Figure 7.

(A–D) Metformin augmented the expression of Bmal1, Nrf2 and target antioxidant gene, Prdx6 in mouse LECs. mLECs (5 × 10⁵) were cultured in 60 mm culture plate and treated with Metformin for 24 h. Total RNA (A–C) and protein (D) were extracted and submitted to RT-qPCR and Western blot analysis with probes specific to corresponding molecules indicated. Bmal1, Nrf2 and Prdx6 transcripts and protein expression were significantly increased in response to Metformin. The data represent the mean ± S.D. from three independent experiments. Untreated vs. Metformin-treated; ** p < 0.001. (E–H) Metformin amplified Bmal1/Nrf2 target antioxidants enzymatic activities. Equal amounts of protein isolated from mLECs were processed to analyze the enzymatic activities using a commercially available kit. Metformin treatment significantly increased cellular antioxidant enzymatic activities examined; PLA₂ €, GSH peroxidase (F), Catalase (G) and SOD (H). The data represent the mean ± S.D. from three independent experiments. Untreated vs. Metformin-treated; * p < 0.05; ** p < 0.001.