Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: HAL hepatotoxicity and the induced inflammatory response depend on ovarian hormones. (A) For each mouse, the stage of estrous cycle was determined by vaginal cytology analysis before treatment with HAL (5 mmol/kg, ip). Plasma ALT activity was measured 12 h after HAL administration (n = 3–5 per group). P, proestrus; E/M, estrus/metestrus; D, diestrus. *significantly different from other groups. (B) Plasma ALT activity was evaluated 12 h after vehicle (VEH) or HAL (15 mmol/kg, ip) administration in OVX or SHAM mice (n = 3–5 per group). *significantly different from respective VEH control; #significantly different from HAL-treated SHAM mice. (C) Plasma IFN-γ concentration was evaluated in at various times after HAL administration (n = 5–6 per group). #significantly different from time-matched male group; *significantly different from sex-matched 6 h group. (D) IFN-γ concentration was evaluated 12 h after HAL treatment in SHAM and OVX mice (n = 4 per group). *significantly different from SHAM group.

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques: Activity Assay, Concentration Assay

Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: IFN-γ KO mice are protected from developing severe HAL hepatotoxicity. Female WT BALB/cJ (WT) and IFN-γ KO mice were treated with HAL (15 mmol/kg, ip), and plasma and liver samples were collected at various times. (A) Plasma ALT activity was evaluated 8 and 12 h after HAL treatment (n = 5–6 per group). *significantly different from HAL-treated WT mice. (B) Immunoblot detection of TFA protein adducts in liver homogenates (n = 3 per group). (C) Hematoxylin and eosin liver sections from HAL-treated WT and IFN-γ KO mice 30 h after treatment. Labeled in picture are central vein (CV) and portal triad (PT). Images were photographed at ×200 magnification.

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques: Activity Assay, Western Blot, Labeling

Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: HMGB-1 release and the response to HAL in Tlr4Lps-d mice. (A) Plasma HMGB-1 concentration was evaluated at various times after HAL treatment (15 mmol/kg, ip) in male and female mice (n = 6 per group). VEH-treated animals had plasma HMGB1 concentrations < 5 pg/ml. #significantly different from sex-matched 6 h time point. *significantly different from time-matched male and all other female groups. (B and C) Female WT BALB/cBYJ (WT) mice and Tlr4Lps-d mice were treated with HAL (15 mmol/kg, ip). Plasma ALT activity and IFN-γ concentration were evaluated 24 h after HAL treatment (n = 4–5 per group). *significantly different from WT controls.

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques: Concentration Assay, Activity Assay

Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: KC-depleted mice and CD1d KO and RAGNULL mice develop severe HAL-induced liver injury. Control- or clodronate liposome-pretreated mice were given VEH or HAL (15 mmol/kg, ip), and plasma and liver samples were collected 24 h later. Plasma ALT activity (A) and IFN-γ concentration (B) were evaluated (n = 4–6 per group). WT BALB/cJ (WT), NKT-deficient mice (CD1d KO), or T- and B cell–deficient mice (RAGNULL) were treated with HAL (15 mmol/kg, ip). (C) Plasma ALT activity was evaluated in WT and CD1d KO mice 12 and 24 h after HAL administration (n = 5 per group). (D) Plasma ALT activity was evaluated in HAL-treated WT and RAGNULL mice 12 h after HAL administration (n = 5 per group). (E and F) Hematoxylin and eosin liver sections taken 24 h after HAL treatment of CD1d KO and RAGNULL mice. Labeled in picture are central vein (CV) and portal triad (PT). Arrowheads identify areas of necrosis. Images were photographed at ×200X magnification.

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques: Activity Assay, Concentration Assay, Labeling

Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: HAL-induced hepatotoxicity depends on NK cell activity. Mice treated with IgG or anti-AsGM1 were given HAL (15 mmol/kg, ip) as described in Methods section, and plasma samples were collected at 12 and 24 h. Plasma ALT activity (A) and IFN-γ concentration (B) (n = 4–6 per group). *significantly different from time-matched controls. Plasma ALT activity (C) and IFN-γ concentration (D) in WT and BALBPrf1 mice 12 h after HAL administration (n = 4 per group). *significantly different from WT mice.

