Autophagy Mediates MMP-2 Expression in Glaucomatous Trabecular Meshwork Cells
Purpose. To investigate the effect of 3-methyladenine (3-MA) and starvation on the expression of matrix metalloproteinase (MMP-2) in patients with primary open-angle glaucoma. Methods. Primary TM cells were cultured and divided into three groups. The control group was treated with a normal medium, t...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2022-01-01
|
| Series: | Journal of Ophthalmology |
| Online Access: | http://dx.doi.org/10.1155/2022/6026464 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832567765984083968 |
|---|---|
| author | Yan-Ling Xiao Xiao-Rui Wang Kejing Ye Wan-Zhu Chen Bing-Ru Zheng Yi-Hong Huang Yu-Yu Wu |
| author_facet | Yan-Ling Xiao Xiao-Rui Wang Kejing Ye Wan-Zhu Chen Bing-Ru Zheng Yi-Hong Huang Yu-Yu Wu |
| author_sort | Yan-Ling Xiao |
| collection | DOAJ |
| description | Purpose. To investigate the effect of 3-methyladenine (3-MA) and starvation on the expression of matrix metalloproteinase (MMP-2) in patients with primary open-angle glaucoma. Methods. Primary TM cells were cultured and divided into three groups. The control group was treated with a normal medium, the 3-MA group was stimulated with 3-MA, and the starvation group received nutrient depletion by replacing the normal media with Earle’s balanced salt solution. Cellular mRNA and protein were measured at different 3-MA concentrations and starvation time periods. The level of autophagy was accessed by monodansylcadaverine fluorescent staining and expression of specific autophagy-related genes, light chain 3 (LC3), and Beclin1. The effects of 3-MA and starvation on cell proliferation were determined with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay kit. The mRNA and protein expression of LC3-II, Beclin1, and MMP-2 were measured by reverse transcription-polymerase chain reaction and western blot, respectively. Results. Compared to the control group, starvation significantly upregulated LC3-II and Beclin1 in TM cells after 3 h of stimulation, which peaked at 6 h and 9 h, respectively. Increased MDC-labeled cells were also observed. Starvation downregulated the expression of MMP-2. On the contrary, 3-MA suppressed the activation of autophagy, as shown by the marked downregulation of LC3-II and Beclin1. The expressions of MMP-2 were higher in the 3-MA group compared to the control group, reaching a peak at a concentration of 5 mM. Conclusion. Autophagy may be involved in the pathogenesis of POAG via regulating the expression of MMP-2 and, subsequently, the deposition of the extracellular matrix. |
| format | Article |
| id | doaj-art-eade5491934a4c49b32fee8ccf808ab5 |
| institution | Kabale University |
| issn | 2090-0058 |
| language | English |
| publishDate | 2022-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Ophthalmology |
| spelling | doaj-art-eade5491934a4c49b32fee8ccf808ab52025-02-03T01:00:42ZengWileyJournal of Ophthalmology2090-00582022-01-01202210.1155/2022/6026464Autophagy Mediates MMP-2 Expression in Glaucomatous Trabecular Meshwork CellsYan-Ling Xiao0Xiao-Rui Wang1Kejing Ye2Wan-Zhu Chen3Bing-Ru Zheng4Yi-Hong Huang5Yu-Yu Wu6Department of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyPurpose. To investigate the effect of 3-methyladenine (3-MA) and starvation on the expression of matrix metalloproteinase (MMP-2) in patients with primary open-angle glaucoma. Methods. Primary TM cells were cultured and divided into three groups. The control group was treated with a normal medium, the 3-MA group was stimulated with 3-MA, and the starvation group received nutrient depletion by replacing the normal media with Earle’s balanced salt solution. Cellular mRNA and protein were measured at different 3-MA concentrations and starvation time periods. The level of autophagy was accessed by monodansylcadaverine fluorescent staining and expression of specific autophagy-related genes, light chain 3 (LC3), and Beclin1. The effects of 3-MA and starvation on cell proliferation were determined with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay kit. The mRNA and protein expression of LC3-II, Beclin1, and MMP-2 were measured by reverse transcription-polymerase chain reaction and western blot, respectively. Results. Compared to the control group, starvation significantly upregulated LC3-II and Beclin1 in TM cells after 3 h of stimulation, which peaked at 6 h and 9 h, respectively. Increased MDC-labeled cells were also observed. Starvation downregulated the expression of MMP-2. On the contrary, 3-MA suppressed the activation of autophagy, as shown by the marked downregulation of LC3-II and Beclin1. The expressions of MMP-2 were higher in the 3-MA group compared to the control group, reaching a peak at a concentration of 5 mM. Conclusion. Autophagy may be involved in the pathogenesis of POAG via regulating the expression of MMP-2 and, subsequently, the deposition of the extracellular matrix.http://dx.doi.org/10.1155/2022/6026464 |
| spellingShingle | Yan-Ling Xiao Xiao-Rui Wang Kejing Ye Wan-Zhu Chen Bing-Ru Zheng Yi-Hong Huang Yu-Yu Wu Autophagy Mediates MMP-2 Expression in Glaucomatous Trabecular Meshwork Cells Journal of Ophthalmology |
| title | Autophagy Mediates MMP-2 Expression in Glaucomatous Trabecular Meshwork Cells |
| title_full | Autophagy Mediates MMP-2 Expression in Glaucomatous Trabecular Meshwork Cells |
| title_fullStr | Autophagy Mediates MMP-2 Expression in Glaucomatous Trabecular Meshwork Cells |
| title_full_unstemmed | Autophagy Mediates MMP-2 Expression in Glaucomatous Trabecular Meshwork Cells |
| title_short | Autophagy Mediates MMP-2 Expression in Glaucomatous Trabecular Meshwork Cells |
| title_sort | autophagy mediates mmp 2 expression in glaucomatous trabecular meshwork cells |
| url | http://dx.doi.org/10.1155/2022/6026464 |
| work_keys_str_mv | AT yanlingxiao autophagymediatesmmp2expressioninglaucomatoustrabecularmeshworkcells AT xiaoruiwang autophagymediatesmmp2expressioninglaucomatoustrabecularmeshworkcells AT kejingye autophagymediatesmmp2expressioninglaucomatoustrabecularmeshworkcells AT wanzhuchen autophagymediatesmmp2expressioninglaucomatoustrabecularmeshworkcells AT bingruzheng autophagymediatesmmp2expressioninglaucomatoustrabecularmeshworkcells AT yihonghuang autophagymediatesmmp2expressioninglaucomatoustrabecularmeshworkcells AT yuyuwu autophagymediatesmmp2expressioninglaucomatoustrabecularmeshworkcells |