Effects of FOXO4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditions
AIM: To study the effects of FOXO4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditions. <p>METHODS: Human retinal vascular endothelial cells were cultured with high glucose medium. Real-time PCR and Western blot were used to detec...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Press of International Journal of Ophthalmology (IJO PRESS)
2018-12-01
|
| Series: | Guoji Yanke Zazhi |
| Subjects: | |
| Online Access: | http://ies.ijo.cn/cn_publish/2018/12/201812006.pdf |
| _version_ | 1819075244416040960 |
|---|---|
| author | Jun Lu Yu Qin Wen-Wei Xiao Mei Zhang |
| author_facet | Jun Lu Yu Qin Wen-Wei Xiao Mei Zhang |
| author_sort | Jun Lu |
| collection | DOAJ |
| description | AIM: To study the effects of FOXO4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditions. <p>METHODS: Human retinal vascular endothelial cells were cultured with high glucose medium. Real-time PCR and Western blot were used to detect the expressions of FOXO4 within cells; meanwhile, both FOXO4 RNAi lentivirus and control vector lentivirus were infected the retinal vascular endothelial cells cultured in high sugar culture medium. Real-time PCR and Western blot techniques were used to detect the interference efficiency. After collection of supernatant and cells treated with various interferences, the SOD activity, MDA content in the supernatants and ROS level within cells were detected. Flow cytometry was used to determine the changes of cell apoptosis. Western blot was used to detect the expressions of apoptotic protein cleaved Caspase-3 and cleaved Caspase-9 within cells. <p>RESULTS: The expression of FOXO4 was increased in retinal vascular endothelial cells after treatment with high glucose medium. FOXO4 RNAi lentivirus infection reduced the expression level of FOXO4 in retinal vascular endothelial cells under high glucose environmental conditions. By contrast, control vector lentivirus had no effect on FOXO4 expression in cells. High glucose induced elevated levels of ROS in retinal vascular endothelial cells, reduced the activity of SOD in cell culture medium, increased the content of MDA, elevated the rate of apoptosis, and promoted the expressions of cleaved Caspase-3 and cleaved Caspase-9 proteins in cells. After down-regulation of FOXO4 expression, retinal endothelial cells were induced by high glucose, the activity of SOD in the cell culture medium increased, the levels of MDA, ROS, apoptosis, and the levels of cleaved Caspase-3 and cleaved Caspase-9 proteins decreased in cells as compared with those of retinal vascular endothelial cells. Moreover, compared with those did not interfere with FOXO4 expression, there was statistically significant differences(<i>P</i><0.05). <p>CONCLUSION: High glucose induces the expression of FOXO4 in retinal vascular endothelial cells. Knocking-down of FOXO4 expression reduces oxidative stress and apoptosis induced by high glucose medium. |
| first_indexed | 2024-12-21T18:22:20Z |
| format | Article |
| id | doaj.art-eb00b074fbb34d26a19f9d3896d50faa |
| institution | Directory Open Access Journal |
| issn | 1672-5123 1672-5123 |
| language | English |
| last_indexed | 2024-12-21T18:22:20Z |
| publishDate | 2018-12-01 |
| publisher | Press of International Journal of Ophthalmology (IJO PRESS) |
| record_format | Article |
| series | Guoji Yanke Zazhi |
| spelling | doaj.art-eb00b074fbb34d26a19f9d3896d50faa2022-12-21T18:54:30ZengPress of International Journal of Ophthalmology (IJO PRESS)Guoji Yanke Zazhi1672-51231672-51232018-12-0118122146215010.3980/j.issn.1672-5123.2018.12.06Effects of FOXO4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditionsJun Lu0Yu Qin1Wen-Wei Xiao2Mei Zhang3Department of Ophthalmology, Shanghai Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200082, ChinaDepartment of Ophthalmology, Shanghai Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200082, ChinaDepartment of Ophthalmology, Shanghai Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200082, ChinaDepartment of Ophthalmology, Shanghai Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200082, ChinaAIM: To study the effects of FOXO4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditions. <p>METHODS: Human retinal vascular endothelial cells were cultured with high glucose medium. Real-time PCR and Western blot were used to detect the expressions of FOXO4 within cells; meanwhile, both FOXO4 RNAi lentivirus and control vector lentivirus were infected the retinal vascular endothelial cells cultured in high sugar culture medium. Real-time PCR and Western blot techniques were used to detect the interference efficiency. After collection of supernatant and cells treated with various interferences, the SOD activity, MDA content in the supernatants and ROS level within cells were detected. Flow cytometry was used to determine the changes of cell apoptosis. Western blot was used to detect the expressions of apoptotic protein cleaved Caspase-3 and cleaved Caspase-9 within cells. <p>RESULTS: The expression of FOXO4 was increased in retinal vascular endothelial cells after treatment with high glucose medium. FOXO4 RNAi lentivirus infection reduced the expression level of FOXO4 in retinal vascular endothelial cells under high glucose environmental conditions. By contrast, control vector lentivirus had no effect on FOXO4 expression in cells. High glucose induced elevated levels of ROS in retinal vascular endothelial cells, reduced the activity of SOD in cell culture medium, increased the content of MDA, elevated the rate of apoptosis, and promoted the expressions of cleaved Caspase-3 and cleaved Caspase-9 proteins in cells. After down-regulation of FOXO4 expression, retinal endothelial cells were induced by high glucose, the activity of SOD in the cell culture medium increased, the levels of MDA, ROS, apoptosis, and the levels of cleaved Caspase-3 and cleaved Caspase-9 proteins decreased in cells as compared with those of retinal vascular endothelial cells. Moreover, compared with those did not interfere with FOXO4 expression, there was statistically significant differences(<i>P</i><0.05). <p>CONCLUSION: High glucose induces the expression of FOXO4 in retinal vascular endothelial cells. Knocking-down of FOXO4 expression reduces oxidative stress and apoptosis induced by high glucose medium.http://ies.ijo.cn/cn_publish/2018/12/201812006.pdfretinal vascular endothelial cellsoxidative stressapoptosisFOXO4 |
| spellingShingle | Jun Lu Yu Qin Wen-Wei Xiao Mei Zhang Effects of FOXO4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditions Guoji Yanke Zazhi retinal vascular endothelial cells oxidative stress apoptosis FOXO4 |
| title | Effects of FOXO4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditions |
| title_full | Effects of FOXO4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditions |
| title_fullStr | Effects of FOXO4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditions |
| title_full_unstemmed | Effects of FOXO4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditions |
| title_short | Effects of FOXO4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditions |
| title_sort | effects of foxo4 on oxidative stress and apoptosis of retinal vascular endothelial cells under high glucose environmental conditions |
| topic | retinal vascular endothelial cells oxidative stress apoptosis FOXO4 |
| url | http://ies.ijo.cn/cn_publish/2018/12/201812006.pdf |
| work_keys_str_mv | AT junlu effectsoffoxo4onoxidativestressandapoptosisofretinalvascularendothelialcellsunderhighglucoseenvironmentalconditions AT yuqin effectsoffoxo4onoxidativestressandapoptosisofretinalvascularendothelialcellsunderhighglucoseenvironmentalconditions AT wenweixiao effectsoffoxo4onoxidativestressandapoptosisofretinalvascularendothelialcellsunderhighglucoseenvironmentalconditions AT meizhang effectsoffoxo4onoxidativestressandapoptosisofretinalvascularendothelialcellsunderhighglucoseenvironmentalconditions |