Triamcinolone acetonide modulates TGF‑β2‑induced angiogenic and tissue‑remodeling effects in cultured human retinal pigment epithelial cells

Hsiao,Chih-Cheng; Chang,Yo-Chen; Hsiao,Yu-Ting; Chen,Po-Han; Hsieh,Ming-Chu; Wu,Wen-Chuan; Kao,Ying-Hsien

doi:10.3892/mmr.2021.12442

Triamcinolone acetonide modulates TGF‑β2‑induced angiogenic and tissue‑remodeling effects in cultured human retinal pigment epithelial cells

Authors:
- Chih-Cheng Hsiao
- Yo-Chen Chang
- Yu-Ting Hsiao
- Po-Han Chen
- Ming-Chu Hsieh
- Wen-Chuan Wu
- Ying-Hsien Kao
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Affiliations: Department of Pediatrics, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan, R.O.C., Department of Ophthalmology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan, R.O.C., Department of Ophthalmology, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung 83301, Taiwan, R.O.C., Department of Medical Research, E‑Da Hospital, Kaohsiung 82445, Taiwan, R.O.C., Department of Ophthalmology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung 80761, Taiwan, R.O.C., Department of Ophthalmology, China Medical University Hospital, Taichung 404332, Taiwan, R.O.C.
Published online on: September 14, 2021 https://doi.org/10.3892/mmr.2021.12442
Article Number: 802
Copyright: © Hsiao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Transforming growth factor‑β2 (TGF‑β2) has been implicated in the pathogenesis of proliferative vitreoretinopathy (PVR) and proliferative diabetic retinopathy (PDR), due to its ability to stimulate the overproduction of pro‑angiogenic factors, such as vascular endothelial growth factor (VEGF), and remodeling of the extracellular matrix (ECM). Although intravitreal triamcinolone acetonide (TA) is clinically useful in the treatment of PVR and PDR, its molecular mechanism has yet to be fully elucidated. The present study investigated whether TA treatment altered TGF‑β2‑driven biological effects on the behavior of cultured human retinal pigment epithelial (RPE) cells, in order to determine which signaling pathway may be essential for the pharmacological action of TA. The R‑50 human RPE cell line was treated with TA in the presence of TGF‑β2, followed by analyses of cell viability and contraction using cell viability and collagen gel contraction assays. VEGF mRNA expression and protein production were measured using reverse transcription‑quantitative PCR and ELISA, respectively. The phosphorylation status of signaling mediators and the protein expression of type I collagen (COL1A1), α‑smooth muscle actin (α‑SMA), and ECM‑remodeling enzymes, including MMP‑2 and MMP‑9, were analyzed using western blotting. The gelatinolytic activity of MMPs was detected using gelatin zymography. TA treatment exhibited no prominent cytotoxicity but markedly antagonized TGF‑β2‑induced cytostatic effects on RPE cell viability and TGF‑β2‑enhanced contractility in collagen gels. In the context of TGF‑β2‑related signaling, TA significantly attenuated TGF‑β2‑elicited Smad2, extracellular‑regulated kinase (ERK)1/2 and p38 mitogen‑activated protein kinase (MAPK) phosphorylation. Moreover, TA markedly mitigated TGF‑β2‑induced VEGF upregulation through ablation of p38 signaling activity. TA also partially attenuated TGF‑β2‑elicted expression of COL1A1, α‑SMA, MMP‑2, and MMP‑9, but only suppressed TGF‑β2‑induced MMP‑9 gelatinolytic activity. Mechanistically, the MEK/ERK signaling pathway may have a critical role in the TGF‑β2‑induced upregulation of COL1A1, α‑SMA and MMP‑9. In conclusion, TA may be considered a useful therapeutic agent for treating TGF‑β2‑associated intraocular angiogenesis and tissue remodeling, the underlying mechanism of which may involve the ERK and p38 MAPK signaling pathways.

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