Detecting Cellular Hypoxia in HRPTECs and Rats

Cellular hypoxia was detected by adding pimonidazole hydrochloride (200 mmol/L hypoxyprobe-1; Hydroxyprobe, Burlington, MA), which binds to cells or tissues with pO₂ levels <10 mmHg, to HRPTECs that were treated with 1 mmol/L metformin or 1 mmol/L AICAR and exposed to hypoxia (1% O₂) for 4 h. To detect hypoxic conditions in each group of rats, pimonidazole (60 mg/kg) was injected intraperitoneally 1 h before they were killed. Staining was performed according to the manufacturer’s instructions.

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Takiyama Y., Harumi T., Watanabe J., Fujita Y., Honjo J., Shimizu N., Makino Y, & Haneda M. (2011). Tubular Injury in a Rat Model of Type 2 Diabetes Is Prevented by Metformin: A Possible Role of HIF-1α Expression and Oxygen Metabolism. Diabetes, 60(3), 981-992.

Publication 2011

Aicar Cells Cellular hypoxia Hypoxic Hypoxyprobe 1 Metformin Pimonidazole Rats Tissues

Corresponding Organization : Asahikawa Medical University

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Variable analysis

independent variables

Treatment with 1 mmol/L metformin
Treatment with 1 mmol/L AICAR
Exposure to hypoxia (1% O2) for 4 h

dependent variables

Cellular hypoxia detected by pimonidazole hydrochloride binding

control variables

HRPTECs (human renal proximal tubular epithelial cells)

controls

Positive control: Pimonidazole (60 mg/kg) injected intraperitoneally 1 h before euthanasia to detect hypoxic conditions in each group of rats

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