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques: Activity Assay, Concentration Assay

Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: Proposed mechanism of innate immune-mediated severe HAL-induced liver injury. (A) In the absence of stress stimulation, self-proteins such as H2Dd are expressed on the plasma membrane of hepatocytes and RAE-1 is not; this condition keeps NK cells quiescent. (B) When hepatocytes are exposed to HAL, intracellular TFA adducts form (1). This induces a stress response in hepatocytes (2) that alters the surface NK receptor ligands (3) and activates NK cells. Activated NK cells release IFN-γ as well as the contents of cytotoxic granules (4), such as perforin and granzyme B, which contribute to hepatocellular necrosis (5). Damaged hepatocytes release endogenous danger signals, such as HMGB-1 (6). These endogenous danger signals are ligands for TLR4 (7), and the resultant signals are involved in a positive feedback loop that further activates NK cells as well as recruits polymorphonuclear leukocytes (PMNs) (Scaffidi et al., 2002) that participate (8) in the progression of injury (Dugan et al., 2010).

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques:

Journal: PloS one

Article Title: Aberrant expression of interleukin-1β and inflammasome activation in human malignant gliomas.

doi: 10.1371/journal.pone.0103432

Figure Lengend Snippet: Figure 6. miR-132, miR-212 or the proteasome inhibitor lactacystin do not affect the expression of IL-1 in human astrocytes. Human astrocytes were transfected with specific or control anti-miR inhibitors (10 nM) for 48 h, and then stimulated with IL-1a for 24 h. (A) The expression of miR-132 was quantified by TaqMan real-time RT-PCR. Specific anti-miRs but not control anti-miR suppress miR-132 expression. (B) The culture supernatants were examined for the presence of IL-1b protein by sensitive ELISA with a lower detection limit of 3.9 pg/ml. There was no detectable IL-1b protein production in any of the human astrocyte cultures examined. (C) The effect of the proteasome inhibitor lactacystin on astrocyte IL-1b was examined. Astrocytes were treated with lactacystin at indicated concentrations with or without IL-1a, then cell lysates were subjected to ELISA after 24 h. IL-1b protein was undetectable under any conditions. Mean 6 SD from triplicate cultures. doi:10.1371/journal.pone.0103432.g006

Article Snippet: Lactacystin was purchased from Santa Cruz Biotechnology and was added to culture 10 min prior to cell stimulation.

Techniques: Expressing, Transfection, Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Microbiology Spectrum

Article Title: Condition-dependent effects of Elexacaftor/Tezacaftor/Ivacaftor (Trikafta) on Aspergillus fumigatus growth

doi: 10.1128/spectrum.02275-24

Figure Lengend Snippet: High concentration of CFTR modulators (Elexacaftor [VX445], Tezacaftor [VX661], and Ivacaftor [VX770]) impairs macrophage control of A. fumigatus growth. ( A ) Measurement of A. fumigatus fungal length (Strain: DAL-DSred) infecting bone marrow-derived macrophages (BMDM) at a multiplicity of infection (MOI) of 5, using the IncuCyte live-cell analysis system in the presence of 3VX (VX445/VX661/VX770) treatment at [3VX] S , [3VX] E , or [3VX] 10μM , as well as with [Veh]. * P < 0.05, ** P < 0.01 (two-way ANOVA with Dunnett’s multiple comparisons test); data are presented as mean ± SEM, n = 3. ( B ) Release of TNF⍺ in BMDMs was assessed after 20 h with A. fumigatus (MOI of 15). ** P < 0.01 (one-way ANOVA with Dunnett’s multiple comparisons test); data are presented as mean ± SEM, n = 4. ( C ) Normalized phagocytosis to vehicle control ([Veh]) at 20 h time point. **P < 0.01 , ****P < 0.005 (one sample t test, theoretical mean of 1); data are presented as mean ± SEM, n = 4. ( D ) Representative measurement of zymosan phagocytosis by primed BMDMs in the presence of 3VX treatment at concentrations [3VX] S , [3VX] E or [3VX] 10μM , as well as in the presence of cytochalasine D or [Veh]. ( E ) Quantification of viable conidia following 4 h of BMDM infection in the presence of 3VX treatment at [3VX] S , [3VX] E , or [3VX] 10μM , as well as with [Veh]. ( F ) Immunoblot analysis of pro-caspase-1 ( P45 ), the caspase-1 subunit p20 ( P20 ), and β-actin of primed BMDMs left untreated (medium alone [Med]) or in the presence of 3VX treatment at [3VX] S , [3VX] E , or [3VX] 10μM , as well as with [Veh] during 20 h after infection with A. fumigatus (MOI of 15). ( G ) Release of IL-1β in BMDMs assessed after 20 h infection with A. fumigatus (MOI of 15) left untreated (medium alone [Med]) or in the presence of 3VX treatment at [3VX] S , [3VX] E , or [3VX] 10μM , as well as with [Veh]. *P < 0.05 , **P < 0.01 (one-way ANOVA with Dunnett’s multiple comparisons test); data are presented as mean ± SEM, n = 3.

