PMID- 32116675 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20200928 IS - 1663-9812 (Print) IS - 1663-9812 (Electronic) IS - 1663-9812 (Linking) VI - 10 DP - 2019 TI - High Glucose-Induced TRPC6 Channel Activation Decreases Glutamate Uptake in Rat Retinal Muller Cells. PG - 1668 LID - 10.3389/fphar.2019.01668 [doi] LID - 1668 AB - High glucose (HG) increases the production of reactive oxygen species (ROS), leading to decreased glutamate uptake in Muller cells. The transient receptor potential cation channel 6 (TRPC6) channel, an oxidative stress-sensitive Ca(2+)-permeable cationic channel, is readily detected in Muller cells and highly expressed under HG conditions. Yet, the effect of high glucose-induced TRPC6 channel activation in Muller cells is poorly understood. We hypothesized that TRPC6 channel activation mediates high glucose-induced decreases in Muller cell glutamate uptake. We found RNA interference (RNAi) of the TRPC6 channel abolished HG-induced decreases in glutamate uptake and cell death. HG also decreased the expression of the glutamate-aspartate transporter (GLAST), which is the most important transporter involved in glutamate uptake. The mRNA level of ciliary neurotrophic factor (CNTF) in rMC-1 cells and the release of CNTF in the culture media was decreased, but the mRNA levels of IL-6 and vascular endothelial growth factor (VEGF) were increased under HG conditions. After RNAi silencing in rMC-1 cells, the mRNA levels of CNTF increased, but IL-6 and VEGF levels decreased. Furthermore, TRPC6 knockdown (KD) decreased expression of glial fibrillary acidic protein (GFAP) and increased expression of Kir4.1, pointing to inhibition of HG-induced gliosis in rMC-1 cells. ROS and intracellular Ca(2+) levels decreased after TRPC6 knockdown. Exposure to Hyp9 (10 muM), a highly selective TRPC6 channel agonist, can aggravate HG-induced pathological changes. Collectively, our results suggest TRPC6 channel activation is involved in HG-induced decreases in glutamate uptake in rMC-1 cells. These findings provide novel insights into the role of TRPC6 in HG-induced retinal neurovasculopathy and suggest TRPC6 is a promising target for drug development for diabetic retinopathy (DR). CI - Copyright (c) 2020 Ma, Zhao, Zhang, Sun, Fan and Zheng. FAU - Ma, Mingming AU - Ma M AD - Department of Ophthalmology, Shanghai General Hospital, Shanghai, China. AD - National Clinical Research Center for Eye Diseases, Shanghai, China. AD - Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, China. FAU - Zhao, Shuzhi AU - Zhao S AD - Department of Ophthalmology, Shanghai General Hospital, Shanghai, China. AD - National Clinical Research Center for Eye Diseases, Shanghai, China. AD - Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, China. FAU - Zhang, Jian AU - Zhang J AD - Department of Ophthalmology, Shanghai General Hospital, Shanghai, China. AD - National Clinical Research Center for Eye Diseases, Shanghai, China. AD - Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, China. FAU - Sun, Tao AU - Sun T AD - Department of Ophthalmology, Shanghai General Hospital, Shanghai, China. AD - National Clinical Research Center for Eye Diseases, Shanghai, China. AD - Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, China. FAU - Fan, Ying AU - Fan Y AD - Department of Ophthalmology, Shanghai General Hospital, Shanghai, China. AD - National Clinical Research Center for Eye Diseases, Shanghai, China. AD - Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, China. FAU - Zheng, Zhi AU - Zheng Z AD - Department of Ophthalmology, Shanghai General Hospital, Shanghai, China. AD - National Clinical Research Center for Eye Diseases, Shanghai, China. AD - Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, China. AD - Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, China. LA - eng PT - Journal Article DEP - 20200214 PL - Switzerland TA - Front Pharmacol JT - Frontiers in pharmacology JID - 101548923 PMC - PMC7033573 OTO - NOTNLM OT - Muller cells OT - TRPC6 channel OT - diabetic retinopathy OT - glutamate uptake OT - reactive oxygen species EDAT- 2020/03/03 06:00 MHDA- 2020/03/03 06:01 PMCR- 2020/02/14 CRDT- 2020/03/03 06:00 PHST- 2019/09/16 00:00 [received] PHST- 2019/12/20 00:00 [accepted] PHST- 2020/03/03 06:00 [entrez] PHST- 2020/03/03 06:00 [pubmed] PHST- 2020/03/03 06:01 [medline] PHST- 2020/02/14 00:00 [pmc-release] AID - 10.3389/fphar.2019.01668 [doi] PST - epublish SO - Front Pharmacol. 2020 Feb 14;10:1668. doi: 10.3389/fphar.2019.01668. eCollection 2019.