Fig. 5.

Dose- and time-dependent impairments of mitochondrial biogenesis in rat cardiomyocytes induced by ADMA and OA. Rat cardiomyocytes were incubated with various dose of high glucose (15 ~ 30 mM) and asymmetric dimethylarginine (ADMA, 3 ~ 100 μM) or oleic acid (OA, 10 ~ 100 μM) and for different time (12 ~ 72 h) as indications. The content of ADMA was measured by enzyme linked immunosorbent assay (ELISA) and normalized to the protein content of the supernatants of cell lysis. The ratio of the copy number of mitochondrial gene COX Ι to nuclear gene β-actin was used to evaluate the mitochondrial biogenesis. Panel a & b show the dose-response and time course of high glucose and OA on ADMA elaboration; Panel c & d represent the dose-response and time course of ADMA on mitochondrial biogenesis; Panel e displays the dose-response of OA, and panel f displays effects of ADMA combination with high glucose or OA or even both on mitochondrial biogenesis in cardiomyocytes. Graphs in panel c, d, e & f represent the quantifications of COX Ι vs β-actin from 3 independent experiments, respectively. Date are expressed as mean ± S.E.M., n = 3, ^*P < 0.05, ^**P < 0.01 vs control or zero group