Journal: eLife

Article Title: Oxidized low-density lipoprotein potentiates angiotensin II-induced Gq activation through the AT1-LOX1 receptor complex

doi: 10.7554/eLife.98766

Figure Lengend Snippet: ( a ) A Schematic protocol for the animal experiments. Eight-wk-old male wild-type (WT) mice and male LOX-1 KO mice were fed either an normal diet (ND) or an high-fat diet (HFD) for 6 wk. After 10 wk of age, the mice were treated over a 4 wk period with infusions of either saline or Ang II. Ang II was administered at two dosage levels: a subpressor dose of 0.1 γ and a pressor dose of 0.7 γ, delivered via subcutaneously implanted osmotic pumps. At the end of the infusion period, urine was collected, the animals were sacrificed, and comprehensive tissue analysis was conducted to evaluate the renal effects of the treatments. ( b ) Average systolic blood pressure (SBP) measured at half-week intervals in WT and LOX-1 KO mice during the 4 wk infusion period. ( c, d ) Urine 8-OHDG concentrations (mg/g creatinine [Cr]) ( c ) and urine albumin concentrations (mg/g creatinine [Cr]) ( d ) in WT and LOX-1 KO mice at the conclusion of the 4 wk infusion period. Data are represented as mean ± SEM. Differences were determined by one-way ANOVA, followed by Tukey’s multiple comparison test ( a-d ). Figure 7—source data 1. Oxidized low-density lipoprotein (LDL) inducible diet exacerbates angiotensin II (Ang II)-induced renal dysfuntion in wild-type mice, but not in LOX-1 knockout mice. Source data for .

Article Snippet: Mice were housed in a temperature-controlled (20–22°C) room on a 12 hr light/dark cycle and fed an ND (MF; Oriental Yeast, Osaka, Japan) or an HFD (High Fat Diet without DL-α-tocopherol, CLEA Japan Inc, Tokyo, Japan), which reported to increase plasma LOX-1 ligand in ApoE KO mice ( ).

Techniques: Saline, Comparison, Knock-Out

Journal: eLife

Article Title: Oxidized low-density lipoprotein potentiates angiotensin II-induced Gq activation through the AT1-LOX1 receptor complex

doi: 10.7554/eLife.98766

Figure Lengend Snippet: Plasma LOX-1 ligand concentration of 14-wk-old wild-type mice upon normal diet (ND) and high-fat diet (HFD) for 6 wk. From 10 wk of age, these mice also received concurrent 4 wk infusions of angiotensin II (Ang II) at a pressor dose of 0.7 γ, administered through subcutaneously implanted osmotic pumps. Data are represented as mean ± SEM. Differences were determined using Student’s t-test (n=7 for each group). Figure 7—figure supplement 1—source data 1. High-fat diet used in the study prominently increased plasma LOX-1 ligand concentration. Source data for .

Article Snippet: Mice were housed in a temperature-controlled (20–22°C) room on a 12 hr light/dark cycle and fed an ND (MF; Oriental Yeast, Osaka, Japan) or an HFD (High Fat Diet without DL-α-tocopherol, CLEA Japan Inc, Tokyo, Japan), which reported to increase plasma LOX-1 ligand in ApoE KO mice ( ).

Techniques: Clinical Proteomics, Concentration Assay

Journal: eLife

Article Title: Oxidized low-density lipoprotein potentiates angiotensin II-induced Gq activation through the AT1-LOX1 receptor complex

