Journal: Scientific Reports

Article Title: DDR2 ameliorates nonalcoholic hepatic steatosis by activating the AMPK/ACC pathway

doi: 10.1038/s41598-026-42992-0

Figure Lengend Snippet: DDR2 overexpression in hepatocytes mitigates gluconeogenesis and lipid deposition in db/db mice. ( A ) Pattern diagram of treated mice ( n = 6). ( B–C ) DDR2 mRNA ( B ) and protein ( C ) levels in the livers of db/db mice, 11 days following infection with Ad-GFP or Ad-DDR2 ( n = 6). ( D ) Representative electron microscopy images of liver sections, scale bar 1 μm; H&E-stained sections, scale bar 50 μm; Oil Red O-stained sections, scale bar 50 μm and F4/80 staining, scale bar 30 μm. ( E ) Serum glucose concentrations in 6-hour-fasted db/db mice. ( F ) Reduced serum TG induced by DDR2 overexpression. ( G ) GTT performed in Ad-GFP- or Ad-DDR2-injected db/db mice, 9 days post-injection. ( H ) PEPCK and Pgc-1α protein level in liver tissues from the two groups. ( I ) There was no significant change in TG content in the liver. ( J ) AMPK and ACC phosphorylation and Srebp-1c, Fasn and Scd-1 protein levels in liver tissue. ( K ) Reduced serum TNF-α contents following DDR2 overexpression. ( L ) eIF-2α and IRE-1α phosphorylation in mouse livers. Data are shown as mean ± SEM, with comparisons by univariate t-tests. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Serum TNF-α levels were ascertained through an ELISA kit (E-MSEL-M0002, Elabscience Biotechnology Inc., Wuhan, China).

Techniques: Over Expression, Infection, Electron Microscopy, Staining, Injection, Phospho-proteomics

Journal: Advanced Science

Article Title: Neuronal Cholesterol Deficiency Mediated by Astrocytic SREBP2 Downregulation Leads to Postoperative Cognitive Dysfunction Through Impairment of Hippocampal Synaptic Plasticity and Excitatory Synaptic Transmission