Article Snippet: Cytokine levels were determined by ELISA using kits according to the manufacturer’s instructions: TNF⍺ DuoSet ELISA (DY410, R&D Systems) and IL-1β (DY401, R&D Systems).

Techniques: Concentration Assay, Control, Derivative Assay, Infection, Cell Analysis, Western Blot

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 primary mouse hepatocytes were treated with PA (S3794, Selleck, USA; 0.5 mmol/L for 24 h), SDF‐1 neutralising antibody (1 μg for 24 h), AMD3100 (CXCR4 antagonist; HY‐10046, MedChemExpress, USA; 10 μM for 24 h), ACT‐1004‐1239 (CXCR7 antagonist; HY‐142617, MedChemExpress; 6 nM for 24 h), MK‐2206 2HCl (AKT inhibitor; HY‐10358, MedChemExpress, 10 μM for 24 h), XL388 (mTOR inhibitor; HY‐13806, MedChemExpress, 100 nM for 24 h), SC79 (AKT activator; HY‐18749, MedChemExpress, 4 μg/mL for 24 h), MHY1485 (mTOR activator; HY‐B0795, MedChemExpress, 5 μM for 24 h), or 3‐MA (10 mM for 24 h).

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 primary mouse hepatocytes were treated with PA (S3794, Selleck, USA; 0.5 mmol/L for 24 h), SDF‐1 neutralising antibody (1 μg for 24 h), AMD3100 (CXCR4 antagonist; HY‐10046, MedChemExpress, USA; 10 μM for 24 h), ACT‐1004‐1239 (CXCR7 antagonist; HY‐142617, MedChemExpress; 6 nM for 24 h), MK‐2206 2HCl (AKT inhibitor; HY‐10358, MedChemExpress, 10 μM for 24 h), XL388 (mTOR inhibitor; HY‐13806, MedChemExpress, 100 nM for 24 h), SC79 (AKT activator; HY‐18749, MedChemExpress, 4 μg/mL for 24 h), MHY1485 (mTOR activator; HY‐B0795, MedChemExpress, 5 μM for 24 h), or 3‐MA (10 mM for 24 h).

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: CXCR4 and CXCR7 expression increase on PA‐treated hepatocytes. The hepatocytes were divided into normal and PA (0.5 mmol/L for 24 h) groups. (A) CXCR4 and CXCR7 protein levels on the plasma membrane of hepatocytes were detected by Western blot. (B, C) CXCR4 and CXCR7 relative protein levels were analysed. ** p < 0.01 versus normal group.

Article Snippet: The primary mouse hepatocytes were treated with PA (S3794, Selleck, USA; 0.5 mmol/L for 24 h), SDF‐1 neutralising antibody (1 μg for 24 h), AMD3100 (CXCR4 antagonist; HY‐10046, MedChemExpress, USA; 10 μM for 24 h), ACT‐1004‐1239 (CXCR7 antagonist; HY‐142617, MedChemExpress; 6 nM for 24 h), MK‐2206 2HCl (AKT inhibitor; HY‐10358, MedChemExpress, 10 μM for 24 h), XL388 (mTOR inhibitor; HY‐13806, MedChemExpress, 100 nM for 24 h), SC79 (AKT activator; HY‐18749, MedChemExpress, 4 μg/mL for 24 h), MHY1485 (mTOR activator; HY‐B0795, MedChemExpress, 5 μM for 24 h), or 3‐MA (10 mM for 24 h).

Techniques: Expressing, Clinical Proteomics, Membrane, Western Blot

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 binds to CXCR4 and CXCR7 on PA‐treated hepatocytes. The hepatocytes were divided into normal and PA (0.5 mmol/L for 24 h) groups. IP was performed to detect the interaction between SDF‐1 and CXCR4 on the plasma membrane of hepatocytes using SDF‐1 (A) or CXCR4 (B) antibody. IP was performed to detect the interaction between SDF‐1 and CXCR7 on the plasma membrane of hepatocytes using SDF‐1 (C) or CXCR7 (D) antibody.

