title: TRPC6 May Protect Renal Ischemia-Reperfusion Injury Through Inhibiting Necroptosis of Renal Tubular Epithelial Cells.
abstract: BACKGROUND The aim of this study was to explore the potential role of TRPC6 in the pathophysiology of HK-2 cell injury following ischemia reperfusion (I/R). MATERIAL AND METHODS TRPC6 expression was analyzed by immunofluorescence staining. siRNA was transfected to knockout of TRPC6 in HK-2 cells, and in vitro I/R was then induced. Cell apoptosis and necrosis were determined by Annexin V-FITC/PI staining. Necroptosis was determined by necrostatin-1 and expressions of necroptosis-related proteins were evaluated. OAG, SKF96365, or KN-93 was further used to interfere with TRPC6 expression. RESULTS Cytoplasmic TRPC6 expression was demonstrated. I/R induced TRPC6 expression in normal or NC siRNA-transfected cells but not in TRPC6 siRNA-knockout ones. There was a progressive increase in apoptotic and necrotic cells with increasing reoxygenation time in all 3 groups, while necrosis in TRPC6 siRNA-transfected cells was comparatively higher than that of the other 2 groups (p<0.05). Expressions of necroptosis-related proteins were interfered with following I/R and these effects were enhanced by TRPC6 siRNA. Application of OAG, SKF96365, or KN93 further affected necroptosis following I/R. CONCLUSIONS This study described the expression and functional relevance of TRPC6 in the pathophysiology of HK-2 cell following I/R. Our results regarding the ability of TRPC6 to specifically interrupt necroptosis may shed new light on its role in prevention and control of ischemic kidney injury.
authors.affiliation: Department of Kidney, Southwest Hospital, Third Military Medical University, Chongqing, China (mainland).
authors.forename: BingBing
authors.initials: B
authors.lastname: Shen
authors.affiliation: Department of Kidney, Southwest Hospital, Third Military Medical University, Chongqing, China (mainland).
authors.forename: Yue
authors.initials: Y
authors.lastname: He
authors.affiliation: Department of Kidney, Southwest Hospital, Third Military Medical University, Chongqing, China (mainland).
authors.forename: Shan
authors.initials: S
authors.lastname: Zhou
authors.affiliation: Department of Kidney, Southwest Hospital, Third Military Medical University, Chongqing, China (mainland).
authors.forename: Hongwen
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Department of Kidney, Southwest Hospital, Third Military Medical University, Chongqing, China (mainland).
authors.forename: Mei
authors.initials: M
authors.lastname: Mei
authors.affiliation: Department of Kidney, Southwest Hospital, Third Military Medical University, Chongqing, China (mainland).
authors.forename: Xiongfei
authors.initials: X
authors.lastname: Wu
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001596:Benzylamines / Q000494:pharmacology; D015153:Blotting, Western; D002460:Cell Line; D048430:Cell Shape / Q000187:drug effects; D015536:Down-Regulation / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007511:Ischemia / Q000473:pathology*; D007684:Kidney Tubules / Q000098:blood supply* / Q000473:pathology*; D009336:Necrosis; D010100:Oxygen; D020011:Protective Agents / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D015427:Reperfusion Injury / Q000473:pathology / Q000517:prevention & control*; D013449:Sulfonamides / Q000494:pharmacology; D050052:TRPC Cation Channels / Q000378:metabolism*; D000074287:TRPC6 Cation Channel; D015854:Up-Regulation / Q000187:drug effects; D000096926:Benzenesulfonamides
chemical_list: D001596:Benzylamines; D007093:Imidazoles; D010100:Oxygen; D020011:Protective Agents; D034741:RNA, Small Interfering; D013449:Sulfonamides; D050052:TRPC Cation Channels; D000074287:TRPC6 Cation Channel; C117032:TRPC6 protein, human; C072105:KN 93; D000096926:Benzenesulfonamides; C063159:1-(2-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenylethyl)-1H-imidazole