Fig. 7. The effect of GS on mitochondrial function was
abrogated by NAM, a SIRT1 inhibitor. (A-B) Basal OCR and MRC in
H9c2 cells were measured, after the treatment by GS and/or NAM for 48 h.(C) H9c2 cells were treated with GS and/or NAM for 48 h to
measure ATP level by a bioluminescence detection kit. (D) The
expressions of CV-ATP5A, CIII-UQCRC2, CIV-MTCO1, CII-SDHB, and CI-NDUFB8
in H9c2 cells treated with GS and/or NAM for 48 h were detected by
Western blot. (E) Relative levels of five mitochondrial
complexes from (D) are shown, after the quantification by ImageJ.(F) The effect of GS combined with NAM on the
NAD+ level was analyzed by a NAD/NADH-Glo assay kit.(G-H) Total proteins of H9c2 cells treated with GS and/or NAM
for 48 h were prepared and followed by Western blot analysis using
antibodies against SIRT1, PGC1α, Nrf1, Nrf1, and β-Actin. Relative
expression of these proteins was quantified with ImageJ and shown on the
right. β-Actin is loading control.
Ctrl: control group, OCR: oxygen consumption rate, MRC: maximal
respiration capacity. *P < 0.05 and***P < 0.001 versus Ctrl
group; #P < 0.05 and##P < 0.01 versus GS+NAM
group.