Article Snippet: The primary mouse hepatocytes were treated with PA (S3794, Selleck, USA; 0.5 mmol/L for 24 h), SDF‐1 neutralising antibody (1 μg for 24 h), AMD3100 (CXCR4 antagonist; HY‐10046, MedChemExpress, USA; 10 μM for 24 h), ACT‐1004‐1239 (CXCR7 antagonist; HY‐142617, MedChemExpress; 6 nM for 24 h), MK‐2206 2HCl (AKT inhibitor; HY‐10358, MedChemExpress, 10 μM for 24 h), XL388 (mTOR inhibitor; HY‐13806, MedChemExpress, 100 nM for 24 h), SC79 (AKT activator; HY‐18749, MedChemExpress, 4 μg/mL for 24 h), MHY1485 (mTOR activator; HY‐B0795, MedChemExpress, 5 μM for 24 h), or 3‐MA (10 mM for 24 h).

Techniques: Clinical Proteomics, Membrane

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 primary mouse hepatocytes were treated with PA (S3794, Selleck, USA; 0.5 mmol/L for 24 h), SDF‐1 neutralising antibody (1 μg for 24 h), AMD3100 (CXCR4 antagonist; HY‐10046, MedChemExpress, USA; 10 μM for 24 h), ACT‐1004‐1239 (CXCR7 antagonist; HY‐142617, MedChemExpress; 6 nM for 24 h), MK‐2206 2HCl (AKT inhibitor; HY‐10358, MedChemExpress, 10 μM for 24 h), XL388 (mTOR inhibitor; HY‐13806, MedChemExpress, 100 nM for 24 h), SC79 (AKT activator; HY‐18749, MedChemExpress, 4 μg/mL for 24 h), MHY1485 (mTOR activator; HY‐B0795, MedChemExpress, 5 μM for 24 h), or 3‐MA (10 mM for 24 h).

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 primary mouse hepatocytes were treated with PA (S3794, Selleck, USA; 0.5 mmol/L for 24 h), SDF‐1 neutralising antibody (1 μg for 24 h), AMD3100 (CXCR4 antagonist; HY‐10046, MedChemExpress, USA; 10 μM for 24 h), ACT‐1004‐1239 (CXCR7 antagonist; HY‐142617, MedChemExpress; 6 nM for 24 h), MK‐2206 2HCl (AKT inhibitor; HY‐10358, MedChemExpress, 10 μM for 24 h), XL388 (mTOR inhibitor; HY‐13806, MedChemExpress, 100 nM for 24 h), SC79 (AKT activator; HY‐18749, MedChemExpress, 4 μg/mL for 24 h), MHY1485 (mTOR activator; HY‐B0795, MedChemExpress, 5 μM for 24 h), or 3‐MA (10 mM for 24 h).

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 primary mouse hepatocytes were treated with PA (S3794, Selleck, USA; 0.5 mmol/L for 24 h), SDF‐1 neutralising antibody (1 μg for 24 h), AMD3100 (CXCR4 antagonist; HY‐10046, MedChemExpress, USA; 10 μM for 24 h), ACT‐1004‐1239 (CXCR7 antagonist; HY‐142617, MedChemExpress; 6 nM for 24 h), MK‐2206 2HCl (AKT inhibitor; HY‐10358, MedChemExpress, 10 μM for 24 h), XL388 (mTOR inhibitor; HY‐13806, MedChemExpress, 100 nM for 24 h), SC79 (AKT activator; HY‐18749, MedChemExpress, 4 μg/mL for 24 h), MHY1485 (mTOR activator; HY‐B0795, MedChemExpress, 5 μM for 24 h), or 3‐MA (10 mM for 24 h).

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 primary mouse hepatocytes were treated with PA (S3794, Selleck, USA; 0.5 mmol/L for 24 h), SDF‐1 neutralising antibody (1 μg for 24 h), AMD3100 (CXCR4 antagonist; HY‐10046, MedChemExpress, USA; 10 μM for 24 h), ACT‐1004‐1239 (CXCR7 antagonist; HY‐142617, MedChemExpress; 6 nM for 24 h), MK‐2206 2HCl (AKT inhibitor; HY‐10358, MedChemExpress, 10 μM for 24 h), XL388 (mTOR inhibitor; HY‐13806, MedChemExpress, 100 nM for 24 h), SC79 (AKT activator; HY‐18749, MedChemExpress, 4 μg/mL for 24 h), MHY1485 (mTOR activator; HY‐B0795, MedChemExpress, 5 μM for 24 h), or 3‐MA (10 mM for 24 h).

Techniques: Phospho-proteomics