PMID- 37723490 OWN - NLM STAT- MEDLINE DCOM- 20230928 LR - 20231121 IS - 1478-811X (Electronic) IS - 1478-811X (Linking) VI - 21 IP - 1 DP - 2023 Sep 18 TI - Arginase-1 promotes lens epithelial-to-mesenchymal transition in different models of anterior subcapsular cataract. PG - 236 LID - 10.1186/s12964-023-01210-4 [doi] LID - 236 AB - BACKGROUND: Arginase-1 (ARG1) promotes collagen synthesis and cell proliferation. ARG1 is highly expressed in various tumour cells. The mechanisms of ARG1 in epithelial-to-mesenchymal transition (EMT)-associated cataracts were studied herein. METHODS: C57BL/6 mice, a human lens epithelial cell line (HLEC-SRA01/04), and human lens capsule samples were used in this study. The right lens anterior capsule of the mouse eye was punctured through the central cornea with a 26-gauge hypodermic needle. Human lens epithelial cells (HLECs) were transfected with ARG1-targeted (siARG1) or negative control siRNA (siNC). For gene overexpression, HLECs were transfected with a plasmid bearing the ARG1 coding sequence or an empty vector. Medium containing 0.2% serum with or without transforming growth factor beta-2 (TGF-beta2) was added for 6 or 24 h to detect mRNA or protein, respectively. The expression of related genes was measured by quantitative real-time polymerase chain reaction (RT-qPCR), western blotting, and immunohistochemical staining. Transwell assays and wound healing assays were used to determine cell migration. Cell proliferation, superoxide levels, nitric oxide (NO) levels, and arginase activity were estimated using Cell Counting Kit-8 assays, a superoxide assay kit, an NO assay kit, and an arginase activity kit. RESULTS: ARG1, alpha-smooth muscle actin (alpha-SMA), fibronectin, and Ki67 expression increased after lens capsular injury, while zonula occludens-1 (ZO-1) expression decreased. Fibronectin and collagen type I alpha1 chain (collagen 1A1) expression increased, and cell migration increased significantly in ARG1-overexpressing HLECs compared with those transfected with an empty vector after TGF-beta2 treatment. These effects were reversed by ARG1 knockdown. The arginase-related pathway plays an important role in EMT. mRNAs of enzymes of the arginase-related pathway were highly expressed after ARG1 overexpression. ARG1 knockdown suppressed these expression changes. Numidargistat (CB-1158) dihydrochloride (CB-1158), an ARG1 inhibitor, suppressed TGF-beta2-induced anterior subcapsular cataract (ASC) by reducing the proliferation of lens epithelial cells (LECs) and decreasing fibronectin, alpha-SMA, collagen 1A1, and vimentin expression. Compared with that in nonanterior subcapsular cataract (non-ASC) patients, the expression of ARG1, collagen 1A1, vimentin, fibronectin, and Ki67 was markedly increased in ASC patients. CONCLUSIONS: ARG1 can regulate EMT in EMT-associated cataracts. Based on the pathogenesis of ASC, these findings are expected to provide new therapeutic strategies for patients. CI - (c) 2023. BioMed Central Ltd., part of Springer Nature. FAU - Li, Qingyu AU - Li Q AD - Department of Cataract, Tianjin Eye Hospital, Tianjin, China. AD - Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin, China. FAU - Wang, Yuchuan AU - Wang Y AD - Department of Cataract, Tianjin Eye Hospital, Tianjin, China. AD - Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin, China. FAU - Shi, Luoluo AU - Shi L AD - Department of Cataract, Tianjin Eye Hospital, Tianjin, China. AD - Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin, China. FAU - Wang, Qing AU - Wang Q AD - Clinical College of Ophthalmology, Tianjin Medical University, Tianjin, China. AD - Heze Medical College, Heze, Shandong, China. FAU - Yang, Guang AU - Yang G AD - School of