doi: 10.7554/eLife.98766

Figure Lengend Snippet: ( a ) Final Body weight: This figure shows the body weights of wild-type (WT) and LOX-1 KO mice at the end of the 4 wk infusion period, highlighting the effects of diet and pharmacological treatment. ( b, c ) Serial Body Weight Changes: These graphs depict the progression of body weight over time in WT and LOX-1 KO mice, illustrating the impact of the dietary regimen and Ang II infusion on weight dynamics. ( d, e ) Food Intake: Food intake Trajectory: Serial measurements of weekly food intake (g/wk per mouse) are presented for WT and LOX-1 KO mice. The Figures show the changes in food intake over the course of the study, corresponding to the administration of a pressor dose of 0.7 γ (d) and a subpressor dose of 0.1 γ (e) of Ang II, respectively. ( f, g ) Systolic Blood Pressure Trajectory: Serial measurements of systolic blood pressure (SBP) obtained using the tail-cuff method are presented for WT and LOX-1 KO mice. The Figures show the changes in SBP over the course of the study, corresponding to the administration of a pressor dose of 0.7 γ (d) and a subpressor dose of 0.1 γ ( e ) of Ang II, respectively. Beginning at 8 wk of age, mice were fed either an normal diet (ND) or an high-fat diet (HFD) for 6 wk. From 10 wk of age, coinciding with the 2 wk time point in the Figure, the mice underwent a 4 wk period of infusion with either vehicle or Ang II. The infusion was delivered at specific dosage levels through subcutaneously implanted osmotic pumps. Data are represented as mean ± SEM. Differences were determined using one-way ANOVA, followed by Tukey’s multiple comparison test for ( a ). Figure 7—figure supplement 2—source data 1. Impact of diet and angiotensin II (Ang II) infusion on body weight and systolic blood pressure in mice. Source data for .

Article Snippet: Mice were housed in a temperature-controlled (20–22°C) room on a 12 hr light/dark cycle and fed an ND (MF; Oriental Yeast, Osaka, Japan) or an HFD (High Fat Diet without DL-α-tocopherol, CLEA Japan Inc, Tokyo, Japan), which reported to increase plasma LOX-1 ligand in ApoE KO mice ( ).

Techniques: Comparison

Journal: eLife

Article Title: Oxidized low-density lipoprotein potentiates angiotensin II-induced Gq activation through the AT1-LOX1 receptor complex

doi: 10.7554/eLife.98766

Figure Lengend Snippet: Plasma aldosterone concentration in 14-wk-old wild-type mice fed an normal diet (ND) or an high-fat diet (HFD) for 6 wk. From 10 wk of age, these mice also received concurrent 4 wk infusions of Ang II at a pressor dose of 0.7 γ, administered through subcutaneously implanted osmotic pumps. Data are represented as mean ± SEM. Differences were determined using Student’s t-test (n=7 for each group). Figure 7—figure supplement 3—source data 1. No significant difference was found in plasma aldosterone concentration between a normal diet and a high fat diet-fed wild-type mice with a pressor dose of angiotensin II (Ang II). Source data for .

Article Snippet: Mice were housed in a temperature-controlled (20–22°C) room on a 12 hr light/dark cycle and fed an ND (MF; Oriental Yeast, Osaka, Japan) or an HFD (High Fat Diet without DL-α-tocopherol, CLEA Japan Inc, Tokyo, Japan), which reported to increase plasma LOX-1 ligand in ApoE KO mice ( ).

Techniques: Clinical Proteomics, Concentration Assay

Journal: eLife

Article Title: Oxidized low-density lipoprotein potentiates angiotensin II-induced Gq activation through the AT1-LOX1 receptor complex

doi: 10.7554/eLife.98766

Figure Lengend Snippet: Quantitative real-time PCR analysis for gene expression of NADPH components ( Ncf2 and Cybb ), inflammatory cytokines ( Il6, Tnf, Il1b, and Ccl2 ), and fibrosis markers ( Tgfb2, Fn1, Col1a1, and Col4a1 ) in the kidney harvested from wild-type (WT) and LOX-1 KO mice. The experimental procedures, including the dietary regimen and Ang II administration, are detailed in . Mice were administered either a pressor dose of 0.7 γ Ang II ( a ) or a subpressor dose of 0.1 γ Ang II ( b ). Data are represented as mean ± SEM. Differences were determined using one-way ANOVA, followed by Tukey’s multiple comparison test for ( a ) and ( b ). Figure 8—source data 1. A high-fat diet enhanced angiotensin II (Ang II)-induced renal injury-related gene expression in the kidney in a LOX-1-dependent manner. Source data for .

