistochemical method was used to examine the expression of AQP-1 in the cochlea and endolymphatic sac at different time points.
RESULTSAQP-1 expression was fluctuant in accordance with the morphological changes of the spiral ligament fibrocytes in the cochlea: destruction of the spiral ligament cells was followed by AQP-1 expression down-regulation, and AQP-1 up-regulation occurred with the cell regeneration. But no such changes were observed in the endolymphatic sac.
CONCLUSIONAQP-1 may take part in the maintenance of the structural stability of the spiral ligament.
Animals ; Aquaporin 1 ; biosynthesis ; Cochlea ; metabolism ; pathology ; Disease Models, Animal ; Endolymphatic Sac ; metabolism ; pathology ; Female ; Guinea Pigs ; Immunohistochemistry ; Labyrinth Diseases ; metabolism ; Male
4.Expression of aquaporin 1 in human placenta and fetal membranes.
Hui-shu LIU ; Xiao-fei SONG ; Rong-zeng HAO
Journal of Southern Medical University 2008;28(3):333-336
OBJECTIVETo detect aquaporin1 (AQP1) expression in normal human placenta and fetal membranes.
METHODSHuman placenta and fetal membrane specimens were collected from 5 pregnant women with intact membranes undergoing elective cesarean sections at term. Expression and localization of AQP1 was detected by RT-PCR, Western blotting and immunohistochemistry.
RESULTSAQP1 mRNA was detected in the placenta and fetal membranes by RT-PCR, and Western blotting also yielded positive results for the specimens, showing a specific band around 28 kD. Immunohistochemical staining showed AQP1 expression in the vascular endothelial cells and syncytiotrophoblasts of the placenta, epithelial cells of the amnion, and cytotrophoblasts of the chorion.
CONCLUSIONThe presence of AQP1 expression in the placenta and fetal membranes suggest that AQP1 may play an important role in maternal-fetal fluid exchange and amniotic fluid balance.
Adult ; Amnion ; metabolism ; Amniotic Fluid ; metabolism ; Aquaporin 1 ; biosynthesis ; genetics ; Blotting, Western ; Chorion ; metabolism ; Female ; Humans ; Immunohistochemistry ; Placenta ; metabolism ; Pregnancy ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction
5.Influence of estradiol benzoate on fluid reabsorption capacity of rat efferent ductuli.
Quan ZHANG ; Xin-Hua ZHOU ; Xiao-Bin ZHANG ; Qing-Hui ZHOU ; De-Zhi WANG ; Xi-Chang CHEN
National Journal of Andrology 2005;11(11):806-809
OBJECTIVETo study the influence of estradiol benzoate (E2B) on the fluid reabsorption capacity of rat efferent ductuli.
METHODSNewborn male Sprague-Dawley rats were injected subcutaneously with E2B (0.2 mg/5 g body weight), and the histological morphology of efferent ductulus, epithelial ultrastructure, and immunoexpression of AQP-1 were investigated on postnatal day 14, 21, 28, 42 and 56, respectively. Vehicle was given to the controls.
RESULTSAfter exposure to E2B, the lumina of the efferent ductuli dilated greatly (P < 0.05), and the epithelium height decreased significantly (P < 0.01), microvilli of nonciliated cells short and sparse, endocytic apparatus implicated in fluid reabsorption scarce, and with no AQP-1 expression.
CONCLUSIONHigh dosage of E2 B neonatally administrated to rats damages the fluid reabsorption capacity of efferent ductuli.
Absorption ; physiology ; Animals ; Animals, Newborn ; Aquaporin 1 ; biosynthesis ; Body Fluids ; metabolism ; Epithelial Cells ; drug effects ; metabolism ; ultrastructure ; Estradiol ; analogs & derivatives ; toxicity ; Male ; Rats ; Rats, Sprague-Dawley ; Testis ; drug effects ; metabolism ; ultrastructure
6.Screening differentially expressed genes in denucleated K562 cells with restriction display technique.
Min WEI ; Wen-li MA ; Yan-bin SONG ; Xiang-ming MAO ; Ling LI ; Wen-ling ZHENG
Journal of Southern Medical University 2006;26(2):162-165
OBJECTIVETo screen differentially expressed genes in cytochalasin B (CB)-induced denucleated K562 cells by restriction display (RD) technique.
