The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were stu...The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were studied, and the implication of AQP1 regulation in corticosteroid-glaucoma and the possibility of AS-ODN inhibiting the AQP1 expression were evaluated. The cultured HTM cells in vitro were treated with different concentrations of dexamethasone and transfected with oligonucleotides for 5 days respectively. Then, total RNA and protein of HTM cells were extracted. The changes of AQP1 mRNA and protein were demonstrated qualitatively and quantitatively by RT-PCR and Western blot. Band intensities were detected by imaging analysis. There was a parallel relationship between the results of RT-PCR and those of Western blot. The expression levels of AQP1 mRNA and protein in dexamethasone-treated groups were increased initially and decreased later as dexamethasone concentration was stepped up. In the 0.04 μg/mL and 0.4 μg/mL groups, the levels of AQP1 were higher than in control group （0 μg/mL）. In the 4 μg/ mL and 40 μg/mL groups, the AQP1 expression levels were lower than in control group. AS-ODN could down-regulate the expression of AQP1 mRNA and protein in a dose-dependent manner. At 5 μg/mL, down-regulation efficiency reached the maximum. There was no statistically significant difference in the expression of AQP1 mRNA and protein between all sense oligonucleotides groups and control group. It was suggested that dexamethasone may induce the changes of the AQP1 expression in HTM cells to be involved in the occurrence of corticosteroid-glaucoma. AS-ODN can down-regulate the AQP1 expression in HTM cells to some extent.展开更多
AIM：To investigate the role of Aquaporin-1（AQP-1）in lens epithelial cells（LECs）and its potential target genes.AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparen...AIM：To investigate the role of Aquaporin-1（AQP-1）in lens epithelial cells（LECs）and its potential target genes.AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparency maintenance.Herein,AQP-1 expression in LECs was investigated to evaluate its influence on cell survival in association with its potential role in cataract formation.·M ETHODS：LECs were transfected with lentivirus carrying AQP-1 small interfering RNA（si RNA）.Real-time polymerase chain reaction（PCR）and Western blotting were conducted to detect AQP-1 expression in LECs from different groups.Meanwhile,cell counting kit-8（CCK-8）assay and flow cytometry were performed to measure LEC proliferation and apoptosis,respectively.·RESULTS：AQP-1 expression was significantly reduced in LECs,both at m RNA and protein levels（〈0.05）,after si RNA treatment.Decreased cell viability was detected by CCK-8 assay in LECs with si RNA interference,compared to control cells（〈0.05）.The apoptosis rate significantly increased in cells after si RNA interference（〈0.05）.·CONCLUSION：The decreased cell viability following AQP-1 down regulation is largely due to its induction of apoptosis of LECs.AQP-1 reduction might lead to changes of physiological functions in LECs,which might be associated with the occurrence and development of cataracts.展开更多
AIM： To determine the levels of aquaporin-1（AQP-1） in the lens epithelial cells（LECs） of primary glaucoma and to clarify its correlation with lens thickness.METHODS： This study comprised 64 eyes of 64 patients w...AIM： To determine the levels of aquaporin-1（AQP-1） in the lens epithelial cells（LECs） of primary glaucoma and to clarify its correlation with lens thickness.METHODS： This study comprised 64 eyes of 64 patients with primary glaucoma, who were divided into 3 groups： 25 eyes of 25 patients with acute primary angle-closure glaucoma（APACG）, 19 eyes of 19 patients with chronic primary angle-closure glaucoma（CPACG） and 20 eyes of 20 patients with primary open angle glaucoma（POAG）. This study also included 12 eyes of 12 patients with senile cataract as controls. The levels of AQP-1 in LECs were examined by real-time quantitative polymerase chain reaction（RT-q PCR） and immunohistochemistry. The lens thickness was measured by A-scan ultrasonography. RESULTS： The AQP-1 m RNA levels of LECs were 0.84±0.27, 0.69±0.34, 0.44±0.19 and 0.51±0.21 in APACG, CPACG, POAG and senile cataract group, respectively. The levels of AQP-1m RNA were significantly higher in PACG groups compared with those in senile cataract and POAG group（all P〈0.05）. The immunohistochemistry showed the AQP-1 expression were strong-positive in PACG groups, but weak-positive in senile cataract and POAG group. A positive correlation was found between AQP-1 m RNA levels and the lens thickness（r=0.645, P〈0.001）. CONCLUSION： These findings show that the higher expression of AQP-1 in LECs may contribute to increased lens thickness, which might be associated with the occurrence and development of PACG.展开更多
