t2dm sdf1 neutralising antibody  (R&D Systems)

Journal: Journal of Cellular and Molecular Medicine

Article Title: Stromal Cell Derived Factor‐1 Promotes Hepatic Insulin Resistance via Inhibiting Hepatocyte Lipophagy

doi: 10.1111/jcmm.70352

Figure Lengend Snippet: The neutralising of SDF‐1 mitigates hepatic IR via promoting hepatocyte lipophagy in T2DM mice. The mice were assigned into normal, T2DM, T2DM + SDF‐1 neutralising antibody (1 mg/kg, intrahepatic injection), T2DM + SDF‐1 neutralising antibody +3‐MA (30 mg/kg, intrahepatic injection), and T2DM + MET (250 mg/kg/day, oral administration) groups. (A) The experimental flow was shown. (B) Hepatocyte marker albumin (green) and SDF‐1 (red) were labelled in mouse liver tissues. (C) The co‐localization of albumin and SDF‐1 was analysed. (D) Fasting blood insulin levels were detected. (E) Fasting blood glucose levels were detected. (F) HOMA‐IR was calculated. (G) The liver index was analysed. (H) Representative images of mouse livers were shown. (I) Liver oil red O staining was shown. (J) The autophagy marker LC3 (green) was labelled in mouse liver tissues. (K) The puncta LC3 per cell was analysed. ** p < 0.01, *** p < 0.001 versus normal group. # p < 0.05，## p < 0.01，### p < 0.001 versus T2DM group. ** p < 0.01 versus T2DM + SDF‐1 neutralising antibody group. The n.s. stood for no significance.

Article Snippet: The mice were assigned into normal, T2DM, T2DM + SDF1 neutralising antibody (MAB310, R&D Systems, USA; 1 mg/kg/day, intrahepatic injection), T2DM + SDF1 neutralising antibody +3‐Methyladenine (3‐MA; autophagy inhibitor; HY‐19312, MedChemExpress, USA; 30 mg/kg/day, intrahepatic injection), T2DM + metformin (MET; S5958, Selleck, USA; 250 mg/kg/day, once a day from week 4 to week 6 of HFHSD feeding, oral administration) groups.

Techniques: Injection, Marker, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Stromal Cell Derived Factor‐1 Promotes Hepatic Insulin Resistance via Inhibiting Hepatocyte Lipophagy

doi: 10.1111/jcmm.70352

Figure Lengend Snippet: SDF‐1 expression and release increase in PA‐treated hepatocytes. The hepatocytes were divided into normal and PA (0.5 mmol/L for 24 h) groups. (A) SDF1 protein levels in hepatocytes were detected by Western blot. (B) SDF‐1 relative protein levels were analysed. (C) SDF‐1 protein levels in hepatocyte culture supernatant were measured by ELISA. ** p < 0.01, *** p < 0.001 versus normal group.

Article Snippet: The mice were assigned into normal, T2DM, T2DM + SDF1 neutralising antibody (MAB310, R&D Systems, USA; 1 mg/kg/day, intrahepatic injection), T2DM + SDF1 neutralising antibody +3‐Methyladenine (3‐MA; autophagy inhibitor; HY‐19312, MedChemExpress, USA; 30 mg/kg/day, intrahepatic injection), T2DM + metformin (MET; S5958, Selleck, USA; 250 mg/kg/day, once a day from week 4 to week 6 of HFHSD feeding, oral administration) groups.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Stromal Cell Derived Factor‐1 Promotes Hepatic Insulin Resistance via Inhibiting Hepatocyte Lipophagy

doi: 10.1111/jcmm.70352

Figure Lengend Snippet: SDF‐1 inhibits lipophagy in PA‐treated hepatocytes via CXCR4, rather than CXCR7. The hepatocytes were divided into normal, PA, PA + SDF‐1 neutralising antibody (1 μg for 24 h), PA + AMD3100 (10 μM for 24 h), and PA + ACT‐1004‐1239 (6 nM for 24 h) groups. (A) LC3, p62 and ATG7 protein levels in hepatocytes were detected by Western blot. (B–D) LC3, p62 and ATG7 relative protein levels were analysed. (E) The co‐localization of the autolysosome (red) and the LD (green) in hepatocytes, indicated the induction of lipophagy (yellow). (F) The co‐localization of autolysosome and LD was analysed. (G) The LD inside the hepatocytes was visualised by oil red O staining. ** p < 0.01, *** p < 0.001 versus normal group. # p < 0.05, ## p < 0.01 versus PA group.

Article Snippet: The mice were assigned into normal, T2DM, T2DM + SDF1 neutralising antibody (MAB310, R&D Systems, USA; 1 mg/kg/day, intrahepatic injection), T2DM + SDF1 neutralising antibody +3‐Methyladenine (3‐MA; autophagy inhibitor; HY‐19312, MedChemExpress, USA; 30 mg/kg/day, intrahepatic injection), T2DM + metformin (MET; S5958, Selleck, USA; 250 mg/kg/day, once a day from week 4 to week 6 of HFHSD feeding, oral administration) groups.

