ABSTRACT
PURPOSE: To evaluate the effects of trypan blue (TB) on the survival of cultured human trabecular meshwork cells (HTMCs). METHODS: Primarily cultured HTMCs were exposed to 0.05, 0.10 or 0.50% TB for 1, 5 or 30 min. Cellular survival was assessed using the MTT assay and degree of apoptosis was analyzed with flow cytometry using annexin-V/propidium iodide double staining. RESULTS: Long-term exposure or high concentration of TB decreased the survival of HTMCs (p < 0.05). In flow cytometric analysis, exposure to 0.50% TB for 30 min increased the degree of apoptosis (p < 0.05). Commercial TB decreased cell survival after exposure for 5 min and increased the degree of apoptosis after exposure for 30 min (p < 0.05). CONCLUSIONS: TB may cause cellular damage of cultured HTMCs and apoptosis could be the underlying mechanism. In TB-assisted cataract surgery, TB should be used for the shortest time possible and removed completely.
Subject(s)
Humans , Apoptosis , Cataract , Cell Survival , Flow Cytometry , Trabecular Meshwork , Trypan BlueABSTRACT
Background Recent studies have confirmed that sulforaphane (SFN) can activate multiple pathways,and promote the expression of the antioxidants in cells.Thioredoxin (Trx) plays an important role in maintaining the intracellular redox in the steady state.Objective This study was to investigate the effect and mechanism of SFN on Trx expression in bovine trabecular meshwork cells (BTMCs) cultured in vitro.Methods BTMCs were cultured in vitro and identified by morphological evaluation.The third generation of BTMCs were cultured in the medium with 0,10,20 and 30 μmol/L SFN for 30 minutes.Real-time PCR was applied to measure the expression of Trx mRNA in BTMCs.The BTMCs were randomly divided into normal control group,LY294002 group,U0126 group,SFN group,LY294002 +SFN group and U0126+SFN group.The expressions of Nrf2 protein and Trx protein in each group were measured by Western blot.Results The BTMCs was successfully cultured in vitro.The expressions of Trx mRNA were significantly different among the different concentrationss of SFN treatment (F=88.090,P＜0.01).The expressions of Trx protein and Nrf2 protein in the LY294002 +SFN group,U0126 +SFN group and SFN group were significantly higher than those in the normal control group (all at P ＜ 0.01).The expressions of Trx protein and Nrf2 protein in the LY294002+SFN group and U0126+SFN group were significantly higher than those in the SFN group (all at P＜0.01).Conelusions SFN can activate Nrf2 by phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) and mitogen-activated protein kinase (MAPK)/extracellular regulated protein kinases 1/2 (ERK1/2) signaling pathways,which can increase the expression level of Trx in BTMCs cultured in vitro.
ABSTRACT
Background Selective laser trabeculoplasty (SLT) can increase the outflow of aqueous humor and reduce the intraocular pressure of patients with open angle glaucoma,but its mechanism is unknown.To investigate the mechanism of SLT would improve the therapeutic effect of SLT.The aqueous outflow resistance in trabecular meshwork was affected by the extracellular matrix (ECM).Matrix metalloproteinase-3 (MMP-3) and MMP-9 were closely related to ECM degradation in trabecular meshwork.Objective This study was to observe the effects of SLT on the expressions MMP-3 and-9 in human trabecular ceils in vitro.Methods Immortallized human trabecular cells were cultured with pigment particles mixed suspension for 16 hours and incubated overnight.Then the cells were irradiated with Q switch frequency doubling 532 nm Nd:YAG laser to establish SLT-effective cells with the energy of 0.2 mJ,spot diameter of 400 μm and pulse duration of 3 ns.The expressions of MMP-3 mRNA and MMP-9 mRNA in the cells were detected by fluorescence quantitative real time PCR before and 1 hour and 4,8,12,28 hours after exposure of laser.The concentrations of MMP-3 and MMP-9 in the medium were assayed using ELISA 1 hour and 4,8,12,28 hours after exposure of laser and compared between the non-irradiation group and the irradiation group.Results The relative expressing levels of MMP-3 mRNA were 1.00,1.32±0.12,2.08±0.05,2.34±0.04,2.77± 0.05 and 2.49±0.27 in the non-irradiation group and irradiation for 1 hour and 4,8,12,28 hours after exposure of laser SLT,showing a significant difference among the groups (F =15.966,P=0.007),and relative expressing levels of MMP-3 mRNA were significantly higher in various time points after laser irradiation than those of the non-irradiation group (all at P＜0.05).The relative expressing levels of MMP-9 mRNA were 1.00,0.91 ±0.10,1.27 ± 0.07,1.46±0.07,1.69±0.09 and 0.87±0.09 in the non-irradiation group and irradiation for 1 hour and 4,8,12,28 hours after exposure of SLT,which was considerably different among the groups (F =30.526,P =0.005),and significant increased values were seen in the 4,8 and 12 hours after irradiation compared with the non-irradiation group (all at P＜0.05),with highest expression in the irradiation for 12-hour group.The concentrations of MMP-3 and MMP-9 proteins in medium were significantly increased in various time points after laser exposure in comparison with the control group (all at P＜0.05).Conclusions The expressions of MMP-3 and MMP-9 in human trabercuolar cells upregulate and the secretion ability of human trabecular cells to MMP-3 and MMP-9 proteins improves in early stage of SLT in vitro.However,these procedures gradually diminish with the lapse of time.