Article Snippet: Mice were housed in a temperature-controlled (20–22°C) room on a 12 hr light/dark cycle and fed an ND (MF; Oriental Yeast, Osaka, Japan) or an HFD (High Fat Diet without DL-α-tocopherol, CLEA Japan Inc, Tokyo, Japan), which reported to increase plasma LOX-1 ligand in ApoE KO mice ( ).

Techniques: Real-time Polymerase Chain Reaction, Gene Expression, Comparison

Journal: eLife

Article Title: Oxidized low-density lipoprotein potentiates angiotensin II-induced Gq activation through the AT1-LOX1 receptor complex

doi: 10.7554/eLife.98766

Figure Lengend Snippet: Quantitative real-time PCR analysis for gene expression of NADPH components ( Ncf4 and Ncf1 ), inflammatory gene ( Ptgs2 ), fibrosis markers ( Acta2 and Vim ), epithelial markers ( Cdh1 and Chd16 ), tubular marker ( Ptgs2 ), Agtr1a , Agtr1b , and Olr1 in the kidney harvested from wild-type (WT) and LOX-1 KO mice. The experimental procedures, including the dietary regimen and Ang II administration, are detailed in . Mice were administered either a pressor dose of 0.7 γ Ang II ( a ) or a subpressor dose of 0.1 γ Ang II ( b ). Data are represented as mean ± SEM. Differences were determined using one-way ANOVA, followed by Tukey’s multiple comparison test for ( a ) and ( b ). Figure 8—figure supplement 1—source data 1. A high-fat diet enhanced angiotensin II (Ang II)-induced renal injury-related gene expression in the kidney in a LOX-1-dependent manner. Source data for .

Article Snippet: Mice were housed in a temperature-controlled (20–22°C) room on a 12 hr light/dark cycle and fed an ND (MF; Oriental Yeast, Osaka, Japan) or an HFD (High Fat Diet without DL-α-tocopherol, CLEA Japan Inc, Tokyo, Japan), which reported to increase plasma LOX-1 ligand in ApoE KO mice ( ).

Techniques: Real-time Polymerase Chain Reaction, Gene Expression, Marker, Comparison

Journal: eLife

Article Title: Oxidized low-density lipoprotein potentiates angiotensin II-induced Gq activation through the AT1-LOX1 receptor complex

doi: 10.7554/eLife.98766

Figure Lengend Snippet: ( a ) Left: Representative histological images of Masson-Trichrome staining used to detect fibrosis in renal tissues harvested from wild-type (WT) and LOX-1 KO mice. Right: Quantitative analysis by Masson-Trichrome staining. Data are represented as mean ± SEM. Differences were determined using one-way ANOVA, followed by Tukey’s multiple comparison test. ( b ) Representative histological images of renal tissues harvested from WT and LOX-1 KO mice stained for periodic acid-Schiff (PAS) to assess mesangial expansion and glomerular area, providing insight into the structural integrity of the glomeruli. Eight-week-old mice were fed an normal diet (ND) or an high-fat diet (HFD) for 6 wk. From 10 wk of age, these mice were concurrently treated for 4 wk with infusions of vehicle or Ang II (a subpressor dose of 0.1 γ or a pressor dose of 0.7 γ) through subcutaneously implanted osmotic pomps. Figure 9—figure supplement 1—source data 1. The treatment with angiotensin II (Ang II), a high fat diet, or their combination for 4 wk did not induce any histological changes indicative of renal injury. Source data for quantitative analysis for Masson-Trichrome staining in .

Article Snippet: Mice were housed in a temperature-controlled (20–22°C) room on a 12 hr light/dark cycle and fed an ND (MF; Oriental Yeast, Osaka, Japan) or an HFD (High Fat Diet without DL-α-tocopherol, CLEA Japan Inc, Tokyo, Japan), which reported to increase plasma LOX-1 ligand in ApoE KO mice ( ).

Techniques: Staining, Comparison