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AA exacerbated podocyte injury in PAN‐induced rats. Eight‐week‐old male Sprague–Dawley rats (180–200 g) were subjected to puromycin aminonucleoside (PAN)‐induced podocyte injury by a single intraperitoneal injection of PAN (7 mg/100 g body weight). Ten days after PAN injection, rats in the PAN+AA group received arachidonic acid (AA, 3 mg/kg, i.p.) to evaluate AA‐associated exacerbation of established PAN‐induced injury. (A) Serum AA levels were measured by <t>ELISA.</t> Serum albumin, serum creatinine, 24‐h urinary albumin, serum cholesterol and serum triglyceride levels were measured as indicators of renal function and metabolic status ( n = 6). (B) Representative haematoxylin and eosin (H&E), Masson's trichrome, and periodic acid‐Schiff (PAS) staining of kidney sections ( n = 6). Scale bar, 20 μm. (C) Representative transmission electron microscopy images of podocyte foot processes and quantitative analysis of glomerular basement membrane (GBM) thickness, foot‐process width, and the number of foot processes per μm GBM ( n = 3). Scale bar, 5.0 μm. (D) Representative immunofluorescence staining of nephrin in glomeruli. Nephrin is shown in red and nuclei are counterstained with DAPI in blue ( n = 6). Scale bar, 20 μm. (E) Representative Western blot analysis and quantification of nephrin protein expression in renal tissue lysates ( n = 6). Data are expressed as the mean ± SD and analysed by one‐way ANOVA with Bonferroni correction. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant. AA, arachidonic acid; GBM, glomerular basement membrane; i.p., intraperitoneal; MCD, minimal change disease; PAN, puromycin aminonucleoside.
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AA exacerbated podocyte injury in PAN‐induced rats. Eight‐week‐old male Sprague–Dawley rats (180–200 g) were subjected to puromycin aminonucleoside (PAN)‐induced podocyte injury by a single intraperitoneal injection of PAN (7 mg/100 g body weight). Ten days after PAN injection, rats in the PAN+AA group received arachidonic acid (AA, 3 mg/kg, i.p.) to evaluate AA‐associated exacerbation of established PAN‐induced injury. (A) Serum AA levels were measured by <t>ELISA.</t> Serum albumin, serum creatinine, 24‐h urinary albumin, serum cholesterol and serum triglyceride levels were measured as indicators of renal function and metabolic status ( n = 6). (B) Representative haematoxylin and eosin (H&E), Masson's trichrome, and periodic acid‐Schiff (PAS) staining of kidney sections ( n = 6). Scale bar, 20 μm. (C) Representative transmission electron microscopy images of podocyte foot processes and quantitative analysis of glomerular basement membrane (GBM) thickness, foot‐process width, and the number of foot processes per μm GBM ( n = 3). Scale bar, 5.0 μm. (D) Representative immunofluorescence staining of nephrin in glomeruli. Nephrin is shown in red and nuclei are counterstained with DAPI in blue ( n = 6). Scale bar, 20 μm. (E) Representative Western blot analysis and quantification of nephrin protein expression in renal tissue lysates ( n = 6). Data are expressed as the mean ± SD and analysed by one‐way ANOVA with Bonferroni correction. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant. AA, arachidonic acid; GBM, glomerular basement membrane; i.p., intraperitoneal; MCD, minimal change disease; PAN, puromycin aminonucleoside.
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AA exacerbated podocyte injury in PAN‐induced rats. Eight‐week‐old male Sprague–Dawley rats (180–200 g) were subjected to puromycin aminonucleoside (PAN)‐induced podocyte injury by a single intraperitoneal injection of PAN (7 mg/100 g body weight). Ten days after PAN injection, rats in the PAN+AA group received arachidonic acid (AA, 3 mg/kg, i.p.) to evaluate AA‐associated exacerbation of established PAN‐induced injury. (A) Serum AA levels were measured by <t>ELISA.</t> Serum albumin, serum creatinine, 24‐h urinary albumin, serum cholesterol and serum triglyceride levels were measured as indicators of renal function and metabolic status ( n = 6). (B) Representative haematoxylin and eosin (H&E), Masson's trichrome, and periodic acid‐Schiff (PAS) staining of kidney sections ( n = 6). Scale bar, 20 μm. (C) Representative transmission electron microscopy images of podocyte foot processes and quantitative analysis of glomerular basement membrane (GBM) thickness, foot‐process width, and the number of foot processes per μm GBM ( n = 3). Scale bar, 5.0 μm. (D) Representative immunofluorescence staining of nephrin in glomeruli. Nephrin is shown in red and nuclei are counterstained with DAPI in blue ( n = 6). Scale bar, 20 μm. (E) Representative Western blot analysis and quantification of nephrin protein expression in renal tissue lysates ( n = 6). Data are expressed as the mean ± SD and analysed by one‐way ANOVA with Bonferroni correction. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant. AA, arachidonic acid; GBM, glomerular basement membrane; i.p., intraperitoneal; MCD, minimal change disease; PAN, puromycin aminonucleoside.
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AA exacerbated podocyte injury in PAN‐induced rats. Eight‐week‐old male Sprague–Dawley rats (180–200 g) were subjected to puromycin aminonucleoside (PAN)‐induced podocyte injury by a single intraperitoneal injection of PAN (7 mg/100 g body weight). Ten days after PAN injection, rats in the PAN+AA group received arachidonic acid (AA, 3 mg/kg, i.p.) to evaluate AA‐associated exacerbation of established PAN‐induced injury. (A) Serum AA levels were measured by ELISA. Serum albumin, serum creatinine, 24‐h urinary albumin, serum cholesterol and serum triglyceride levels were measured as indicators of renal function and metabolic status ( n = 6). (B) Representative haematoxylin and eosin (H&E), Masson's trichrome, and periodic acid‐Schiff (PAS) staining of kidney sections ( n = 6). Scale bar, 20 μm. (C) Representative transmission electron microscopy images of podocyte foot processes and quantitative analysis of glomerular basement membrane (GBM) thickness, foot‐process width, and the number of foot processes per μm GBM ( n = 3). Scale bar, 5.0 μm. (D) Representative immunofluorescence staining of nephrin in glomeruli. Nephrin is shown in red and nuclei are counterstained with DAPI in blue ( n = 6). Scale bar, 20 μm. (E) Representative Western blot analysis and quantification of nephrin protein expression in renal tissue lysates ( n = 6). Data are expressed as the mean ± SD and analysed by one‐way ANOVA with Bonferroni correction. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant. AA, arachidonic acid; GBM, glomerular basement membrane; i.p., intraperitoneal; MCD, minimal change disease; PAN, puromycin aminonucleoside.