ABSTRACT
Background At present,a new drug,platelet-derived growth factor-BB (PDGF-BB),as a drug knife for the treatment of glaucoma is under study to expect it directly working on the trabecular meshwork without disrupting the normal physiological structure of anterior chamber angle,clearing the trabecular meshwork aqueous outflow channel so as to achieve the purpose of lowering intraocular pressure.Objective This study aimed to investigate the effect of PDGF-BB on matrix metalloproteinase-2 (MMP-2) expression in cultured bovine trabecular meshwork cells.Methods Trabcular tissue was obtained from fresh bovine eyes,and trabcular meshwork cells were cultured and passaged using explant method.Cultured cells were identified by morphological evaluation and neuronspecific enolase (NSE) staining.The third generation of cells were inoculated to 6-well plate,and different concentrations (0,5.0,12.5,25.0 μg/L) of PDGF-BB was added into the medium for 2 hours.Expression levels (A value) of MMP-2 mRNA (MMP-2 mRNA/β-actin) and protein in the cells were assayed by reverse transcription polymerase chain reaction (RT-PCR) and immunochemistry,respectively.Results Trabcular meshwork cells appeared 5-9 days after cultured.The third generation of cells presented with many process and showed the green influence in cytoplasm.MMP-2 mRNA/β-actin value (A) was 0.127 ± 0.026,0.147 ± 0.045,0.178 ± 0.053 and 0.222±0.062 in the O,5.0,12.5,25.0 μg/L PDGF-BB group,respectively,showing a significant difference among them (F =56.71,P＜0.05),and the MMP-2 mRNA/β3-actin value in the 5.0,12.5,25.0 μg/L PDGF-BB group was elevated in comparison with that of the 0 μg/L PDGF-BB group (all P＜0.05).The expression value (A value) of PDGF-BB protein in the cells was 446.12±13.81,1444.65±54.64,2086.18±73.18,3488.65±25.98 in the 0,5.0,12.5,25.0 μg/L PDGF-BB group,respectively,with a significant difference among the four groups (F=213.12,P＜0.01),and the expression value (A value) of PDGF-BB protein was gradually increased with the ascend of concentration of PDGF-BB(all P＜0.05).Conclusions PDGF-BB can promote the expression of MMP-2 in bovine trabcular meshwork cells in vitro in concentration-dependent manner.
