Figure 8.

Primary mLECs isolated from C57BL/6 male or female mice revealed significantly reduced ROS level with increased expression of Bmal1 and Nrf2 and target antioxidants, Prdx6, Catalase and SOD1 in response to Metformin. Primary mLECs (5 × 10⁴) isolated from C57BL/6 male and female mice were cultured in 12-well culture plate and treated with Metformin for 24 h as shown in figure. (A) Increased ROS levels during aging was mitigated by Metformin treatment. ROS levels were measured using H2-DCF-DA dye methods. The histograms represent the mean ± S.D. from three independent experiments. Untreated vs. Metformin-treated; * p < 0.05; ** p < 0.001. (B–F) Total RNA isolated from Metformin-treated primary mLECs-derived from C57BL/6 mice of both sexes was subjected for mRNA analysis of Bmal1 and Nrf2, and their target antioxidant gene, Prdx6, Catalase and SOD1 with their specific primers as shown in Table 1. The data represent the mean ± S.D. from three independent experiments. Untreated vs. Metformin-treated; * p < 0.05; ** p < 0.001.