This study investigated the mechanisms underlying tubular apoptosis in diabetes by identifying proapoptotic genes that are differentially upregulated by reactive oxygen species in renal proximal tubular cells (RFTCs) in models of diabetes. Total RNAs isolated from renal proximal tubules (RPTs) of 20-week-old heterozygous db/m+, db/db, and db/db catalase (CAT)-transgenic (Tg) mice were used for DNA chip microarray analysis. Real-time quantitative PCR assays, immunohistochemistry, and mice rendered diabetic with streptozotocin were used to validate the proapoptotic gene expression in RPTs. Cultured rat RFTCs were used to confirm the apoptotic activity and regulation of proapoptotic gene expression. Additionally, studies in kidney tissues from patients with and without diabetes were used to confirm enhanced proapoptotic gene expression in RPTs. Bcl-2-modifying factor (Bmf) was differentially upregulated (P < 0.01) in RFTs of db/db mice compared with db/m+ and dbldb CAT-Tg mice and in RPTs of streptozotocin-induced diabetic mice in which insulin reversed this finding. In vitro, Bmf cDNA overexpression in rat RPTCs coimmunoprecipated with Bcl-2, enhanced caspase-3 activity, and promoted apoptosis. High glucose (25 mmol/L) induced Bmf mRNA expression in RPTCs, whereas rotenone, catalase, diphenylene iodinium, and apocynin decreased it. Knockdown of Bmf' with small interfering RNA reduced high glucose induced apoptosis in RFTCs. More important, enhanced Bmf expression was detected in RFTs of kidneys from patients with diabetes. These data demonstrate differential upregulation of Bmf in diabetic RPTs and suggest a potential role for Bmf in regulating RPTC apoptosis and tubular atrophy in diabetes. Diabetes 61:474-484, 2012
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Univ London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, EnglandUniv London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, England
Allen, DA
Harwood, SM
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Univ London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, EnglandUniv London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, England
Harwood, SM
Varagunam, M
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Univ London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, EnglandUniv London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, England
Varagunam, M
Raftery, MJ
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Univ London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, EnglandUniv London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, England
Raftery, MJ
Yaqoob, MM
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Univ London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, EnglandUniv London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, England
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Univ London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, EnglandUniv London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, England
Allen, DA
Harwood, SM
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Univ London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, EnglandUniv London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, England
Harwood, SM
Varagunam, M
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Univ London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, EnglandUniv London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, England
Varagunam, M
Raftery, MJ
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Univ London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, EnglandUniv London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, England
Raftery, MJ
Yaqoob, MM
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Univ London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, EnglandUniv London, Dept Expt Med & Nephrol, William Harvey Res Inst, St Bartholomews & Royal London Sch Med & Dent, London EC1A 7BE, England