Journal: BMC Pulmonary Medicine

Article Title: Astragaloside IV restrains pyroptosis and fibrotic development of pulmonary artery smooth muscle cells to ameliorate pulmonary artery hypertension through the PHD2/HIF1α signaling pathway

doi: 10.1186/s12890-023-02660-9

Figure Lengend Snippet: AS-IV reduced pyroptosis and fibrosis development in hypoxia-treated PASMCs. PASMCs were divided into 3 groups: Nor, Hyp and AS, cultured with normoxia or hypoxia, and treated with or without AS-IV. A - D , Cropped blots of GSDMD-N, NLRP3, cleaved Caspase-1, Fibronectin and Collagen1 and quantification of grayscale value (the samples derived from the same experiment and that gels/blots were processed in parallel). E , IL-1β and IL-18 levels in cell supernatant were measured by ELISA. F , Cell death was detected by PI/Hoechst double fluorescent staining. Scale bar = 50 μm. G, LDH activity was detected by a LDH release agent. H - I , Proteins related to extracellular matrix metabolism, including MMP2/9 and TIMP4 were detected by ELISA method. * P <0.05, ** P <0.01, *** P <0.001, comparison with Nor group. # P <0.05, ## P <0.01, ### P <0.001, comparison with Hyp group

Article Snippet: The nuclei were counterstained with Hoechst 33,258 (Abcam, #ab228550), and the image was visualized under a microscope and analyzed by the image J software.

Techniques: Cell Culture, Derivative Assay, Enzyme-linked Immunosorbent Assay, Staining, Activity Assay, Comparison

Journal: BMC Pulmonary Medicine

Article Title: Astragaloside IV restrains pyroptosis and fibrotic development of pulmonary artery smooth muscle cells to ameliorate pulmonary artery hypertension through the PHD2/HIF1α signaling pathway

doi: 10.1186/s12890-023-02660-9

Figure Lengend Snippet: AS-IV depressed pyroptosis and the production of fibrosis markers induced by hypoxia in PASMCs through the PHD2-HIF1α axis. A - D , PASMCs were divided into 5 groups: Nor, Hyp, AS, AS + si-PHD2 and LW6. Cropped blots of GSDMD-N, NLRP3, cleaved Caspase-1, Fibronectin and Collagen1 (the samples derived from the same experiment and that gels/blots were processed in parallel), and quantitation of the pyroptosis markers and the fibrosis markers. E , Cell death was detected by PI/Hoechst double fluorescent staining. Scale bar = 50 μm. F , IL-1β and IL-18 levels in cell supernatant were measured by ELISA. G, LDH activity was detected by a LDH release agent. H - I , Proteins of MMP2/9 and TIMP4 were detected by ELISA method. * P <0.05, ** P <0.01, *** P <0.001, comparison with Nor group. # P <0.05, ## P <0.01, ### P <0.001, comparison with Hyp group. ^ P <0.05, ^^ P <0.01, ^^^ P <0.001, comparison with AS group

Article Snippet: The nuclei were counterstained with Hoechst 33,258 (Abcam, #ab228550), and the image was visualized under a microscope and analyzed by the image J software.

Techniques: Derivative Assay, Quantitation Assay, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, Comparison

Journal: Molecular Medicine

Article Title: Liraglutide attenuates type 2 diabetes mellitus-associated non-alcoholic fatty liver disease by activating AMPK/ACC signaling and inhibiting ferroptosis

doi: 10.1186/s10020-023-00721-7

Figure Lengend Snippet: Liraglutide inhibits sensitivity of high glucose on ferroptosis. A Cell viability in HepG2 cells treated with indicated concentrations of high glucose (HG) for 24 h and 48 h. B Cell viability in Hep3B cells treated with HG (125 mM, 48 h) and combined with 5 µM Z-VAD-FMK (Z-VAD, 24 h), 2 µM Necrostatin-1 s (Nec-1 s, 24 h) and 5 µM ferrostatin-1 (Fer-1, 24 h) . Bar graphs showing cell viability in indicated cells treated with HG (125 mM, 48 h) or erastin (10 µM, 24 h) (C) , RSL3 (1.0 µM, 24 h) D and combined with 5 µM Z-VAD-FMK (Z-VAD, 24 h), 2 µM Necrostatin-1 s (Nec-1 s, 24 h) and 5 µM ferrostatin-1 (Fer-1, 24 h). PLC cells viability were measured with 1 mM, 5 mM, 100 mM glucose (48 h) combined with erastin (24 h) E or RSL3 (24 h) (F) . G Cell viability measured in HepG2 cells treated with 125 mM HG and different concentrations of liraglutide for 48 h. H – I HepG2 and Hep3B cells were treated with 100 mM HG, erastin (5 μM) and combined with 1.0 μM liraglutide for 24 h. J , K HepG2 and Hep3B cells were treated with 100 mM HG, RSL3 (0.5 μM) and combined with 1.0 μM liragluide for 24 h. Cell viability was determined by CCK-8 assays. L Cell viability measured in HepG2 cells treated with different concentrations of erastin (24 h) combined with 2.0 μM liraglutide (48 h). M Cell viability measured in HepG2 cells treated with different concentrations of RSL3 (24 h) combined with 2.0 μM liraglutide (48 h). N Representative images from immunohistochemical staining of liver tissue microarray for Cleaved caspase-3 and 4-HNE (scale bar, 50 μm). O Representative Fe 2+ fluorescence images of HepG2 cell (scale bar, 100 μm) Data are expressed as mean ± SD, n = 4

Article Snippet: Cell viability was measured as described previously using the Cell Counting Kit-8 (CCK8) (Enogene, Nanjing, China).

Techniques: CCK-8 Assay, Immunohistochemistry, Staining, Microarray, Fluorescence