Fig. 1
The liganans, including JA, JB, and JC, isolated from PJ exert neuroprotective effect in nerve cell, C. elegans, and rat models of PD. a Pictures of the original plant and dried herbal piece of Peristrophe japonica, and the chemical structures of JA, JB, and JC. b Representative images of cells with GFP or GFP-LC3 puncta, and the average number of GFP-LC3 puncta per cell in JA, JB, or JC treated stable RFP-GFP-LC3 U87 cells at 24 h (scale bar: 50 μm). c Protein expression and quantification of LC3-II in JA, JB, or JC treated PC-12 and SHSY5Y cells at 24 h. d Protein expression of the phosphorylation and total forms of AMPK, mTOR, ULK1, P70s6K, and 4EBP1 in JA, JB, JC, or Rapamycin (Rap) treated PC-12 and SHSY5Y cells at 24 h. e Representative images of cells with GFP or GFP-LC3 puncta, and the average number of GFP-LC3 puncta per cell in stable RFP-GFP-LC3 U87 cells treated with JA (0.13 μM), JB (0.13 μM), or JC (4 μM) in the presence or absence of CC (2.5 μM), SCH (10 μM), LY (25 μM), or Baf (1 nM) for 24 h (scale bar: 50 μm). f Upper: representative images of cells with GFP or GFP-LC3 puncta, and the average number of GFP-LC3 puncta per cell in GFP-LC3 transiently transfected MEF Atg7−/− and MEF Atg7+/+ cells. Below: protein expression of LC3-II in JA, JB, or JC treated MEF Atg7−/− and MEF Atg7+/+ cells at 24 h (scale bar: 50 μm). g Cell viability of 6-OHDA-induced PC-12 and SHSY5Y cells treated with JA, JB, or JC for 24 h was measured by MTT assay. h ROS production of 6-OHDA-induced PC-12 cells or SHSY5Y cells treated with JA (0.13 μM), JB (0.13 μM), JC (4 μM), or NAC (1 mM) for 24 h was measured by flow cytometer using H2DCFDA reagent. i Representative images and quantification of p62 and GFP-LGG-1 puncta in BC12921 and DA2123 C. elegans strains treated with JA, JB, or JC (scale bar: 200 or 100 μm). j GFP expression in dopaminergic neurons of 6-OHDA-induced BZ555 strain and YFP-α-synuclein expression of NL5901 strain treated with L-Dopamine (L-Dopa), JA, JB, or JC (scale bar: 100 μm). k Motor performances including net number of rotation in 30 min, average swimming score, and average forelimb hanging score of 6-OHDA rats administrated with L-Dopa or JA. l Protein expression and quantification of TH and α-synuclein in brain corpus striatum of 6-OHDA-induced rats treated with L-Dopa or JA. All the experiments were performed in triplicates and data represented the mean ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001)