Microelectronics, Tianjin University, Tianjin, China. FAU - Deng, Lin AU - Deng L AD - Department of Cataract, Tianjin Eye Hospital, Tianjin, China. AD - Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin, China. FAU - Tian, Ye AU - Tian Y AD - Department of Cataract, Tianjin Eye Hospital, Tianjin, China. AD - Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin, China. FAU - Hua, Xia AU - Hua X AD - Tianjin Aier Eye Hospital, Tianjin University, Tianjin, China. cathayhuaxia@163.com. FAU - Yuan, Xiaoyong AU - Yuan X AD - Department of Cataract, Tianjin Eye Hospital, Tianjin, China. yuanxy_cn@hotmail.com. AD - Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin, China. yuanxy_cn@hotmail.com. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20230918 PL - England TA - Cell Commun Signal JT - Cell communication and signaling : CCS JID - 101170464 RN - EC 3.5.3.1 (Arginase) RN - 0 (Ki-67 Antigen) RN - 11062-77-4 (Superoxides) RN - 0 (Transforming Growth Factor beta2) RN - 0 (Vimentin) RN - EC 3.5.3.1 (ARG1 protein, human) RN - EC 3.5.3.1 (Arg1 protein, mouse) SB - IM MH - Animals MH - Humans MH - Mice MH - *Arginase MH - *Cataract MH - Ki-67 Antigen MH - Mice, Inbred C57BL MH - Superoxides MH - Transforming Growth Factor beta2 MH - Vimentin MH - *Epithelial-Mesenchymal Transition PMC - PMC10506332 OAB - Fibrotic cataracts can be classified as anterior subcapsular cataract or posterior capsular opacification depending on where fibrosis occurs. The mechanism of fibrotic cataracts is not fully understood. Fibrotic opacities induced by trauma, inflammation, or radiation can accumulate underneath the anterior lens capsule, causing anterior subcapsular cataract. Posterior capsular opacification is one of the most common complications of phacoemulsification with intraocular lens implantation, with a high incidence in young patients. We show for the first time that ARG1 can regulate EMT in fibrotic cataracts. TGF-beta2 is the main cause of fibrosis in LECs. The expression of ARG1 and fibronectin in LECs increased after TGF-beta2 treatment or mouse lens capsular injury. We investigated the specific molecular mechanisms by which ARG1 regulates EMT in fibrotic cataracts. The mRNA expression of enzymes of the arginase-related pathway was decreased due to knockdown of ARG1 expression in HLECs. These effects were reversed by ARG1 overexpression. Additionally, knockdown of ARG1 decreased collagen 1A1, fibronectin, and vimentin expression; superoxide levels; and cell migration and increased NO levels. These effects were reversed by ARG1 overexpression. Pharmacological blockade of the ARG1 pathway with CB-1158 reduced the proliferation of LECs and decreased fibronectin, alpha-SMA, collagen 1A1, and vimentin expression in mouse lenses. We believe that ARG1 promotes the production of collagen 1A1 by directly activating the arginase pathway and leads to lens fibrosis by reducing NO production and increasing superoxide levels, providing a new mechanism for the prevention and treatment of fibrotic cataracts. Video Abstract. OABL- eng OTO - NOTNLM OT - ARG1 OT - Anterior subcapsular cataract OT - Arginase-related pathway OT - CB-1158 OT - Posterior capsular opacification COIS- The authors declare that they have no competing interests. EDAT- 2023/09/19 00:43 MHDA- 2023/09/20 06:42 PMCR- 2023/09/18 CRDT- 2023/09/18 23:47 PHST- 2022/12/23 00:00 [received] PHST- 2023/06/30 00:00 [accepted] PHST- 2023/09/20 06:42 [medline] PHST- 2023/09/19 00:43 [pubmed] PHST- 2023/09/18 23:47 [entrez] PHST- 2023/09/18 00:00 [pmc-release] AID - 10.1186/s12964-023-01210-4 [pii] AID - 1210 [pii] AID - 10.1186/s12964-023-01210-4 [doi] PST - epublish SO - Cell Commun Signal. 2023 Sep 18;21(1):236. doi: 10.1186/s12964-023-01210-4.