ABSTRACT
Background5-Nitro-2-(3-styrene-acrylic amine) benzoic acid ( NPPB),a chloride channel inhibitor,has a promoting effect on cell apoptosis in myocardial ischemia and reperfusion of domestic rabbit.The CIC chloride channel has been found in the ocular trabecular cells.However,the effect of NPPB on the shape and function of trabecular cells is unclear. Objective This study was performed to investigate the effect of NPPB on the proliferation,cell cycle progression and apoptosis of human trabecular meshwork cells.MethodsThe immortalized human trabcular cell strain was cultured,and logarithmic-phase cells were incubated in 96-well plates at a density of 1 ×106/ml.Different concentrations of NPPB (10,50,100 μ mol/L) were added to the medium,and the MTT assay was used to assess the growth and proliferation of the cells.Flow cytometry was used to evaluate the cell cycle.Then,100 mg/L 5-FU or 100 mg/L 5-FU + 100 μmol/L NPPB was used to induce cell apoptosis,which was assessed by Annexin V-PI.The membrane potential of mitochondria was examined using rhodamine 123 (△ψm).Results After 48 hours of treatment with NPPB,the abosorbency (A value) of the cells was gradually lowered with the increasing dose of NPPB,with significant differences among the 4 groups (F =7.230,P =0.006).Compared with the 10 μmol/L NPPB group,the A values were significantly declined in the 50 and 100 μmol/L NPPB groups (t =1.610,P =0.025 ;t =12.270,P =0.001 ).Forty-eight hours after exposure to NPPB,the percentage of cells in G0/G1 phase was increased and that in the S phase was decreased.The percentages of cells in different phases of cell cycle were significantly different in comparison with their control groups (without NPPB)( P＜0.05 ).Twenty-four and 48 hours after the treatment with 100 mg/L 5-FU,the apoptosis rates of the cells were raised in the 100 mg/L 5-FU group and 100 mg/L 5-FU + 100 μmol/L NPPB group compared to the without NPPB group (t24h =2.130,P =0.023;t48h =4.810,P=0.011 ) ;while that in the 100 mg/L 5-FU+100 μmol/L NPPB group was higher than the 100 mg/L 5-FU group ( t24 h =1.980,P =0.037 ; t48 h =1.290,P =0.028 ),and the mitochondrial membrane potential was lowered ( t24h =1.580,P =0.029 ; F48 h =6.200,P =0.015 ).Conclusions NPPB suppresses the proliferation of human trabecular cells and promotes the cells to enter S phase via the G1/S check point.In addition,ClC might be involved in an anti-apoptosis mechanism through the internal mitochondrial pathway.
ABSTRACT
BackgroundResearches demonstrated that the levels of soluble CD44 (sCD44)molecule in aqueous is significantly higher in primary open-angle glaucomous(POAG) eye than normal eye,but how the sCD44 would affect the expression of apoptosis protein in trabecular meshwork cells is below understanding.Objective The present study was to investigate the effect of sCD44 on the expression of regulatory proteins bcl-2 associated death factor bad in trabecular meshwork cells in the patients with POAG.MethodsHuman scleral tissue with trabecular meshwork were obtained from POAG patients during the surgery.The trabecular meshwork cells were primarily cultured by explant culture method and identified by immunochemistry.The third generation of cells were incubated with free-serum DMEM/F12 medium added differnt dosages of sCD44 (0,1,5,10,25,50 mg/L) for 48 hours.The expression of bad protein in cultured cells was detected using cell counting kit-8 (CCK-8) as the absorbance values at 490 nm(A,90 value),and the bad protein level in cultured cells was assayed by ELISA.ResultsThe cultured cells showed the positive response for laminin ( LM ),neuron specific enolase ( NSE ),fibronectin ( FN ) monoclonal antibodies.The CCK-8 assay showed that the A490 values of the trabecular meshwork cells in 0,1,5,10,25,50 μg/L of sCD44 groups were 0.2460±0.0019,0.1874±0.0015,0.1570±0.0016,0.1302±0.0019,0.1084±0.0018,0.0940±0.0020 respectively with a statistically significant difference among the 6 groups( F =14.922,P =0.000 ),and the A490 values in various dosages of sCD44 groups were significantly lower than the 0 μg/L sCD44 group (P=0.013,0.008,0.011,0.005,0.004).The ELISA assay showed that bad protein levels in 0,1,5,10,25,50 μg/L of sCD44 groups were ( 114.8461 ± 2.9560 ),( 137.8270 ± 2.4259 ),( 161.4194 ± 3.7381 ),( 170.9453 ± 3.2006 ),( 221.2252 ±4.3738 ),( 324.6167±4.4220) ng/L,showing a total difference among them ( F =16.610,P =0.000 ),and the bad protein levels in various dosages of sCD44 groups were significantly lower than the 0 μg/L sCD44 group( P =0.017,0.013,0.008,0.007,0.006).ConclusionssCD44 can contribute to the apoptosis of the trabecular meshwork cells in patients with POAG in certain dose range by regulating the apoptosis regulatory proteins bcl-2 associated death factor bad.