METHODSThe total RNA was isolated and purified from K562 cells before and after CB (10 mug/ml) treatment. The mRNA from both treated and untreated K562 cells were reversely transcribed into cDNA, and the differentially expressed genes were separated using RD technique combined with polyacrylamide gel electrophoresis and sliver staining, followed by cloning, sequencing and homology analysis against GenBank database of these genes.
RESULTSSeven differentially expressed genes were identified in CB-treated cells including aquaporin 1 (AQP1) gene, which was verified to be up-regulated after CB treatment by RT-PCR.
CONCLUSIONAQP1 gene might be in close association with the regulation of denucleation processes and CB-induced proliferation inhibition of K562 cells.
Aquaporin 1 ; biosynthesis ; genetics ; Cytochalasin B ; pharmacology ; Electrophoresis, Polyacrylamide Gel ; Gene Expression Profiling ; methods ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; K562 Cells ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction
7.Rat alveolar type II injured by bleomycin.
Lu KONG ; Zhi-gang WANG ; Jian-zhao NIU ; Ji-feng WANG ; Huan JIN ; Mei-juan YANG ; Ling-qiao WANG ; Bing-hua TANG ; Qiu-ju ZHANG ; Heng-jing TU
Acta Academiae Medicinae Sinicae 2005;27(1):81-86
OBJECTIVETo explore dysfunction mechanism of rat alveolar type II (AT-II) injured by bleomycin (BLM).
METHODSSD rats were injected with a single intratracheal dose of bleomycin or control saline. On day 7, 14, and 28 following intratracheal bleomycin or saline instillation, animals were killed under overdose of 1.5% sodium pentobarbital (0.25 ml/100 g, i.p.) and bronchoalveolar lavage fluid (BALF) from the lung was tested for the activity of pulmonary surfactant (PS) by the Whihelmy Film Balance. Several concentrations of bleomycin stimulated the culture of rat AT-II cells, and surfactant protein (SP) A, B, and aquaporin-1 (AQP) mRNA were analyzed by fluorescent quantitative polymerase chain reaction (FQ-PCR).
RESULTSThe activity of PS and hypoxemia significantly decreased on day 7 and improved on day 14 and completely recovered to normal status on day 28. SP-A, B, and AQP-1 mRNA expression in BLM-stimulated group were significantly lower than those in the control group (P<0.001).
CONCLUSIONBLM-injured AT-II cells decrease the levels of SP-A, B, and AQP-1 mRNA and cause PS dysfunction, resulting in hypoxemia and pneumonedema.
Animals ; Aquaporin 1 ; biosynthesis ; genetics ; Bleomycin ; administration & dosage ; toxicity ; Cells, Cultured ; Dose-Response Relationship, Drug ; Epithelial Cells ; drug effects ; metabolism ; Hypoxia ; chemically induced ; metabolism ; pathology ; Male ; Pulmonary Alveoli ; cytology ; drug effects ; Pulmonary Surfactant-Associated Protein A ; biosynthesis ; genetics ; Pulmonary Surfactant-Associated Protein B ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Time Factors
8.Inhibition effect of epalrestat on rat lens osmotic expansion.
Li-Xia JI ; Ning SHEN ; Cai-Na LI ; Quan LIU ; Yi HUAN ; Zhu-Fang SHEN
Acta Pharmaceutica Sinica 2009;44(10):1107-1111
Epalrestat is the unique aldose reductase inhibitor on the market, which was mainly used for the diabetic neuropathy. Lenses osmotic expansion could be induced by galactose to mimic the pathological process of diabetic cataract in vitro. In present study, we mainly investigated whether epalrestat possesses inhibitory effect on the lens osmotic expansion. The results indicated that epalrestat could not only markedly inhibit rat lens osmotic expansion in vitro, but also significantly reduced the high expression of the osmotic expansion-related genes such as AR and AQP1 in mRNA and protein levels. The findings may provide an important reference to epalrestat in the clinical application for the treatment of diabetic cataract.
Aldehyde Reductase
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antagonists & inhibitors
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biosynthesis
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genetics
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Animals
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Aquaporin 1
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genetics
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metabolism
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Cataract
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etiology
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metabolism
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pathology
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Diabetes Mellitus, Experimental
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complications
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Enzyme Inhibitors
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pharmacology
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Galactose
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antagonists & inhibitors
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Lens, Crystalline
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drug effects
;
metabolism
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pathology
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Male
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Osmosis
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drug effects
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RNA, Messenger
;
metabolism
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Rats
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Rats, Sprague-Dawley
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Rhodanine
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analogs & derivatives
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pharmacology
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Thiazolidines
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pharmacology