Protective Effects of One 2,4-Dihydro-3H-Pyrazol-3-one Derivative against Posterior Capsular Opacification by Regulation of TGF-β2/SMADs and Non-SMAD Signaling, Collagen I, and Fibronectin Proteins
Many elderly individuals frequently experience cataracts that interfere with vision. After cataract surgery, the left lens epithelial cell (LEC) exhibited fibrosis and posterior capsule opacification (PCO). Sometimes, there is a need for a second surgery; nevertheless, people try other methods, such...
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MDPI AG
2022-10-01
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| Series: | Current Issues in Molecular Biology |
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| Online Access: | https://www.mdpi.com/1467-3045/44/10/343 |
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| author | Chun-Ching Shih Chia-Yi Lee Fung-Fuh Wong Cheng-Hsiu Lin |
| author_facet | Chun-Ching Shih Chia-Yi Lee Fung-Fuh Wong Cheng-Hsiu Lin |
| author_sort | Chun-Ching Shih |
| collection | DOAJ |
| description | Many elderly individuals frequently experience cataracts that interfere with vision. After cataract surgery, the left lens epithelial cell (LEC) exhibited fibrosis and posterior capsule opacification (PCO). Sometimes, there is a need for a second surgery; nevertheless, people try other methods, such as a good pharmacological agent, to treat PCO to reduce transforming growth factor-β2 (TGF-β2) amounts to avoid secondary surgery. The aim of the present study was to explore the potential anti-PCO activity of five 2,4-dihydro-3H-pyrazol-3-one (DHPO) derivatives in a TGF-β2-induced fibrogenesis SRA01/04 cell model. The 2-phenyl-5-propyl-DHPO (TSE; no. 2: TSE-2) compound showed the best activity of reduced expression levels of TGF-β2 among five derivatives and therefore was chosen to evaluate the anti-PCO activity and molecular mechanisms on the Sma and mad protein (SMAD) signaling pathway (including TGF-β2, SMADs, and the inhibition of nuclear translocation of SMADs), non-SMAD pathway proteins, including p-extracellular, regulated protein kinases (ERK) 1/2, or <i>p</i>-c-Jun N-terminal kinase (JUN) by Western blotting, PCR, or confocal immunofluorescence analyses. Following treatment with 10 μg/mL of the five compounds, the cells displayed great viability by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTT) assay. In this study, the result of lactate dehydrogenase (LDH) activity measurement did not affect the cytotoxicity of the five compounds. In TGF-β2-induced fibrogenesis in SRA01/04 cells, treatment with the TSE compound decreased the TGF-β2/SMAD signaling genes, including reduced mRNA or expression levels of TGF-β2, SMAD3, and SMAD4, leading to inhibition of TGF-β2-induced fibrogenesis. Our confocal immunofluorescence analyses demonstrated that TSE treatment displays a suppressive effect on SMAD2/3 or SMAD4 translocation to the nucleus. Furthermore, TSE treatment exhibits a reduction in the non-SMAD target gene expression levels of <i>p</i>- c-Jun N-terminal kinase (JUN), <i>p</i>- extracellular, regulated protein kinases (ERK)1/2, <i>p</i>- p38 mitogen-activated protein kinase (p38), <i>p</i>-phosphatidylinositol 3-kinase (PI3K), <i>p</i>-mammalian target of rapamycin complex (mTORC), <i>p</i>-Akt (Ser<sup>473</sup>), and <i>p</i>-Akt (Thr<sup>308</sup>). The overall effect of TSE is to reduce the expression levels of collagen I and fibrinogen (FN), thus contributing to antifibrotic effects in cell models mimicking PCO. Our findings reveal the benefits of TSE by regulating TGF-β/SMAD signaling and non-SMAD signaling-related gene proteins to display antifibrotic activity in cells for the possibility of preventing PCO after cataract surgery. |
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| spelling | doaj.art-ea8dfdb89ea44ba0b1a1b57f50ad76fd2023-11-23T23:34:20ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452022-10-0144105048506610.3390/cimb44100343Protective Effects of One 2,4-Dihydro-3H-Pyrazol-3-one Derivative against Posterior Capsular Opacification by Regulation of TGF-β2/SMADs and Non-SMAD Signaling, Collagen I, and Fibronectin ProteinsChun-Ching Shih0Chia-Yi Lee1Fung-Fuh Wong2Cheng-Hsiu Lin3Department of Nursing, College of Nursing, Central Taiwan University of Science and Technology, No.666 Buzih Road, Beitun District, Taichung City 40601, TaiwanInstitute of Medicine, Chung Shan Medical University, Taichung City 40201, TaiwanSchool of Pharmacy, China Medical University, No. 91 Hsueh-Shih Road, Taichung City 40402, TaiwanDepartment of Internal Medicine, Fengyuan Hospital, Ministry of Health and Welfare, Fengyuan District, Taichung City 42055, TaiwanMany elderly individuals frequently experience cataracts that interfere with