Abstract:
OBJECTIVE To explore the effect of
Antrodia camphorata polysaccharide on the inflammatory response of dopaminergic neurons (DAN) cells induced by 6-OHDA through inhibiting ROS-NLRP3-caspase-1 pathway.
METHODS To isolate the DAN from midbrain. Parkinson cell model was constructed
in vitro by 6-OHDA. The cells were divided into normal group, model group, control group and experimental group. In the normal group, the DAN cells were cultured; in model group the DAN cells treated by 6-OHDA; in control group the DAN cells treated by ROS inhibitor NAC+6-OHDA; in experimental group, the DAN cells treated by 6-OHDA+
Antrodia camphorata polysaccharide. Cell viability was detected by CCK-8 assay, ROS level was detected by flow cytometry and immunofluorescence staining, cell apoptosis was detected by flow cytometry. Hoechst 33342 staining of living cells, the expression of NLRP3, caspase-1 and pro-caspase-1 in cells was detected by protein immunoblotting (Western-bolt), and the secretion level of IL-1β, IL-6 and IL-18 in the supernatant was detected by enzyme linked immunosorbent assay(Elisa).
RESULTS In the model group, 6-OHDA could induce the inflammatory response of DAN cells, the expression of ROS increased, the level of NLRP3-caspase-1 inflammatory corpuscle increased and the rate of apoptosis increased, which was significantly different from that of the normal group(
P<0.05). After the
Antrodia camphorata polysaccharide intervention, the level of ROS was down regulated, the level of NLRP3-caspase-1 inflammatory corpuscle was decreased, and the rate of apoptosis was down regulated, which was significantly different from that of the model group(
P<0.05).
CONCLUSION Antrodia camphorata polysaccharide can regulate the inflammatory response of DAN cells by inhibiting the regulation of 6-OHDA in ROS-NLRP3-caspase-1 pathway, which is one of the mechanisms of
Antrodia camphorata polysaccharide in the inflammatory response of Parkinson disease.