ABSTRACT
BackgroundObstruction of aqueous humor out flow pathway or abnormality of the extracellular matrix( ECM ) of trabecular meshwork cells causes high intraocular pressure. The balance of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases(TIMPs) is critical for the metabolism of ECM. Interleukin1α(IL-1α) can influence outflow of aqueous humor by regulating MMPs level. Objective This study was to investigate the effect of interleukin-1α on the expression of MMP-2,MMP-3 and TIMP-I in cultured swine trabecular meshwork cells.Methods Swine sclera with trabecular meshwork tissue was isolated from 20 swine eyes and cultured with explant cultured method. Cultured cells were passaged and third generation cells were identified by fibronectin ( FN ) and laminin ( LN ) staining. After 24 hours of serum starvation, trabecular meshwork cells treated with IL-1α at the concentration of 10 mg/L were regarded as the IL group,and serum-free culture medium used to treat trabecular meshwork cells was regarded as the control group. The expression of MMP-2, MMP-3 and TIMP-1 proteins in trabecular meshwork cells were detected by immunohistochemistry,and the expression of MMP-2 mRNA, MMP-3 mRNA and TIMP-1 mRNA were detected by RT-PCR. The examination results were compared between the two groups. ResultsThe third generation of cells were positive for FN and LM. Compared with the control group, the expression levels of MMP-3 and TIMP-1 proteins(A value) in trabecular meshwork cells were significantly higher in the IL group than the control group(t=-7. 694,t =-5. 199,P＜0. 05) ,but no obvious difference was found in the expression of MMP-2 between the two groups( t=-2. 365, P＞0.05 ). The higher expression levels in MMP-3 mRNA and TIMP-1 mRNA (A value) in trabecular meshwork cells were seen in comparison with the control group (t =-3. 025,t=-1. 921 ,P＜0. 05). However,similar results were found in the expression of MMP-2 mRNA between the two groups(t =- 1. 173, P＞0.05 ). ConclusionsThe overexpression of MMP-3 and TIMP-1 proteins and their mRNA leads to the imbalance of MMP-3/TIMP-1 and promotes the decomposition of ECM in the trabecular meshwork, and therefore increases aqueous outflow.
ABSTRACT
PURPOSE: To investigate the effect of insulin on the production of nitric oxide (NO) in the trabecular meshwork (TM) cells and the enzymatic synthetic pathway of tetrahydrobiopterin (BH4) synthesis. METHODS: Primarily cultured human TM cells were exposed to 1, 10, and 100 microgram/ml of insulin and 0, 1, 10, 100 and 1000 nM dexamethasone for 3 days. To evaluate the enzymatic pathway of BH4 synthesis, 10 micrometer dexamethasone, 5 mM diaminopyrimidinone, 100 micrometer ascorbic acid, 100 micrometer sepiapterin, or 10 micrometer methotrexate were also co-administered respectively. Cellular survival and NO production were measured with MTT and Griess assay. RESULTS: Insulin enhanced NO production in a dose-dependent manner significantly (p<0.05) without affecting cell viability, whereas dexamethasone inhibited NO production. With co-exposure of insulin, diaminopyrimidinone and sepiapterin inhibited insulin-induced NO production. Ascorbic acid increased NO production independent of insulin and methotrexate did not affect to the action of insulin in NO production. CONCLUSIONS: Insulin increases NO production in TM cells via de novo synthetic pathway for BH4 synthesis. Insulin could be involved in the regulation of trabecular outflow by enhancing NO production in TM cells.
Subject(s)
Humans , Trabecular Meshwork/cytology , Nitric Oxide/biosynthesis , Insulin/administration & dosage , Dose-Response Relationship, Drug , Cells, Cultured , Cell Survival/drug effects , Biopterins/analogs & derivativesABSTRACT
PURPOSE: To identify novel proteins which bind to myocilin, using a yeast two-hybrid screening system. METHODS: A bait plasmid of myocilin was constructed, and then transformed into yeast AH109. The transformed yeast cells were mated with yeast Y187 containing human skeletal muscle cDNA library plasmids in 2 X YPD medium. Mated diploid yeasts were plated on synthetic dropout medium (SD/-Trp-Leu-His and SD/-Trp-Leu-His-Ade). The positive clones grown on the selective medium were amplified, sequenced, and analyzed using the database of GenBank. The expression of selected genes in trabecular meshwork (TM) cells was detected by RT-PCR. RESULTS: The expression of bait protein in yeast was confirmed by Western blot analysis. Extensive screening of a cDNA library led to the selection of twenty-four positive clones which represent eight genes, including six of cytoskeleton associated proteins such as alpha-actin, myosin regulatory light chain 2A, dynactin, syntrophin alpha1, microtubule associated protein 1B, and myosin binding protein C. CONCLUSIONS: Myocilin may interact with cytoskeleton associated proteins in TM cells. Further studies on their interactions will provide functional clues of myocilin.
