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Research Paper Volume 12, Issue 6 pp 4836—4865

Nrf2 deficiency aggravates PM2.5-induced cardiomyopathy by enhancing oxidative stress, fibrosis and inflammation via RIPK3-regulated mitochondrial disorder

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Figure 8. Nrf2 modulates RIPK3 expression to regulate mitochondrial disorder in PM2.5-exposed cardiomyocytes. (A) Cardiomyocytes were treated with the indicated concentrations of GSK872 (0, 1.25, 2.5 and 5 μM) for 2 h, followed by western blot analysis of RIPK3. n = 6 in each group. (B–G) Cardiomyocytes isolated from Nrf2+/+ or Nrf2-/- mice were pre-treated with GSK872 (15 μM) for 2 h, or transfected with si-RIPK3 for 24 h. Then, all cells were exposed to PM2.5 (100 μg/ml) for another 24 h. After treatments above, all cells were collected for further studies. (B) Mitochondrial potential results by JC-1 analysis. n = 8 in each group. (C) Intracellular ATP levels were measured. n = 8 in each group. (D) Results of mPTP opening in cells. n = 8 in each group. (E) RT-qPCR analysis of Fis1, Drp1, Mid51, Mid49, MFN1, MFN2 and Opa1 in cells. n = 6 in each group. (F, G) Western blot analysis of NDUFB8, SDHB, UQCRC1 and MTCO1 in the mitochondrial fractions. n = 6 in each group. Data were expressed as the mean ± SEM. *P < 0.05 and **P < 0.01.