doi: 10.1002/advs.202519874

Figure Lengend Snippet: Chemogenetic inhibition of astrocytes in the dCA1 rescued cholesterol synthesis in elderly mice that underwent anesthesia/surgery. (A) Representative immunofluorescence images of hM4D(Gi)‐mCherry (red) and the astrocyte marker GFAP (green) (scale bar, 200 µm). (B) Representative immunofluorescence images of hM4D(Gi)‐mCherry (red) and the neuronal marker NeuN (green) (scale bar, 200 µm). (C) Representative immunofluorescence images of hM4D(Gi)‐mCherry (red) and the microglial marker Iba‐1 (green) (scale bar, 200 µm). (D) hM4D(Gi)‐mCherry was mainly expressed in astrocytes with high penetrance (>96% of the GFAP cells expressed mCherry) ( n = 10 mice/group). (E) hM4D(Gi)‐mCherry was mainly expressed in astrocytes with high specificity (>95% mCherry positive cells were also GFAP‐positive). No co‐localization of hM4D(Gi)‐mCherry with NeuN or Iba1 was detected ( n = 10 mice/group). (F) Representative immunofluorescence images of C3 (green) and GFAP (gray); cells were counterstained with DAPI (blue); the orange arrow indicates the double‐positive cells (40× magnification; scale bar, 50 µm). (G) Density of C3/GFAP co‐labeled cells in the dCA1 ( n = 6 mice/group). Equal variances were confirmed using the F‐test [F(5,5) = 1.684, p = 0.5812]. The data followed a normal distribution; p ‐values were calculated using the unpaired t ‐test. (H) Analysis of IL‐6 levels in the dCA1 ( n = 6 mice/group). Equal variances were confirmed by F‐test [F(5,5) = 1.294, p = 0.784]. The data were normally distributed; p ‐values were calculated using the unpaired t ‐test. (I) Analysis of IL‐1β levels in the dCA1 ( n = 6 mice/group). Equal variances were confirmed using the F‐test [F(5,5) = 1.668, p = 0.588]. The data were normally distributed; p ‐values were calculated using the unpaired t ‐test. (J) Analysis of TNF‐α levels in the dCA1 ( n = 6 mice/group). Equal variances were confirmed using the F‐test [F(5,5) = 4.579, p = 0.1204]. The data were normally distributed; p ‐values were calculated using the unpaired t ‐test. (K) Comparison of SREBP2 protein expression between the A/S+hM4D(Gi)/Saline and A/S+hM4D(Gi)/CNO groups ( n = 6 mice/group). Equal variances were confirmed using the F‐test [F(5,5) = 1.308, p = 0.7757]. The data were normally distributed; p ‐values were calculated using the unpaired t ‐test. (L) Representative immunofluorescence images of SREBP2 (green) and GFAP (gray); cells were counterstained with DAPI (blue) (20× magnification; scale bar, 100 µm). (M) Density of SREBP2/GFAP co‐labeled cells in the dCA1 ( n = 6 mice/group). Equal variances were confirmed using the F‐test [F(5,5) = 2.899, p = 0.2677]. The data were normally distributed; p ‐values were calculated using the unpaired t ‐test. (N) Co‐labeling rate of SREBP2 + /GFAP + cells in the dCA1 ( n = 6 mice/group). Equal variances were confirmed using the F‐test [F(5,5) = 2.183, p = 0.4116]. The data were normally distributed; p ‐values were calculated using the unpaired t ‐test. (O) Representative immunofluorescence images of Filipin (green) (60× magnification; scale bar, 100 µm). (P) Density of Filipin‐positive neurons in the dCA1 ( n = 3 mice/group). Equal variances were confirmed using the F‐test [F(2,2) = 1]. The data were normally distributed; p ‐values were calculated using the unpaired t ‐test. (Q) Cholesterol concentrations in the hippocampus ( n = 6 mice/group). Equal variances were confirmed using the F‐test [F(5,5) = 1.102, p = 0.918]. The data were normally distributed; p ‐values were calculated using the unpaired t ‐test. All data are presented as mean ± SD values. * p < 0.05, ** p < 0.01 vs the A/S+hM4D(Gi)/Saline group. CA1: Cornu Ammonis 1; CA3: Cornu Ammonis 3; DG: Dentate Gyrus.

Article Snippet: Supernatants were collected for analysis, the TNF‐α level was measured using ELISA kit (ABclonal, Cat.: RK00027); the IL‐6 level was measured using ELISA kit (ABclonal, Cat.: RK00008); and the IL‐1β level was measured using ELISA kit (ABclonal, Cat.: RK00006).

Techniques: Inhibition, Immunofluorescence, Marker, Labeling, Comparison, Expressing, Saline

Journal: Advanced Science

Article Title: Neuronal Cholesterol Deficiency Mediated by Astrocytic SREBP2 Downregulation Leads to Postoperative Cognitive Dysfunction Through Impairment of Hippocampal Synaptic Plasticity and Excitatory Synaptic Transmission

