Title : Korean medicinal herbs inhibit MPP+-induced mitochondrial dysfunction in SH-SY5Y human neuroblastoma cells
Abstract:
These days, interplay between brain and gut is important role in brain related disorders. Immune system and tight junctions in gut are thought that there is a correlation with Central nerve system and brain. Two kinds of medicinal herbs, Rumex japonicus Houtt. (RJ) and Atractylodes macrocephala Koidz. (AM), has been used to treat various digestive disease and inflammation in Korea. Also, RJ and AM has anti-oxidative and anti-inflammatory effect. Therefore, RJ and AM are expected to improve not only digestive disease but also brain disorders such as Parkinson’s disease (PD). In the present study, we evaluated the protective effect of RJ and AM against neurotoxin 1-methyl-4-phenylpyridinium ion (MPP+)-induced mitochondrial dysfunction in SH-SY5Y human neuroblastoma cells, an in vitro PD model. SH-SY5Y cells were incubated with SF for 24 h, after which they were treated with MPP+. MPP+-induced cytotoxicity and apoptosis were confirmed by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling assay. Immunofluorescent staining is conducted to confirm the PD related factors (Parkin, PINK1 and DJ-1), reactive oxidative species (ROS), and permeability in mitochondria. Also, ATP assay and western blot was performed to evaluate mitochondrial function-related factors (Parkin, PINK1 and DJ-1) and apoptosis factors (caspase-3 and cytochrome c, Bcl-2 and Bax,). Methanol extracts (0.01 mg/mL of RJ; 0.01 mg/mL of AM) suppressed MPP+-induced cytotoxicity and apoptosis. Also, RJ and AM effectively inhibited collapse of mitochondrial membrane potential and increase of ROS. In the result of western blot, RJ and AM regulated the expressions of apoptosis (caspase-3 and cytochrome c, Bcl-2 and Bax,) and PD (Parkin, PINK1 and DJ-1) related proteins. The SH-SY5Y cells treated with RJ or AM showed higher ATP activity than MPP+ only treated group. Therefore, RJ and AM effectively suppressed MPP+-induced cytotoxicity, apoptosis and mitochondrial dysfunction, and loss or mutation of mitochondria-related PD markers.