Subject(s)
Humans , Actins , Blotting, Western , Carrier Proteins , Clone Cells , Cytoskeleton , Databases, Nucleic Acid , Diploidy , Gene Library , Mass Screening , Microtubules , Muscle, Skeletal , Myosin Light Chains , Myosins , Patient Dropouts , Plasmids , Trabecular Meshwork , YeastsABSTRACT
Objective To study the effect of WIN55,212-2 on the expression of PGE2 in cultured bovine trabecular meshwork cells(TMCs) and discuss its mechanism of reducing the intraocular pressure.Methods TMCs were obtained through the cultured tissue,bovine TMCs were prmiarily cultured and subcultured.The third generation bovine TMCs were incubated with different dosages of WIN55,212-2 and the supernatant was collected.The concentration of PGE2 in the medium was measured by ELISA method.PGE2mRNA in TMCs was analyzed by RT-PCR.Result PGE2 in the supernatant and PGE2mRNA in TMCs increased depending on the dosages of WIN55,212-2.Conclusion Certain concentration of WIN55,212-2 may promote PGE2 secretion of TMCs,so as to reduce the intraocular pressure.
ABSTRACT
PURPOSE: This study investigated the role of ascorbic acid on the production of nitric oxide (NO) in the trabecular meshwork (TM) cells. METHODS: After primarily cultured human TM cells were exposed to 1, 10, and 100 micrometer of L-ascorbic acid (LAA), with or without co-administration of 1 mM sodium nitroprusside or 100 micrometer hydrogen peroxide for 48 hr, cellular survival and NO production were measured with MTT and Griess assay, respectively. RESULTS: LAA significantly potentiated NO production in a dose-dependent manner (p< 0.05) without affecting cell viability. LAA increased cell viability after hydrogen peroxide-induced oxidative stress in a dose-dependent manner. LAA enhanced NO production in TM cells and showed a cytoprotective effect against hydrogen peroxide-induced oxidative stress. CONCLUSIONS: LAA might be involved in the regulation of trabecular outflow by enhancing NO production in TM cells.
Subject(s)
Humans , Trabecular Meshwork/cytology , Nitric Oxide/biosynthesis , Dose-Response Relationship, Drug , Cells, Cultured , Cell Survival/drug effects , Ascorbic Acid/administration & dosageABSTRACT
To study the effect of tTG fully phosphorothioated antisense oligodeoxynucleotides (tTG ASDON) on tTG expression in cultured bovine trabecular meshwork cells (BTMCs) in vitro and explore a new treatment alternative for primary open angle glaucoma (POAG), the ASDON1and ASDON2 complementary to the protein codogram region of tTG were designed, synthesized and phosphorothioated according to the secondary structure of tTG. The ASDON1 and ASDON2 were embedded in Lipofectamine and transfected into BTMCs. The untreated group served as negative controls. The expression of tTG in the mRNA and protein level were measured by semi-quantitative RT-PCR and immunohistochemical technique Supervision method respectively. Our results showed that both the mRNA and the protein of tTG with tTG-ASDON1 and tTG-ASDON2 were significantly decreased as compared with that of the controls (P＜0.05). On the other hand, no significant difference was found between the ASDON1 group and the ASDON2 group. It is concluded that the expression of tTG mRNA and protein in cultured BTMC are down-regulated by tTG ASDON. As a result, tTG ASDON may be used for the treatment of POAG through the inhibitory effect on the expression of tTG.
ABSTRACT
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 μg/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 tg/L concentrations of dexamethasone. When the concentration of dexamethasone was higher than 25 μg/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.
ABSTRACT
PURPOSE: To investigate the cellular fate of the mutant myocilins expressed in human trabecular meshwork (TM) cells and their potential cytotoxicity. METHODS: Cultured human TM cells were transduced with adenoviral vectors that expressed green fluorescence protein (GFP) tagged mutant myocilins (G364V, K423E, Y437H, or I477N). The cytopathic effects of the mutant myocilins on the TM cells were verified by protein blot analysis, microscopic examinations, WST-1 cell prliferation assay and yeast two hybrid assay. RESULTS: All of the mutant myocilins examined in the present study accumulated in the endoplasmic reticulum (ER), which was thought to induce ER stress, severe malformed morphology and diminished cell proliferation. In addition, the expression of mutant myocilin could selectively inhibit the secretion of wild-type myocilin. CONCLUSIONS: The dysfunction or decreased number of trabecular meshwork cells due to the cytopathic effect of mutant myocilins may cause diminished turnover of the extracellular matrix of the trabecular meshwork, and thereby increase the resistance of the trabecular outflow, which may be associated with the mechanism of primary open-angle glaucoma.
