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25 February 2016 : Laboratory Research  

TRPC6 May Protect Renal Ischemia-Reperfusion Injury Through Inhibiting Necroptosis of Renal Tubular Epithelial Cells

BingBing ShenABCDEF, Yue HeABCDEF, Shan ZhouABCDEF, Hongwen ZhaoABCDEF, Mei MeiABCDEF, Xiongfei WuABCDEFG

DOI: 10.12659/MSM.897353

Med Sci Monit 2016; 22:633-641


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.

Keywords: Benzylamines - pharmacology, Blotting, Western, Cell Line, Cell Shape - drug effects, Down-Regulation - drug effects, Epithelial Cells - pathology, Fluorescent Antibody Technique, Imidazoles - pharmacology, Ischemia - pathology, Kidney Tubules - pathology, Necrosis, Oxygen, Protective Agents - metabolism, RNA, Small Interfering - metabolism, Reperfusion Injury - prevention & control, Sulfonamides - pharmacology, TRPC Cation Channels - metabolism, Up-Regulation - drug effects

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Med Sci Monit 2024; 30:e944927

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750