Corrigendum: Dl-3- n-Butylphthalide Exerts Dopaminergic Neuroprotection Through Inhibition of Neuroinflammation

Abstract

[This corrects the article DOI: 10.3389/fnagi.2019.00044.].

Keywords: MAPK; NF-κB; Parkinson's disease; dl-3-n-butylphthalide; microglia; neuroinflammation.

Figure 5

NBP protected dopaminergic neurons from neurotoxicity induced by microglial activation. SH-SY5Y cells were incubated for 24 h with conditioned medium derived from cultures of BV-2 cells. Before collecting culture media, BV-2 cells were pretreated with NBP (0 or 100 μM) for 1 h and incubated with LPS (0 or 100 ng/ml) or MPP⁺ (0 or 500 μM) for 24 h. (A) Cell viability was measured with the CCK8 assay (n = 5). All data are presented as means ± SEM. **p < 0.01, ***p < 0.001, compared with the Control group; ^#p < 0.05, ^##p < 0.01, ^###p < 0.001, compared with the LPS group or the MPP⁺ group. (B) The apoptosis of SH-SY5Y was evaluated by immunofluorescence detection of cleaved caspase-3 (green) and cell nuclei was stained with DAPI (blue) (scale bar: 50 μm). (C) The cell death of SH-SY5Y was evaluated by immunofluorescence detection of PI (red) (scale bar: 50 μm).

Figure 6

NBP reduced pro-inflammatory molecules expression in LPS-stimulated BV-2 cells. BV-2 cells were pretreated with NBP (0 or 100 μM) for 1 h followed by LPS (0 or 100 ng/ml) for 6 h and total RNA was isolated. To measure cellular protein expression or cytokines level in supernatants, time for LPS treatment was 24 h. (A) The mRNA expression of IL-1β, IL-6, TNF-a, iNOS and COX-2 was analyzed by RT-PCR and normalized to that of β-actin. (B) The protein level of TNF-α, IL-1β, iNOS and COX2 was analyzed by Western Blot. (C) The level of TNF-α and IL-1β in supernatants was assayed by ELISA kit. All data are presented as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, compared with the Control group; ^#p < 0.05, ^##p < 0.01, ^###p < 0.001, compared with the LPS group.

Figure 8

NBP inhibited the activation of ERK, NF-κB and PI3K/Akt pathways in LPS-stimulated BV-2 cells. (A) Western Blot assay for MAPK expression. The expression of total JNK/p38/ERK as well as p-JNK/p-p38/p-ERK were analyzed. (B) Western Blot assay for NF-κB expression in whole-cell, nuclear and cytoplasmic extracts. The nuclear translocation of p65 was also evaluated by immunofluorescence detection of p65 (green) and cell nuclei was stained with DAPI (blue) (scale bar: 50 μm). (C) Western Blot assay for PI3K/Akt expression. The expression of total PI3K/Akt as well as p-PI3K/p-Akt were analyzed. All data are presented as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, compared with the Control group; ^#p < 0.05, ^##p < 0.01, ^###p < 0.001, compared with the LPS group.