To evaluate the effect of dexamethasone on the expression of aquaporin-1 （AQP-1） in cultured bovine trabeeular meshwork cells, bovine trabeeular meshwork cells were cultured in vitro and reproduced to the third and ...To evaluate the effect of dexamethasone on the expression of aquaporin-1 （AQP-1） in cultured bovine trabeeular meshwork cells, bovine trabeeular 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, 250μg/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 trahecular meshwork cells and those treated with dexamethasone. In normal bovine trabeeular meshwork cells, the grayseale 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, 250μg/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 trahecular meshwork cells in vitro may be one of causes that retard the aqueous outflow in glueoeorticoid-induced glaucoma.展开更多
Objective To testify the lung injury induced by cardiopulmonary bypass(CPB) in canine model and observe the influence of CPB on the aquaporin 1 (AQP1) mRNA expression in canine lung. Methods 8 mongrel dogs were used t...Objective To testify the lung injury induced by cardiopulmonary bypass(CPB) in canine model and observe the influence of CPB on the aquaporin 1 (AQP1) mRNA expression in canine lung. Methods 8 mongrel dogs were used to perform the cardiopulmonary bypass. The hearts arrested for 90 minutes with mild hypothermia and rebeated for 6 hours. The hemodynamics,the ratio of lung dry weight and wet weight,the plasmic展开更多
Objective:To determine if aquaporin-1 could be detected in cultures of human trabecularshwork cells. Methods: Using primers specific for aquaporin-1, reverse transcription combined withpolymerase chain reaction (RT-PC...Objective:To determine if aquaporin-1 could be detected in cultures of human trabecularshwork cells. Methods: Using primers specific for aquaporin-1, reverse transcription combined withpolymerase chain reaction (RT-PCR) yielded a product and its size with total RNAprepared from the human trabecular meshwork cells. SDS-PAGE and immunoblottingwere also used in this study to detect the specific water channel.Results: The presence of this product and its size (298 base pairs) are consistent withthat of an aquaporin-1 message in these cells. A band of 28 kD in agreement with themolecular size of aquaporin-1 was showed in a film by immunoblotting.Conclusion: The presence of aquaporin-1 in human trabecular meshwork cells, thepredominant cell-type of the primary outflow region of the human eye, suggests that waterchannels may be involved in the movement of aqueous fluid out of the eye. In addition,the existence of aquaporin-1 on cultures of human trabecular meshwork cells provides anin vitro model to study the endogenous expression of aquaporin-1 and its possible role inthe regulation of aqueous outflow.展开更多
文摘The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were studied, and the implication of AQP1 regulation in corticosteroid-glaucoma and the possibility of AS-ODN inhibiting the AQP1 expression were evaluated. The cultured HTM cells in vitro were treated with different concentrations of dexamethasone and transfected with oligonucleotides for 5 days respectively. Then, total RNA and protein of HTM cells were extracted. The changes of AQP1 mRNA and protein were demonstrated qualitatively and quantitatively by RT-PCR and Western blot. Band intensities were detected by imaging analysis. There was a parallel relationship between the results of RT-PCR and those of Western blot. The expression levels of AQP1 mRNA and protein in dexamethasone-treated groups were increased initially and decreased later as dexamethasone concentration was stepped up. In the 0.04 μg/mL and 0.4 μg/mL groups, the levels of AQP1 were higher than in control group （0 μg/mL）. In the 4 μg/ mL and 40 μg/mL groups, the AQP1 expression levels were lower than in control group. AS-ODN could down-regulate the expression of AQP1 mRNA and protein in a dose-dependent manner. At 5 μg/mL, down-regulation efficiency reached the maximum. There was no statistically significant difference in the expression of AQP1 mRNA and protein between all sense oligonucleotides groups and control group. It was suggested that dexamethasone may induce the changes of the AQP1 expression in HTM cells to be involved in the occurrence of corticosteroid-glaucoma. AS-ODN can down-regulate the AQP1 expression in HTM cells to some extent.