Journal: Journal of Cellular and Molecular Medicine

Article Title: Arachidonic Acid Disrupts PKM2 ‐Linked Glycolysis‐Related Metabolism and Exacerbates PAN ‐Induced Podocyte Injury

doi: 10.1111/jcmm.71404

Figure Lengend Snippet: AA exacerbated podocyte injury in PAN‐induced rats. Eight‐week‐old male Sprague–Dawley rats (180–200 g) were subjected to puromycin aminonucleoside (PAN)‐induced podocyte injury by a single intraperitoneal injection of PAN (7 mg/100 g body weight). Ten days after PAN injection, rats in the PAN+AA group received arachidonic acid (AA, 3 mg/kg, i.p.) to evaluate AA‐associated exacerbation of established PAN‐induced injury. (A) Serum AA levels were measured by ELISA. Serum albumin, serum creatinine, 24‐h urinary albumin, serum cholesterol and serum triglyceride levels were measured as indicators of renal function and metabolic status ( n = 6). (B) Representative haematoxylin and eosin (H&E), Masson's trichrome, and periodic acid‐Schiff (PAS) staining of kidney sections ( n = 6). Scale bar, 20 μm. (C) Representative transmission electron microscopy images of podocyte foot processes and quantitative analysis of glomerular basement membrane (GBM) thickness, foot‐process width, and the number of foot processes per μm GBM ( n = 3). Scale bar, 5.0 μm. (D) Representative immunofluorescence staining of nephrin in glomeruli. Nephrin is shown in red and nuclei are counterstained with DAPI in blue ( n = 6). Scale bar, 20 μm. (E) Representative Western blot analysis and quantification of nephrin protein expression in renal tissue lysates ( n = 6). Data are expressed as the mean ± SD and analysed by one‐way ANOVA with Bonferroni correction. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant. AA, arachidonic acid; GBM, glomerular basement membrane; i.p., intraperitoneal; MCD, minimal change disease; PAN, puromycin aminonucleoside.

Article Snippet: Serum arachidonic acid (AA) levels were measured using a commercially available enzyme‐linked immunosorbent assay (ELISA) kit (Elabscience, catalogue no. E‐EL‐0051) according to the manufacturer's instructions.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Staining, Transmission Assay, Electron Microscopy, Analysis, Membrane, Immunofluorescence, Western Blot, Expressing, Tissue