Fig. 1. GS enhanced mitochondrial respiration capacity and ATP
production in cardiomyocytes and neurons. (A-B) Basal oxygen
consumption rate (OCR) in H9c2, NGF-induced PC12, neuron, HUVECs and
BMSCs treated with different concentrations of total ginsenosides (GS)
for 48 h was measured using a
MitoXpress® Xtra probe normalized to cell number. (C)OCR was analyzed at basal
condition and sequential injections of oligomycin (1 µM), FCCP (5 μM),
rotenone (Rot, 1 μM) and antimycin A (AA, 1 μM) in control and
GS-pretreated H9c2 cells by the MitoStress Test Kit. After the
normalization to cell number, basal OCR, ATP production, maximal
respiratory capacity (MRC), and spare respiratory capacity (SRC) were
calculated and shown. (D) Similar to the panel C, basal OCR,
ATP production, MRC, and SRC were analyzed in NGF-induced PC12 cells,
after GS pretreatment for 48 h. (E) After pretreatment with GS
at 2.5, 5, or 10 μg/mL for 48 h, ATP levels in H9c2, PC12, and neurons
were measured by the Bioluminescence Detection Kit. (F)The intensities of ATP and ADP in
control and GS-treated H9c2 cells were analyzed by the LC-MS/MS method
based on MRM detection. The ratio of ATP and ADP was calculated and
shown. (G-H) Basal OCR and ATP content in H9c2 cells treated
with different concentrations (2.5, 5, or 10 μg/mL) of GS and incubated
for 3, 6, 12, 24, or 48 h were measured as described as the methods
above. Ctrl: control group. *P < 0.05, **P< 0.01, and ***P < 0.001 versus Ctrl
group.