Table 1.
The primers for real-time PCR.
Fig 1.
Effects of isobavachalcone on motor, balance and coordination abilities of PD mouse.
Isobavachalcone (Iso, 50 mg/kg) significantly prolonged the residence time of mice on Rota-rod. Bars indicate the mean±SD of three independent experiments. ## p<0.01 indicating very significantly different from Control group. * p<0.05 indicating statistically significantly different from MPTP injury group.
Fig 2.
Effects of isobavachalcone on microglia, astrocytes and neurons.
(A) Brain sections were immunostained for TH immunoreactivity in SNpc. The activation of microglia and astrocytes respectively were detected by Iba-1 and GFAP immunostaining (figs C and E). The number of TH, Iba-1 and GFAP positive cells were counted, and the data were expressed as mean±SD (figs B, D and F). Those results show that isobavachalcone (Iso, 50 mg/kg) could inhibit either the elevated levels of Iba-1 and GFAP, or the necrosis of neurons. Images were obtained from the SNpc of Iba-1, TH and GFAP. Scale bars: 200μm and 500μm in A, and 50μm in C and E, n = 4. ## p<0.01 indicating very significantly different from Control group. ** p<0.01 indicating very significantly different from MPTP injury group.
Fig 3.
Changes of microglia inflammatory cytokines in vitro and in vivo.
A: Isobavachalcone (Iso) significantly inhibited the transcription of IL-6 and IL-1β induced by MPTP injury. B-E: Isobavachalcone inhibited the transcriptional levels of TNF-α, IL-6, IL-1β, and IL-10 induced by LPS in BV-2 cells. F: The expression levels of both TNF-α and IL-6 were decreased with the increasing of the administration isobavachalcone concentration. Bars indicate the mean±SD. ##,$ $ p<0.01 vs. respective control group. *,& p<0.05, **p<0.01, ***,&&& p<0.001 vs. PD model or LPS-treated group.
Fig 4.
Effects of isobavachalcone treatment on NO and iNOS.
(A, B) Isobavachalcone (Iso) significantly inhibited the levels of NO and the transcriptional levels of iNOS in LPS-treated BV-2 cells. C: Effect of isobavachalcone treatment on iNOS mRNA transcriptional levels in PD mouse brain. Bars indicate the mean±SD of three independent experiments. ##p<0.01 vs. Control group. ** p<0.01 and *** p<0.001 vs. LPS-treated group.
Fig 5.
Effect of isobavachalcone on NF-κB pathway.
(A1, A2, B1, and B2) The expression levels of p65 in BV-2 cells and the mouse brain tissue. (C and D) Isobavachalcone (Iso) inhibited the DNA binding activity (C) and nuclear transfer (D) of NF-κB p65. Bars indicate the mean±SD of three independent experiments. # p<0.05, ## p<0.01 in comparison to Control group. * p<0.05, ** p<0.01 in comparison to LPS group.
Fig 6.
Effects of isobavachalcone treatment on BV-2 cells and Neuro-2a cells.
A: Isobavachalcone (Iso) caused no significant cytotoxicity to BV-2 cells. B: The effect of isobavachalcone treated BV-2 cells supernatants on Neuro-2a cells. BV-2 cells CM was treated by LPS-treated together with isobavachalcone (LPS+Iso group), exerting a protective effect on Neuro-2a cells. And such protective effect was stronger than that of the directly treatment with isobavachalcone plusing the supernatant from LPS-treated BV-2 cells (LPS/Iso group). Means: Control (DMEM basic culture medium), LPS (supernatant from LPS treated BV-2 cells), LPS+Iso (supernatant from LPS plus isobavachalcone treated BV-2 cells), LPS/Iso (isobavachalcone (final concentration 5μM) dissolved in supernatant from LPS treated BV-2 cells). C: Isobavachalcone decreased microglial-induced neuro-2a death in a co-culture system. Bars indicated the mean±SD of three independent experiments. ## p<0.01 in comparison to control group. * p<0.05, ** p<0.01 in comparison to LPS group. △△△ p<0.001 in comparison to LPS+Iso group (supernatant from LPS plus isobavachalcone treated BV-2 cells).