Journal: Journal of Lipid Research

Article Title: Therapeutic siRNA targeting PLIN2 ameliorates steatosis, inflammation, and fibrosis in steatotic liver disease models

doi: 10.1016/j.jlr.2024.100635

Figure Lengend Snippet: Plin2 is a potential therapeutic target for MASLD. A: RT-PCR analysis of relative mRNA levels of hepatic Plin2 in ob / ob mice and C57/BL6J mice subjected to HFD, MCD, or HFD/HFr diet. B: Schematic representation of the procedure for HFD-induced MASLD in spCas9 KI mice subjected to AAV-mediated gene editing. C: Immunoblotting analysis of hepatic PLIN2 protein levels in HFD-induced MASLD mice subjected to AAV-mediated gene editing. D: RT-PCR analysis of relative mRNA levels of hepatic Plin2 in HFD-induced MASLD mice subjected to AAV-mediated gene editing. E,F: Body weight (E) and liver-to-body weight ratio (F) for HFD-induced MASLD mice subjected to AAV-mediated gene editing. G: Histological analysis of liver sections from HFD-induced MASLD mice subjected to AAV-mediated gene editing. Quantification of Oil red O staining–positive areas and TUNEL-positive cells is shown. Scale bar, 50 μm. H-J: Levels of hepatic TG (H), serum TG (I), and serum TC (J) in HFD-induced MASLD mice subjected to AAV-mediated gene editing. K: Levels of serum ALT in HFD-induced MASLD mice subjected to AAV-mediated gene editing. Data are shown as mean ± SEM. For all panels, n = 6 mice per group. P values were determined using a two-tailed Student's t test. “ns” denotes no significant difference. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. AAV, adeno-associated virus; ALT, alanine aminotransferase; HFD, high-fat diet; HFr, high fructose; KI, knockin; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine-choline deficient; TC, total cholesterol; TG, triglyceride.

Article Snippet: The HFD-induced MASLD mice were administered 1 mg/kg lomitapide (MedChemExpress; Cat# HY-14667) once daily via oral gavage at the beginning of the dark cycle.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Staining, TUNEL Assay, Two Tailed Test, Virus, Knock-In

Journal: Journal of Lipid Research

Article Title: Therapeutic siRNA targeting PLIN2 ameliorates steatosis, inflammation, and fibrosis in steatotic liver disease models

doi: 10.1016/j.jlr.2024.100635

Figure Lengend Snippet: Treatment with GalNAc-si Plin2 ameliorates MASLD induced by HFD. A: Schematic representation of the HFD-induced MASLD in C57BL/6J mice subcutaneously injected with 4 mg/kg GalNAc-si Plin2 , GalNAc-siNC, or PBS. Mice fed with a chow diet serve as the healthy control. B: The liver-to-body weight ratio for chow diet-fed mice and HFD-induced MASLD mice subcutaneously injected with 4 mg/kg GalNAc-si Plin2 , GalNAc-siNC, or PBS. C: Histological analysis of liver sections from chow diet-fed mice and HFD-induced MASLD mice subcutaneously injected with 4 mg/kg GalNAc-si Plin2 , GalNAc-siNC, or PBS. Quantification of Oil red O staining–positive areas and TUNEL-positive cells is shown. Scale bar, 50 μm. D-G: Levels of hepatic TG (D), serum TG (E), serum TC (F), and serum ALT (G) from chow diet–fed mice and HFD-induced MASLD mice subcutaneously injected with 4 mg/kg GalNAc-si Plin2 , GalNAc-siNC, or PBS. H: Schematic illustration of the GTT and ITT experimental procedures. I,J: Assessment of glucose tolerance and insulin sensitivity in HFD-induced MASLD mice subcutaneously injected with 4 mg/kg GalNAc-si Plin2 , GalNAc-siNC, or PBS. The incremental area under the curve (iAUC0-120 min) was quantified. Data are shown as mean ± SEM. n = 6 mice per group for panels (A–G). n = 5 mice per group for panels (I and J). P values were determined using one-way ANOVA followed by Tukey's multiple comparison tests. “ns” denotes no significant difference. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. ALT, alanine aminotransferase; GalNAc, N-acetylgalactosamine; GTT, glucose tolerance test; HFD, high-fat diet; ITT, insulin tolerance test; MASLD, metabolic dysfunction–associated steatotic liver disease; TC, total cholesterol; TG, triglyceride.

Article Snippet: The HFD-induced MASLD mice were administered 1 mg/kg lomitapide (MedChemExpress; Cat# HY-14667) once daily via oral gavage at the beginning of the dark cycle.

Techniques: Injection, Control, Staining, TUNEL Assay, Comparison

Journal: Journal of Lipid Research

Article Title: Therapeutic siRNA targeting PLIN2 ameliorates steatosis, inflammation, and fibrosis in steatotic liver disease models

doi: 10.1016/j.jlr.2024.100635

Figure Lengend Snippet: Treatment with GalNAc-si Plin2 increases the hepatic secretion of VLDL TG. A: Treatment with GalNAc-si Plin2 increased the hepatic secretion of VLDL TG. B: Immunoblotting of hepatic MTP in HFD-induced MASLD mice subcutaneously injected with 4 mg/kg GalNAc-si Plin2 , GalNAc-siNC, or PBS. C,D: The body weights (C) and the ratios of liver weight to body weight (D) in HFD-induced MASLD mice treated with lomitapide, followed by subcutaneous injection of 4 mg/kg GalNAc-si Plin2 , GalNAc-siNC, or PBS. E: Blocking hepatic secretion of VLDL TG by lomitapide abolished the effects of GalNAc-si Plin2 on hepatic steatosis and liver injury. Quantification of Oil red O staining–positive areas and TUNEL-positive cells are shown. Scale bar, 50 μm. F-I: Levels of hepatic TG (F), serum TG (G), serum TC (H), and serum ALT (I) in HFD-induced MASLD mice treated with lomitapide, followed by subcutaneous injection of 4 mg/kg GalNAc-si Plin2 , GalNAc-siNC, or PBS. J-L: Oxygen consumption (VO 2 ) (J), carbon dioxide production (VCO 2 ) (K), and energy expenditure (EE) (L) in mice treated with 4 mg/kg GalNAc-si Plin2 , GalNAc-siNC, or PBS at 4°C. Data are shown as mean ± SEM. For panel (A), n = 4 mice per group, and P values were determined using two-way ANOVA followed by Bonferroni's multiple comparisons test. For panels (C–I), n = 5 mice per group, and P values were calculated using one-way ANOVA followed by Tukey's multiple comparison tests. For panels (J–L), n = 5 mice per group, and P values were calculated using two-way ANOVA followed by Bonferroni's multiple comparisons test or one-way ANOVA followed by Tukey's multiple comparison tests. “ns” denotes no significant difference. ∗∗∗ P < 0.001. ALT, alanine aminotransferase; AST, aspartate aminotransferase; GalNAc, N-acetylgalactosamine; HFD, high-fat diet; MASLD, metabolic dysfunction–associated steatotic liver disease; TC, total cholesterol; TG, triglyceride.

Article Snippet: The HFD-induced MASLD mice were administered 1 mg/kg lomitapide (MedChemExpress; Cat# HY-14667) once daily via oral gavage at the beginning of the dark cycle.

Techniques: Western Blot, Injection, Blocking Assay, Staining, TUNEL Assay, Comparison