Techniques: Western Blot, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Stromal Cell Derived Factor‐1 Promotes Hepatic Insulin Resistance via Inhibiting Hepatocyte Lipophagy

doi: 10.1111/jcmm.70352

Figure Lengend Snippet: SDF‐1/CXCR4 inhibits lipophagy in PA‐treated hepatocytes via activating AKT/mTOR pathway. The hepatocytes were divided into normal, PA, PA + SDF‐1 neutralising antibody, PA + AMD3100, PA + MK‐2206 2HCl (10 μM for 24 h), and PA + XL388 (100 nM for 24 h) groups. (A) P‐AKT, AKT, p‐mTOR and mTOR protein levels in hepatocytes were detected by Western blot. (B, C) P‐AKT and p‐mTOR relative protein levels were analysed. In Figure , ** p < 0.01, *** p < 0.001 versus normal group. # p < 0.05, ## p < 0.01 versus PA group. The hepatocytes were divided into PA + SDF‐1 neutralising antibody, PA + SDF‐1 neutralising antibody + SC79 (4 μg/mL for 24 h), PA + SDF‐1 neutralising antibody + MHY1485 (5 μM for 24 h), PA + AMD3100, PA + AMD3100 + SC79 and PA + AMD3100 + MHY1485 groups. (D) LC3, p62 and ATG7 protein levels in hepatocytes were detected by Western blot. (E–G) LC3, p62 and ATG7 relative protein levels were analysed. (H) The co‐localization of the autolysosome (red) and the LD (green) in hepatocytes, indicated the induction of lipophagy (yellow). (I) The co‐localization of autolysosome and LD was analysed. (J) The LD inside the hepatocytes was visualised by oil red O staining. In Figure , ** p < 0.01, *** p < 0.001 versus PA + SDF‐1 neutralising antibody group. ## p < 0.01, ### p < 0.001 versus PA + AMD3100 group.

Article Snippet: The mice were assigned into normal, T2DM, T2DM + SDF1 neutralising antibody (MAB310, R&D Systems, USA; 1 mg/kg/day, intrahepatic injection), T2DM + SDF1 neutralising antibody +3‐Methyladenine (3‐MA; autophagy inhibitor; HY‐19312, MedChemExpress, USA; 30 mg/kg/day, intrahepatic injection), T2DM + metformin (MET; S5958, Selleck, USA; 250 mg/kg/day, once a day from week 4 to week 6 of HFHSD feeding, oral administration) groups.

Techniques: Western Blot, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Stromal Cell Derived Factor‐1 Promotes Hepatic Insulin Resistance via Inhibiting Hepatocyte Lipophagy

doi: 10.1111/jcmm.70352

Figure Lengend Snippet: SDF1/CXCR4/AKT/mTOR pathway‐inhibited lipophagy promotes PA‐induced hepatocyte IR. The hepatocytes were divided into normal, PA, PA + SDF‐1 neutralising antibody, PA + SDF‐1 neutralising antibody +3‐MA (10 mM for 24 h), PA + AMD3100, PA + AMD3100 + 3‐MA, PA + MK‐2206 2HCl, PA + MK‐2206 2HCl + 3‐MA, PA + XL388 and PA + XL388 + 3‐MA groups. (A) The glucose content of the medium was measured. (B) Glucose consumption by hepatocytes was analysed. ** p < 0.01 versus normal group. ## p < 0.01 versus PA group. @ p < 0.05 versus PA + SDF‐1 neutralising antibody group. $ p < 0.05 versus PA + AMD3100 group. % p < 0.05 versus PA + MK‐2206 2HCl group. & p < 0.05 versus PA + XL388 group.

Article Snippet: The mice were assigned into normal, T2DM, T2DM + SDF1 neutralising antibody (MAB310, R&D Systems, USA; 1 mg/kg/day, intrahepatic injection), T2DM + SDF1 neutralising antibody +3‐Methyladenine (3‐MA; autophagy inhibitor; HY‐19312, MedChemExpress, USA; 30 mg/kg/day, intrahepatic injection), T2DM + metformin (MET; S5958, Selleck, USA; 250 mg/kg/day, once a day from week 4 to week 6 of HFHSD feeding, oral administration) groups.

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Stromal Cell Derived Factor‐1 Promotes Hepatic Insulin Resistance via Inhibiting Hepatocyte Lipophagy

doi: 10.1111/jcmm.70352

Figure Lengend Snippet: The role and mechanism of SDF‐1 in hepatic IR were displayed. Up‐regulated SDF1 binds to its receptor CXCR4 and CXCR7 on hepatocytes following PA treatment. SDF‐1/CXCR4 signalling, not SDF‐1/CXCR7 signalling, inhibits lipophagy in hepatocytes via activating the phosphorylation of AKT and mTOR1 to promote PA‐induced IR. The blockade of SDF‐1/CXCR4/AKT/mTOR signalling‐induced lipophagy alleviates IR in PA‐treated hepatocytes.

Article Snippet: The mice were assigned into normal, T2DM, T2DM + SDF1 neutralising antibody (MAB310, R&D Systems, USA; 1 mg/kg/day, intrahepatic injection), T2DM + SDF1 neutralising antibody +3‐Methyladenine (3‐MA; autophagy inhibitor; HY‐19312, MedChemExpress, USA; 30 mg/kg/day, intrahepatic injection), T2DM + metformin (MET; S5958, Selleck, USA; 250 mg/kg/day, once a day from week 4 to week 6 of HFHSD feeding, oral administration) groups.

Techniques: Phospho-proteomics