PURPOSE: To investigate the (i) effect(s) of cholinergics on the expression and secretion of transforming growth factor (TGF)-beta(2) in human retinal pigment epithelium (RPE) and (ii) mechanism of action of atropine in the treatment of myopia. MATERIALS AND METHODS: The RPE cell line, D407, was (i) treated with carbachol (10 microM), (ii) treated with atropine (10 nM-100 microM), or (iii) pre-treated with atropine (10 nM-100 microM) and then exposed to carbachol (10 microM). A no-treatment group served as control. Expression of TGF-beta(2), after stimulation at different time points (2, 4, 8, 16, 24, and 48 hr), was measured by RT-PCR and Western blot analysis. Secretion of TGF-beta(2) was determined by ELISA. RESULTS: Carbachol induced a time-dependent increase in the levels of TGF-beta(2) mRNA and protein in the cytoplasm (p < 0.001). ELISA assays showed a time-dependent increase in levels of TGF-beta(2) protein in the supernatant with carbachol treatment (p < 0.001). There was no change of TGF-beta(2) in the cytoplasm or supernatant with atropine alone (p > 0.05). The increased expression and secretion of TGF-beta(2) caused by carbachol were suppressed by atropine (in the range of 10 nM-100 microM) when compared to treatment with carbachol alone (p < 0.001). The stimulating effect of 10 microM carbachol was inhibited completely by 100 microM atropine. CONCLUSIONS: In RPE cells, atropine inhibits the expression and secretion of TGF-beta(2) by blocking the muscarinic acetylcholine receptor (mAChR), which may control the development of myopia.
PURPOSE: To investigate the (i) effect(s) of cholinergics on the expression and secretion of transforming growth factor (TGF)-beta(2) in human retinal pigment epithelium (RPE) and (ii) mechanism of action of atropine in the treatment of myopia. MATERIALS AND METHODS: The RPE cell line, D407, was (i) treated with carbachol (10 microM), (ii) treated with atropine (10 nM-100 microM), or (iii) pre-treated with atropine (10 nM-100 microM) and then exposed to carbachol (10 microM). A no-treatment group served as control. Expression of TGF-beta(2), after stimulation at different time points (2, 4, 8, 16, 24, and 48 hr), was measured by RT-PCR and Western blot analysis. Secretion of TGF-beta(2) was determined by ELISA. RESULTS:Carbachol induced a time-dependent increase in the levels of TGF-beta(2) mRNA and protein in the cytoplasm (p < 0.001). ELISA assays showed a time-dependent increase in levels of TGF-beta(2) protein in the supernatant with carbachol treatment (p < 0.001). There was no change of TGF-beta(2) in the cytoplasm or supernatant with atropine alone (p > 0.05). The increased expression and secretion of TGF-beta(2) caused by carbachol were suppressed by atropine (in the range of 10 nM-100 microM) when compared to treatment with carbachol alone (p < 0.001). The stimulating effect of 10 microM carbachol was inhibited completely by 100 microM atropine. CONCLUSIONS: In RPE cells, atropine inhibits the expression and secretion of TGF-beta(2) by blocking the muscarinic acetylcholine receptor (mAChR), which may control the development of myopia.
Authors: David Troilo; Earl L Smith; Debora L Nickla; Regan Ashby; Andrei V Tkatchenko; Lisa A Ostrin; Timothy J Gawne; Machelle T Pardue; Jody A Summers; Chea-Su Kee; Falk Schroedl; Siegfried Wahl; Lyndon Jones Journal: Invest Ophthalmol Vis Sci Date: 2019-02-28 Impact factor: 4.799
Authors: Pei-Chang Wu; Chia-Ling Tsai; Gabriel M Gordon; Shinwu Jeong; Tatsuo Itakura; Nitin Patel; Songtao Shi; M Elizabeth Fini Journal: Mol Vis Date: 2015-02-06 Impact factor: 2.367