Subject(s)
Humans , Cell Proliferation , Endoplasmic Reticulum , Extracellular Matrix , Fluorescence , Glaucoma, Open-Angle , Trabecular Meshwork , Two-Hybrid System TechniquesABSTRACT
PURPOSE: To investigate subcellular localizations of wild type myocilin and trunctated (Q368X) myocilin in cultured human trabecular meshwork (TM) cells. METHODS: GFP and GFP tagged truncated myocilin, full-length myocilin, and stromelysin were expressed in TM cells using adenoviral vectors, and their secretory properties were examined by western blotting. To determine the subcellular localizations of myocilins, cellular organelles of the infected TM cells were stained with antibodies or orgenelle specific fluorescent indicators and examined under confocal microscope. RESULTS: Wild type myocilin was expressed as discrete fine vesicles in the perinuclear region of TM cells as well as in ER, but not in microtubules or mitochondria. Colocalization of wild type and truncated myocilin, indicative of in vivo interaction of the proteins, was also observed in cells co-expressing the proteins. Truncated myocilin was found to be accumulated as aggregates in ER, and inhibited the secretion of normal myocilin. CONCLUSIONS: Our result suggests that intracellular accumulation of truncated myocilin may cause a dysfunction of the cells, resulting in alterations in structural compartmentalization of trabecular ECM and obstruction of aqueous outflow.
Subject(s)
Humans , Antibodies , Blotting, Western , Matrix Metalloproteinase 3 , Microtubules , Mitochondria , Organelles , Trabecular MeshworkABSTRACT
PURPOSE: To investigate the cytotoxicities of wild type myocilin and trunctated (Q368X) myocilin in cultured human trabecular meshwork (TM) cells. METHODS: GFP tagged truncated myocilin and DsRED1 tagged wild type myocilin were expressed in TM cells using adenoviral vectors and observed colocalization by confocal microscope. Cytopathic effects in the cells were examined by light microscope and WST-1 cell proliferation assay. RESULTS: Colocalization of wild type and truncated myocilin was observed in cells co-expressing the proteins. Truncated myocilin was found to be toxic to cells, leading to deformed cellular morphology and diminished cell proliferation. CONCLUSIONS: The intracellular accumulation of truncated myocilin exhibited cytotoxicity in trabecular meshwork cells, and eventually resulted in diminished number and dysfunction of trabecular meshwork cells, which might be involved in the pathogenesis of glaucoma.
Subject(s)
Humans , Cell Proliferation , Glaucoma , Trabecular MeshworkABSTRACT
Objective To understand the influence of bradykinin(Bk) on the signaling of prostaglandin E_2(PGE_2) in cultured bovine trabecular meshwork cells(TMCs),and to discuss the mode of interaction between the two signal pathways in TMCs.Methods TMCs were incubated in 1000nmol/L PGE_2 or 1000nmol/L PGE_2 combined with 100nmol/L Bk for 60s.The production of intracellular cAMP was assayed by radioimmunoassay.Results Bk could magnify the stimulation of PGE_2 on cultured TMCs.PGE_2 combined with Bk could cause greater elevation of intracellular cAMP than PGE_2 alone(P
ABSTRACT
OBJECTIVE:To study the effect of dexamethasone (Dex) on expression of aquaporin-1(AQP1) in cultured human trabecular meshwork(HTM) cells METHODS:Reverse transcription combined with polymerase chain reaction(RT-PCR) was used to detect the expression of AQP1 in cultured HTM cells and those treated by Dex RESULTS:The mRNA of AQP1 expressed in normal HTM cells was 1 643?0 354,while 1 577?0 405,1 117?0 443,0 458?0 301,0 267?0 243 in those treated with Dex for 3 days in concentrations of 10-8mol,5?10-8 mol,10-7 mol,5?10-7mol As the concentrations of Dex increased to≥5?10-8mol,the expression of AQP1 was inhibited(P