基金Supported by the National Natural Science Foundation of China(No.81070715)Innovative Platform Foundation of Fujian ProvinceChina(No.2010Y2003)
文摘AIM：To investigate the role of Aquaporin-1（AQP-1）in lens epithelial cells（LECs）and its potential target genes.AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparency maintenance.Herein,AQP-1 expression in LECs was investigated to evaluate its influence on cell survival in association with its potential role in cataract formation.·M ETHODS：LECs were transfected with lentivirus carrying AQP-1 small interfering RNA（si RNA）.Real-time polymerase chain reaction（PCR）and Western blotting were conducted to detect AQP-1 expression in LECs from different groups.Meanwhile,cell counting kit-8（CCK-8）assay and flow cytometry were performed to measure LEC proliferation and apoptosis,respectively.·RESULTS：AQP-1 expression was significantly reduced in LECs,both at m RNA and protein levels（〈0.05）,after si RNA treatment.Decreased cell viability was detected by CCK-8 assay in LECs with si RNA interference,compared to control cells（〈0.05）.The apoptosis rate significantly increased in cells after si RNA interference（〈0.05）.·CONCLUSION：The decreased cell viability following AQP-1 down regulation is largely due to its induction of apoptosis of LECs.AQP-1 reduction might lead to changes of physiological functions in LECs,which might be associated with the occurrence and development of cataracts.
基金Supported by the Science and Technology Planning Project of Guangdong Province(No.2012B050600032)the Science and Technology Planning Project of Guangzhou(No.1515000176)
文摘AIM： To determine the levels of aquaporin-1（AQP-1） in the lens epithelial cells（LECs） of primary glaucoma and to clarify its correlation with lens thickness.METHODS： This study comprised 64 eyes of 64 patients with primary glaucoma, who were divided into 3 groups： 25 eyes of 25 patients with acute primary angle-closure glaucoma（APACG）, 19 eyes of 19 patients with chronic primary angle-closure glaucoma（CPACG） and 20 eyes of 20 patients with primary open angle glaucoma（POAG）. This study also included 12 eyes of 12 patients with senile cataract as controls. The levels of AQP-1 in LECs were examined by real-time quantitative polymerase chain reaction（RT-q PCR） and immunohistochemistry. The lens thickness was measured by A-scan ultrasonography. RESULTS： The AQP-1 m RNA levels of LECs were 0.84±0.27, 0.69±0.34, 0.44±0.19 and 0.51±0.21 in APACG, CPACG, POAG and senile cataract group, respectively. The levels of AQP-1m RNA were significantly higher in PACG groups compared with those in senile cataract and POAG group（all P〈0.05）. The immunohistochemistry showed the AQP-1 expression were strong-positive in PACG groups, but weak-positive in senile cataract and POAG group. A positive correlation was found between AQP-1 m RNA levels and the lens thickness（r=0.645, P〈0.001）. CONCLUSION： These findings show that the higher expression of AQP-1 in LECs may contribute to increased lens thickness, which might be associated with the occurrence and development of PACG.
文摘To evaluate the effect of dexamethasone on the expression of aquaporin-1 （AQP-1） in cultured bovine trabeeular meshwork cells, bovine trabeeular 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, 250μg/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 trahecular meshwork cells and those treated with dexamethasone. In normal bovine trabeeular meshwork cells, the grayseale 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, 250μg/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 trahecular meshwork cells in vitro may be one of causes that retard the aqueous outflow in glueoeorticoid-induced glaucoma.
文摘Objective To testify the lung injury induced by cardiopulmonary bypass(CPB) in canine model and observe the influence of CPB on the aquaporin 1 (AQP1) mRNA expression in canine lung. Methods 8 mongrel dogs were used to perform the cardiopulmonary bypass. The hearts arrested for 90 minutes with mild hypothermia and rebeated for 6 hours. The hemodynamics,the ratio of lung dry weight and wet weight,the plasmic
基金The study was supported by National Nature Science Foun-dation of China(No.39770789 No.39800163)Doctoral Founda-tion of national teaching association(No.9856)Nature Science Foundation of Guangdong(No.980112)
文摘Objective:To determine if aquaporin-1 could be detected in cultures of human trabecularshwork cells. Methods: Using primers specific for aquaporin-1, reverse transcription combined withpolymerase chain reaction (RT-PCR) yielded a product and its size with total RNAprepared from the human trabecular meshwork cells. SDS-PAGE and immunoblottingwere also used in this study to detect the specific water channel.Results: The presence of this product and its size (298 base pairs) are consistent withthat of an aquaporin-1 message in these cells. A band of 28 kD in agreement with themolecular size of aquaporin-1 was showed in a film by immunoblotting.Conclusion: The presence of aquaporin-1 in human trabecular meshwork cells, thepredominant cell-type of the primary outflow region of the human eye, suggests that waterchannels may be involved in the movement of aqueous fluid out of the eye. In addition,the existence of aquaporin-1 on cultures of human trabecular meshwork cells provides anin vitro model to study the endogenous expression of aquaporin-1 and its possible role inthe regulation of aqueous outflow.
