ABSTRACT
Objective:
To investigate the protective effect and molecular mechanism of Angelicae Sinensis Radix-Chuanxiong Rhizoma medicated
serum (ASRCRS) against
oxidative damage of
PC12 cells induced by H<sub>2</sub>O<sub>2</sub>.
Method:
Oxidative damage of
PC12 cells was induced by H<sub>2</sub>O<sub>2</sub><italic>
in vitro</italic>, and intervention was performed in the low-, medium-, and high-
dose ASRCRS groups with a final volume fraction of 15%. The
cell viability was determined by methyl thiazolyl tetrazolium (MTT) assay.
Cell morphology was observed by an inverted
fluorescence microscope. The content of
lactate dehydrogenase (LDH) and
malondialdehyde (MDA), the activity of
superoxide dismutase (SOD), and the distribution of
reactive oxygen species (ROS) in the
cell supernatant were detected by the kits.
Cell apoptosis was detected by
Annexin V-
FITC/PI double
staining. The
protein expression levels of nuclear factor E<sub>2</sub>-related factor 2 (Nrf2), Kelch-like
epichlorohydrin associated
protein-1 (Keap1),
heme oxygenase-1 (HO-1), and SOD1 were detected by
Western blot.
Result:
Oxidative damage was induced by 300 μmol·L<sup>-1</sup> H<sub>2</sub>O<sub>2</sub> for 24 hours. Compared with the normal group, the model group showed abnormal
cell morphology, reduced
cell viability (<italic>P</italic><0.01), increased LDH and MDA (<italic>P</italic><0.01), blunted SOD activity, elevated intracellular distribution of ROS, down-regulated
protein expression of Nrf2, HO-1, and SOD1 (<italic>P</italic><0.05, <italic>P</italic><0.05), and up-regulated
protein expression of Keap1 (<italic>P</italic><0.01). Compared with the model group, ASRCRS groups displayed improved
cell morphology, increased
cell viability, inhibited
cell apoptosis, potentiated SOD activity (<italic>P</italic><0.01), suppressed release of LDH (<italic>P</italic><0.01) and generation of ROS, decreased content of MDA (<italic>P</italic><0.01), up-regulated
protein expression of Nrf2, HO-1 and SOD1 (<italic>P</italic><0.05, <italic>P</italic><0.01), and down-regulated
protein expression of Keap1 (<italic>P</italic><0.01).
Conclusion:
ASRCRS could protect
PC12 cells from
oxidative damage induced by H<sub>2</sub>O<sub>2</sub> by up-regulating the expression of Nrf2 to activate the Nrf2/
antioxidant response element (ARE) signaling pathway, enhancing the
ability to resist
oxidative damage, and inhibiting
cell apoptosis.