Illustrations
Figure 1
lncRNA NEAT1 knockdown remitted NAFLD in the FFA-induced NAFLD cellular model. Concentrations of TG were detected by ELISA in the FFA-induced NAFLD cellular model. B. Relative expression of NEAT1 in FFA-induced BRL3A cells transfected with NEAT1 siRNA and siRNA-NC was assessed by qRT-PCR, normalized to GAPDH. C. CCK-8 assay was performed to assess cell proliferation in FFA-induced BRL3A cell transfected with NEAT1 siRNA and siRNA-NC. D. Relative expression of molecular markers of NAFLD (FAS and ACC) in FFA-induced BRL3A cell transfected with NEAT1 siRNA and siRNA-NC was assessed by qRT-PCR, normalized to GAPDH. E. Protein levels of FAS and ACC in FFA-induced BRL3A cell transfected with NEAT1 siRNA and siRNA-NC were assessed by Western blot. F. Relative expression of two fibrosis factors α-SMA and Collagen I in FFA-induced BRL3A cells transfected with NEAT1 siRNA and siRNA-NC was assessed by qRT-PCR, normalized to GAPDH. G. Relative expression of inflammatory cytokines (TNF-α, IL-1β, IL-6, and MCP1) in FFA-induced BRL3A cells transfected with NEAT1 siRNA and siRNA-NC was assessed by qRT-PCR, normalized to GAPDH. H. Immunofluorescence staining of two fibrosis factors α-SMA and Collagen I in FFA-induced BRL3A cells transfected with NEAT1 siRNA and siRNA-NC. *P < 0.05 and **P < 0.01. All data were from three independent experiments performed in triplicate.
Figure 1
Figure 2
lncRNA NEAT1 knockdown inhibited GLI3 expression and promoted miR-506 expression. A. Protein levels of core transcription factors SMO, GLI2, and GLI3 in Hh signaling pathway in FFA-induced BRL3A cell transfected with NEAT1 shRNA and shRNA-NC were assessed by Western blot, normalized to GAPDH. B. Relative expression of miR-506 in FFA-induced BRL3A cell transfected with NEAT1 shRNA and shRNA-NC was assessed by qRT-PCR, normalized to U6. C. The dual-luciferase activity of the NEAT1-WT and NEAT1-MUT in BRL3A cells transfected with miR-506 mimics. D. Predicted binding site between miR-506 and NEAT1 by starbase software. *P < 0.05 and **P < 0.01. All data were from three independent experiments performed in triplicate.
Figure 2
Figure 3
miR-506 overexpression remitted NAFLD in the FFA-induced NAFLD cellular model. A. Relative expression of miR-506 in FFA-induced BRL3A cells transfected with miR-506 mimics and mimics-NC was assessed by qRT-PCR, normalized to U6. B. CCK-8 assay was performed to assess cell proliferation in FFA-induced BRL3A cell transfected with miR-506 mimics and mimics-NC. C. Relative expression of molecular markers of NAFLD (FAS and ACC) in FFA-induced BRL3A cell transfected with miR-506 mimics and mimics-NC was assessed by qRT-PCR, normalized to GAPDH. D. Protein levels of FAS and ACC in FFA-induced BRL3A cell transfected with miR-506 mimics and mimics-NC were assessed by Western blot. E. Relative expression of two fibrosis factors α-SMA and Collagen I in FFA-induced BRL3A cells transfected with miR-506 mimics and mimics-NC was assessed by qRT-PCR, normalized to GAPDH. F. Immunofluorescence staining of two fibrosis factors α-SMA and Collagen I in FFA-induced BRL3A cells transfected with miR-506 mimics and NC mimics. *P < 0.05 and **P < 0.01. All data were from three independent experiments performed in triplicate. G. Relative expression of inflammatory cytokines (TNF-α, IL-1β, IL-6, and MCP1) in FFA-induced BRL3A cells transfected with miR-506 mimics and mimics-NC was assessed by qRT-PCR, normalized to GAPDH.
Figure 3
Figure 4
miR-506 overexpression inhibited Hh signaling pathway in the FFA-induced NAFLD cell model. A. Protein levels of core transcription factors SMO, GLI2, and GLI3 in Hh signaling pathway in FFA-induced BRL3A cell transfected with miR-506 mimics and mimics-NC were assessed by Western blot normalized to GAPDH. B. Relative expression of lncRNA NEAT1 in FFA-induced BRL3A cell transfected with miR-506 mimics and mimics-NC was assessed by qRT-PCR, normalized to U6. C. The dual-luciferase activity of the GLI3-WT and GLI3-MUT in BRL3A cells transfected with miR-506 mimics. D. Predicted binding site between miR-506 and GLI3 by targetscan software. *P < 0.05 and **P < 0.01. All data were from three independent experiments performed in triplicate.
Figure 4
Figure 5
lncRNA NEAT1 regulated NAFLD through sponging miR-506. Relative expression of two fibrosis factors α-SMA and Collagen I in FFA-induced BRL3A cell transfected with miR-506 inhibitor or inhibitor-NC after lncRNA NEAT1 knockdown were assessed by qRT-PCR, normalized to GAPDH. B. Relative expression of inflammatory cytokines (TNF-α, IL-1β, IL-6 and MCP1) in FFA-induced BRL3A cells transfected with miR-506 inhibitor or inhibitor-NC after lncRNA NEAT1 knockdown were assessed by qRT-PCR, normalized to GAPDH. C. Protein level of FAS and ACC in FFA-induced BRL3A cell transfected with miR-506 inhibitor or inhibitor-NC after lncRNA NEAT1 knockdown were assessed by Western blot. D. Immunofluorescence staining of two fibrosis factors α-SMA and Collagen I in FFA-induced BRL3A cells transfected with miR-506 inhibitor or inhibitor-NC after NEAT1 knockdown. *P < 0.05 and **P < 0.01. All data were from three independent experiments performed in triplicate.
Figure 5
Auteurs
1 Department of Gastroenterology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang Province, P.R China
2 Department of Pharmacy, Wenzhou People's Hospital, Wenzhou 325000, Zhejiang Province, P.R China
* Corresponding author: Dr. Hua-Qin Guan, Department of Gastroenterology, The First Affiliated Hospital of Wenzhou Medical University, New Campus of Nanbaixiang Wenyi First Hospital, Ouhai District, Wenzhou 325000, Zhejiang Province, P.R China
Background As one of the most common liver disorders worldwide, nonalcoholic fatty liver disease (NAFLD) begins with the abnormal accumulation of triglyceride (TG) in the liver and can lead to inflammation and fibrosis. Long noncoding RNA (lncRNA) NEAT1 was reported to promote NAFLD progress. However, its molecular mechanism in NAFLD was not fully clear.