doi: 10.1002/advs.202519874

Figure Lengend Snippet: Anesthesia/surgery impaired cognitive function, increased pro‐inflammatory factor levels, and suppressed astrocytic SREBP2 levels in aged mice. (A–C) Hippocampal levels of IL‐6, IL‐1β, and TNF‐α 3 days after surgery ( n = 6 mice/group). Equal variances were confirmed using the Brown–Forsythe test; IL‐6: [F(2, 15) = 0.66, p = 0.5312]; IL‐1β: [F(2, 15) = 0.02894, p = 0.9715]; TNF‐α: [F(2, 15) = 0.4066, p = 0.673]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. (D) Representative confocal images staining for C3 (green) and GFAP (red) in the dCA1. Cells were counterstained with DAPI (blue); the white arrow indicates the double‐positive cells (40× magnification; scale bar, 50 µm). (E) Density of C3/GFAP co‐labeled cells ( n = 6 mice/group). Equal variances were confirmed using the Brown–Forsythe test [F(2, 15) = 0.8925, p = 0.4303]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. (F) Comparison of SREBP2 protein expression among three groups ( n = 6 mice/group). Equal variances were confirmed by Brown‐Forsythe test [F(2, 15) = 0.981, p = 0.3978]. The data follow a normal distribution, p ‐values are calculated using one‐way ANOVA with Tukey's post hoc test. (G) Representative immunofluorescence images of SREBP2 (green) and GFAP (red) in the dCA1; cells were counterstained with DAPI (blue) (20× magnification; scale bar, 100 µm). (H) Density of SREBP2/GFAP co‐labeled cells ( n = 6 mice/group). Equal variances were confirmed using the Brown–Forsythe test [F(2, 15) = 0.557, p = 0.5843]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. (I) Co‐labeling rate of SREBP2 + /GFAP + cells ( n = 6 mice per group). Equal variances were confirmed using the Brown–Forsythe test [F(2, 15) = 0.6318, p = 0.5452]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. (J) Representative immunofluorescence images of Filipin (green) in the dCA1 (60× magnification; scale bar, 100 µm). (K) Density of Filipin‐positive neurons in dCA1 ( n = 4 mice/group). Brown–Forsythe test [F(2, 9) = 1, p = 0.4053]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. (L) Cholesterol concentration in the hippocampus ( n = 6 mice/group). Brown‐Forsythe test [F(2, 15) = 0.9641, p = 0.4038]. The data follow a normal distribution, p ‐values are calculated using one‐way ANOVA with Tukey's post hoc test. (M) Representative trajectory in the Y‐maze test. (N) Comparison of total moving distances among the three groups ( n = 13 mice per group). Brown–Forsythe test [F(2, 36) = 2.069, p = 0.141]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. (O) Comparison of the total moving distance in the novel arm among the three groups ( n = 13 mice/group). Brown–Forsythe test [F(2, 36) = 2.305, p = 0.1143]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. (P) Comparison of time spent in the novel arm among the three groups ( n = 13 mice per group). Brown–Forsythe test [F(2, 36) = 1.213, p = 0.3092]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. (Q) Representative NORT trajectory. (R) Comparison of total object exploration time among the three groups ( n = 13 mice per group). Brown–Forsythe test [F(2, 36) = 1.547, p = 0.2268]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. (S) Percentage of time spent exploring a novel object ( n = 13 mice per group). Brown–Forsythe test [F(2, 36) = 1.965, p = 0.1549]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. (T) Representative trajectory in the OLT. (U) Comparison of the total object exploration time among the three groups ( n = 13 mice per group). Brown–Forsythe test [F(2, 36) = 0.03795, p = 0.9628]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. (V) Percentage of time spent exploring a novel object ( n = 13 mice per group). Brown–Forsythe test [F(2, 36) = 2.405, p = 0.1046]. The data were normally distributed; p ‐values were calculated using one‐way ANOVA with Tukey's post hoc test. All data are presented as mean ± SD values. ** p < 0.01 vs the Saline group; # p < 0.05, ## p < 0.01 vs the A/S+Saline group.

Article Snippet: Supernatants were collected for analysis, the TNF‐α level was measured using ELISA kit (ABclonal, Cat.: RK00027); the IL‐6 level was measured using ELISA kit (ABclonal, Cat.: RK00008); and the IL‐1β level was measured using ELISA kit (ABclonal, Cat.: RK00006).

