Figure 1 Therapeutic effect of Atractylodis Rhizoma Alba extracts in a Parkinson’s disease murine model A: schematic diagram of drug treatment, behavioral test, and tissue preparation; B: MPTP treatment decreases the total distance in the center of the OFT. ARE increases the distance in the center area of OFT; C: motor skill learning of the MPTP-induced mice in the rotarod test provides evidence of the therapeutic effect of ARE. Vehicle group: saline p.o. + saline i.p. ARE group: ARE 20 mg·kg-1·d-1 p.o. + saline i.p. MPTP group: saline p.o. + MPTP 30 mg·kg-1·d-1 i.p. ARE + MPTP group: ARE 20 mg·kg-1·d-1 p.o. + MPTP 30 mg·kg-1·d-1 i.p. ARE: Atractylodis Rhizoma Alba extract; p.o., per oral; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; OFT: open field test. Statistical analyses were measured using one-way analysis of variance. Data are presented as mean ± standard error of mean (n = 7). aP < 0.01 vs the vehicle-treated control group; bP < 0.05 vs the vehicle-treated MPTP-induced PD mice group.

Figure 2 Histological examination evaluating the effect of ARE in a Parkinson’s disease murine model A: immunostaining for TH-positive dopaminergic neurons in the SN of vehicle group (A1), MPTP group (A2), and ARE + MPTP group (A3); B: immunostaining for TH-positive dopaminergic neurons in the striatum of vehicle group (B1), MPTP group (B2), and ARE + MPTP group (B3); C: bar graph representing TH-positive cells in the SN; D: bar graph representing TH-positive intensity in the striatum. Vehicle group: saline p.o. + saline i.p. ARE group: ARE 20 mg·kg-1·d-1 p.o. + saline i.p. MPTP group: saline p.o. + MPTP 30 mg/kg/day i.p. ARE + MPTP group: ARE 20 mg·kg-1·d-1 p.o. + MPTP 30 mg·kg-1·d-1 i.p. ARE: Atractylodis Rhizoma Alba extract; TH: tyrosine hydroxylase; SN: substantia nigra; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Statistical analyses were measured using one-way analysis of variance. Data are presented as mean ± standard error of mean (n = 3). aP < 0.05 vs the control group; bP < 0.05 vs the vehicle-treated MPTP-induced PD mice group.

Figure 3 RNA-seq correlation analysis A: scatter plot; B: heatmap of differentially expressed genes between the MPTP-induced Parkinson’s disease group and ARE-treated MPTP groups (n = 2 mice per group). MPTP group: saline p.o. + MPTP 30 mg·kg-1·d-1 i.p. ARE + MPTP group: ARE 20 mg·kg-1·d-1 p.o. + MPTP 30 mg·kg-1·d-1 i.p. MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; ARE: Atractylodis Rhizoma Alba extract.

Figure 4 Methyl-seq correlation analysis A: the stacking bar graph shows the percentage of hypermethylated and hypomethylated CpGs for each chromosome; B: heamap of differentially methylated DNA between the MPTP-induced Parkinson’s disease group and ARE-treated MPTP groups (n = 2 mice per group). MPTP group: saline p.o. + MPTP 30 mg·kg-1·d-1 i.p. ARE + MPTP group: ARE 20 mg·kg-1·d-1 p.o. + MPTP 30 mg·kg-1·d-1 i.p. MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; ARE: Atractylodis Rhizoma Alba extract.