vision. After cataract surgery, the left lens epithelial cell (LEC) exhibited fibrosis and posterior capsule opacification (PCO). Sometimes, there is a need for a second surgery; nevertheless, people try other methods, such as a good pharmacological agent, to treat PCO to reduce transforming growth factor-β2 (TGF-β2) amounts to avoid secondary surgery. The aim of the present study was to explore the potential anti-PCO activity of five 2,4-dihydro-3H-pyrazol-3-one (DHPO) derivatives in a TGF-β2-induced fibrogenesis SRA01/04 cell model. The 2-phenyl-5-propyl-DHPO (TSE; no. 2: TSE-2) compound showed the best activity of reduced expression levels of TGF-β2 among five derivatives and therefore was chosen to evaluate the anti-PCO activity and molecular mechanisms on the Sma and mad protein (SMAD) signaling pathway (including TGF-β2, SMADs, and the inhibition of nuclear translocation of SMADs), non-SMAD pathway proteins, including p-extracellular, regulated protein kinases (ERK) 1/2, or <i>p</i>-c-Jun N-terminal kinase (JUN) by Western blotting, PCR, or confocal immunofluorescence analyses. Following treatment with 10 μg/mL of the five compounds, the cells displayed great viability by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTT) assay. In this study, the result of lactate dehydrogenase (LDH) activity measurement did not affect the cytotoxicity of the five compounds. In TGF-β2-induced fibrogenesis in SRA01/04 cells, treatment with the TSE compound decreased the TGF-β2/SMAD signaling genes, including reduced mRNA or expression levels of TGF-β2, SMAD3, and SMAD4, leading to inhibition of TGF-β2-induced fibrogenesis. Our confocal immunofluorescence analyses demonstrated that TSE treatment displays a suppressive effect on SMAD2/3 or SMAD4 translocation to the nucleus. Furthermore, TSE treatment exhibits a reduction in the non-SMAD target gene expression levels of <i>p</i>- c-Jun N-terminal kinase (JUN), <i>p</i>- extracellular, regulated protein kinases (ERK)1/2, <i>p</i>- p38 mitogen-activated protein kinase (p38), <i>p</i>-phosphatidylinositol 3-kinase (PI3K), <i>p</i>-mammalian target of rapamycin complex (mTORC), <i>p</i>-Akt (Ser<sup>473</sup>), and <i>p</i>-Akt (Thr<sup>308</sup>). The overall effect of TSE is to reduce the expression levels of collagen I and fibrinogen (FN), thus contributing to antifibrotic effects in cell models mimicking PCO. Our findings reveal the benefits of TSE by regulating TGF-β/SMAD signaling and non-SMAD signaling-related gene proteins to display antifibrotic activity in cells for the possibility of preventing PCO after cataract surgery.https://www.mdpi.com/1467-3045/44/10/343cataractposterior capsule opacificationSma and mad protein (SMAD)s pathwaynon-SMADs pathwaytransforming growth factor type β2 (TGF-β2 |
| spellingShingle | Chun-Ching Shih Chia-Yi Lee Fung-Fuh Wong Cheng-Hsiu Lin Protective Effects of One 2,4-Dihydro-3H-Pyrazol-3-one Derivative against Posterior Capsular Opacification by Regulation of TGF-β2/SMADs and Non-SMAD Signaling, Collagen I, and Fibronectin Proteins Current Issues in Molecular Biology cataract posterior capsule opacification Sma and mad protein (SMAD)s pathway non-SMADs pathway transforming growth factor type β2 (TGF-β2 |
| title | Protective Effects of One 2,4-Dihydro-3H-Pyrazol-3-one Derivative against Posterior Capsular Opacification by Regulation of TGF-β2/SMADs and Non-SMAD Signaling, Collagen I, and Fibronectin Proteins |
| title_full | Protective Effects of One 2,4-Dihydro-3H-Pyrazol-3-one Derivative against Posterior Capsular Opacification by Regulation of TGF-β2/SMADs and Non-SMAD Signaling, Collagen I, and Fibronectin Proteins |
| title_fullStr | Protective Effects of One 2,4-Dihydro-3H-Pyrazol-3-one Derivative against Posterior Capsular Opacification by Regulation of TGF-β2/SMADs and Non-SMAD Signaling, Collagen I, and Fibronectin Proteins |
| title_full_unstemmed | Protective Effects of One 2,4-Dihydro-3H-Pyrazol-3-one Derivative against Posterior Capsular Opacification by Regulation of TGF-β2/SMADs and Non-SMAD Signaling, Collagen I, and Fibronectin Proteins |
| title_short | Protective Effects of One 2,4-Dihydro-3H-Pyrazol-3-one Derivative against Posterior Capsular Opacification by Regulation of TGF-β2/SMADs and Non-SMAD Signaling, Collagen I, and Fibronectin Proteins |
| title_sort | protective effects of one 2 4 dihydro 3h pyrazol 3 one derivative against posterior capsular opacification by regulation of tgf β2 smads and non smad signaling collagen i and fibronectin proteins |
| topic | cataract posterior capsule opacification Sma and mad protein (SMAD)s pathway non-SMADs pathway transforming growth factor type β2 (TGF-β2 |
| url | https://www.mdpi.com/1467-3045/44/10/343 |
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