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. 2021 Jun 7;17(1):170–177. doi: 10.4103/1673-5374.314314

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Copyright: © Neural Regeneration Research

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Ghrelin modulates α-synuclein expression by inhibiting lincRNA-p21 expression and preserving TGIF1 levels.

(A) Representative western blot assay and quantification of TGIF1 in SH-SY5Y cells pretreated with different concentrations of ghrelin for 30 minutes followed by incubation with 6-OHDA for 24 hours. (B) Results of the dual-luciferase reporter assay. (C) qRT-PCR analysis of TGIF1 mRNA levels in SH-SY5Y cells transfected with TGIF1 siRNA for 24 hours. (D) qRT-PCR analysis of SNCA mRNA levels in SH-SY5Y cells transfected with TGIF1 siRNA and treated with 6-OHDA with or without the presence of 1 nM ghrelin for 24 hours. (E) qRT-PCR analysis of lincRNA-p21 levels in SH-SY5Y cells transfected with pcDNA-p21 for 24 hours. (F) qRT-PCR analysis of TGIF1 mRNA levels in SH-SY5Y cells transfected with pcDNA-p21 and treated with 6-OHDA with or without the presence of 1 nM ghrelin for 24 hours. (G) qRT-PCR analysis of SNCA mRNA levels in SH-SY5Y cells transfected with pcDNA-p21 and treated with 6-OHDA with or without the presence of 1 nM ghrelin for 24 hours. (H) Representative western blot and quantification data of α-synuclein and TGIF1 in SH-SY5Y cells transfected with TGIF1 siRNA or pcDNA-p21 and treated with 6-OHDA with or without the presence of 1 nM ghrelin for 24 hours. The data are shown as the mean ± SD (n = 5; one-way analysis of variance followed by Bonferroni post hoc test); *P < 0.05, vs. control group; #P < 0.05, ##P < 0.01, vs. 6-OHDA treatment group; &P < 0.05, &&P < 0.01, vs. NC group; ††P < 0.01, vs. 6-OHDA + 1 nM ghrelin group; ^^P < 0.01, vs. pGL4.10-SNCA promoter (WT) + pcDNA3.1-3FLAG group; $$P < 0.01, vs. pGL4.10-SNCA promoter (MUT) + pcDNA3.1-3FLAG group. 6-OHDA: 6-Hydroxydopamine; Con: control; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; Luc: luciferase; NC: negative control; qRT-PCR: reverse transcription quantitative polymerase chain reaction; R-Luc: Renilla luciferase; siRNA: small interfering RNA; SNCA: the gene encoding α-synuclein; TGIF1: TG-interacting factor 1; α-syn: α-synuclein.