Techniques: Staining, Labeling, Comparison, Expressing, Immunofluorescence, Concentration Assay, Saline

Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: rhIL-37 delays preterm delivery by reducing LPS-induced inflammatory effects and NLRP3 inflammasome activation in mice. ( A ) Orbital blood was collected from the control mice ( n = 6), LPS-induced preterm mice (LPS; n = 6), and rhIL-37 + LPS-co-treated preterm mice (rhIL-37 + LPS; n = 6).Serum levels of IL-1β, IL-6 and TNF-α were detected via ELISA. ( B ) Foetal membrane and placental tissues were collected from mice ( n = 6 per group) immediately before labour,, and the mRNA expression levels of NLRP3, Caspase-1 and ASC were detected via qRT-PCR. ( C ) Foetal membrane and placental tissues were collected from the mice in each group ( n = 3 per group), and the protein levels of NLRP3, Caspase-1, Pro-caspase-1 and ASC were analyzed by western blotting. ( D ) Western blot Results were normalised to β-actin. Bar graphs show statistical quantification of the western blot data. All data met the assumption of homogeneity of variance. Data are presented as mean ± s.e.m. Results are representative of at least three independent experiments. * p < 0.05; ** p < 0.01;*** p < 0.001;**** p < 0.0001 (one-way ANOVA).

Article Snippet: Supernatants collected from HTR-8/SVneo cells were assayed for IL-1β, IL-6, and TNF-α levels using a human ELISA kit (LiankeBio, Hangzhou, China).

Techniques: Activation Assay, Control, Enzyme-linked Immunosorbent Assay, Membrane, Expressing, Quantitative RT-PCR, Western Blot

Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: IL-37 knockdown enhances LPS-induced inflammatory and NLRP3 inflammasome activation in HTR-8/Svneo cells. ( A ) Cell supernatants were collected from control, si_NC- or si_IL37-treated cells following LPS exposure, and IL-37 expression level were detected by ELISA, qRT-PCR and Western blotting. ( B ) IL-1β, IL-6 and TNF-α levels in the cell supernatants were detected by ELISA. ( C ) The mRNA expression levels of NLRP3, Caspase-1 and ASC after cell treatment with LPS ( n = 5) were measured by qRT-PCR analysis. ( D ) NLRP3,pro-caspase-1, caspase-1 and ASC protein levels were detected by Western blotting, and the results were normalised to β-actin. All data exhibited equal variance.The data are shown as the mean ± s.e.m. The results are representative of at least three independent experiments. * p < 0.05; ** p < 0.01;*** p < 0.001;**** p < 0.0001 (One-way ANOVA).

Article Snippet: Supernatants collected from HTR-8/SVneo cells were assayed for IL-1β, IL-6, and TNF-α levels using a human ELISA kit (LiankeBio, Hangzhou, China).

Techniques: Knockdown, Activation Assay, Control, Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: Nigericin reversed the anti-inflammatory effects and effects of rhIL-37 and its inhibition of NF-κB p65 activation in LPS-induced PTB.t ( A ) Cell supernatants were collected from HTR-8/Svneo cells co-treated with rhIL-37 and LPS, then either left untreated or treated with nigericin.The mRNA expression of NLRP3, caspase-1, and ASC was detected. ( B ) Cells were collected, and the protein expression of NLRP3, caspase-1, pro-caspase-1, and ASC was detected by western blotting. Results were normalised to β-actin and statistically analyzed.( C ) HTR-8/Svneo cell supernatants were collected after co-treatment with rhIL-37 and LPS (with or without nigericin), and the expression levels of IL-1β, IL-6, and TNF-α were detected by ELISA. ( D ) HTR-8/Svneo cells co-treated with rhIL-37 and LPS (with or without nigericin) were collected. The mRNA expression of p65 was detected by qRT-PCR, and protein expression of p65 and p-p65 was detected by western blotting. Densitometric quantification of p-p65 relative to p65 was performed. Statistical analysis of the western blot results was performed.All data exhibited equal variance.The data are shown as the means ± s.e.m. The results are representative of at least three independent experiments..

Article Snippet: Supernatants collected from HTR-8/SVneo cells were assayed for IL-1β, IL-6, and TNF-α levels using a human ELISA kit (LiankeBio, Hangzhou, China).

Techniques: Inhibition, Activation Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR