Figure 9.
Metformin enhanced the expression of Bmal1 and Nrf2, and their target antioxidant genes, phase II enzymes in primary embryonic human lens epithelial cells (Emb-hLECs). (A–E) Metformin amplified expression of Bmal1, Nrf2 and its target genes, Prdx6, Catalase and SOD1 mRNA in Emb-hLECs. Emb-hLECs (2 × 105) were cultured in 35 mm culture plate and treated with Metformin as indicated. Total RNA was isolated and processed for RT-qPCR analyses using primers specific to above-mentioned molecules. The histograms represent the mean ± S.D. from three independent experiments. Untreated vs. Metformin-treated; ** p < 0.001. (F) Metformin enhanced the expression of Bmal1, Nrf2 and its antioxidant target protein, such as Prdx6 in Emb-hLECs. Emb-hLECs (2 × 105) were cultured in 35 mm culture plate and treated with Metformin. Total protein was isolated and subjected to Western analyses. Tubulin was used as internal control. Protein bands were quantified using a densitometer, and levels were normalized to corresponding tubulin levels and presented as histograms in the right side of the protein bands. Untreated vs. Metformin-treated; ** p < 0.001. (G–J) Metformin treatment increased the activities of aiPLA2, GSH Peroxidase, Catalase and SOD in Emb-hLECs. Emb-hLECs (2 × 105) were cultured in 35 mm culture plate and treated with Metformin as shown in Figure. Total protein was isolated and enzymatic activities of antioxidants were estimated and presented as indicated. The data represent the mean ± S.D. from three independent experiments. Untreated vs. Metformin-treated; * p < 0.05; ** p < 0.001.