Journal: Comprehensive Physiology

Article Title: Amyloid‐Forming Amylin Secreted by the Pancreas Promotes Hypoxia‐Inducible Factor Activation and Mitochondrial Alterations in Liver and Heart During Type 2 Diabetes Pathogenesis

doi: 10.1002/cph4.70252

Figure Lengend Snippet: Hypoxia Inducible Factor (HIF) accumulation in the livers of human amylin‐expressing HIP rats. (A) Blood glucose concentrations in wild‐type, hyperglycemic (UCD), human amylin‐expressing (HIP), and amylin‐knockout (AKO) rats. Concentrations of (B) HIF‐1α and (C) HIF‐2α measured by ELISA in the livers of WT, UCD, HIP and AKO rats ( n = 6 per group). Expression (relative to wild‐type rats) of HIF target genes (D) Arg1 (encoding arginase 1), (E) Ca9 (encoding carbonic anhydrase 9) and (F) Vegfa (encoding vascular endothelial growth factor a), ( n = 4–5 per group). One‐way ANOVA with post hoc Tukey's multiple comparison test. Data represented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: A competitive ELISA assay kit (EIA‐AMY‐1, Ray‐biotech) was used to measure the amylin content in whole heart homogenates from age‐matched male WT, HIP, and UCD rats.

Techniques: Expressing, Knock-Out, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Comprehensive Physiology

Article Title: Amyloid‐Forming Amylin Secreted by the Pancreas Promotes Hypoxia‐Inducible Factor Activation and Mitochondrial Alterations in Liver and Heart During Type 2 Diabetes Pathogenesis

doi: 10.1002/cph4.70252

Figure Lengend Snippet: Amylin accumulation in the hearts of HIP rats. (A) Representative immunofluorescence images of heart sections from wild‐type (WT), hyperglycemic (UCD), human amylin‐expressing (HIP) rats stained with an anti‐amylin antibody. Scale bar = 20 μm. (B) Amylin levels, measured by ELISA, in heart homogenates from age‐matched WT, UCD and HIP rats. Statistical analysis used the Kruskal–Wallis test ( p = 0.0019) followed by Dunn's post hoc test to compare individual group pairs ( n = 7 per group). Data represented as mean ± SEM.

Article Snippet: A competitive ELISA assay kit (EIA‐AMY‐1, Ray‐biotech) was used to measure the amylin content in whole heart homogenates from age‐matched male WT, HIP, and UCD rats.

Techniques: Immunofluorescence, Expressing, Staining, Enzyme-linked Immunosorbent Assay

Journal: Comprehensive Physiology

Article Title: Amyloid‐Forming Amylin Secreted by the Pancreas Promotes Hypoxia‐Inducible Factor Activation and Mitochondrial Alterations in Liver and Heart During Type 2 Diabetes Pathogenesis

doi: 10.1002/cph4.70252

Figure Lengend Snippet: Hypoxia Inducible Factor (HIF) accumulation in the hearts of human amylin‐expressing HIP rats. (A) Representative immunohistochemistry images of heart sections from wild‐type (WT), hyperglycemic (UCD), human amylin‐expressing (HIP) rats dual‐stained with antibodies against amylin (red/brown signal) and HIF‐2α (green signal). Scale bar—20 μm. (B) HIF‐1α measured by ELISA in the hearts of WT, UCD, and HIP rats and (C) HIF‐2α in the hearts of WT, UCD, HIP, and amylin‐knockout (AKO) rats ( n = 5–10 per group). Expression (relative to wild‐type rats) of HIF target genes (D) Vegfa (encoding vascular endothelial growth factor a), (E) Arg1 (encoding arginase 1), ( n = 7–8 per group). (F) Protein carbonyl levels in heart homogenates, measured using immunoblotting ( n = 6 per group). One‐way ANOVA with post hoc Tukey's multiple comparison test. Data represented as mean ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: A competitive ELISA assay kit (EIA‐AMY‐1, Ray‐biotech) was used to measure the amylin content in whole heart homogenates from age‐matched male WT, HIP, and UCD rats.

Techniques: Expressing, Immunohistochemistry, Staining, Enzyme-linked Immunosorbent Assay, Knock-Out, Western Blot, Comparison

Journal: bioRxiv

Article Title: Natriuretic Peptide Augmentation Attenuates Renin Cell Hyperactivation and Afferent Arteriolar Hypertrophy During Long-Term Renin-Angiotensin System Inhibition

doi: 10.64898/2026.07.21.739692

Figure Lengend Snippet: A , Experimental scheme for atrial natriuretic peptide (ANP) treatment in As4.1 cells. ANP was added to the culture medium at 0 h and re-administered at 12 h, and the cells and culture supernatants were collected 24 h after the initial treatment. B , Relative Ren1 mRNA expression 24 h after treatment with ANP at the indicated concentrations. Ren1 expression was significantly suppressed by ANP in a dose-dependent manner (n = 6; one-way ANOVA followed by Tukey’s multiple comparison test). C , Renin concentrations in culture supernatants measured via ELISA after treatment with ANP (1 nM) or vehicle control (n = 3; Welch’s t-test). D, Experimental scheme of ANP treatment in the presence or absence of the natriuretic peptide receptor A antagonist A71915. E , Relative Ren1 mRNA expression 24 h after treatment with ANP (1 nM) with or without A71915 (5 µM). A71915 significantly attenuated the inhibitory effect of ANP on Ren1 expression (n = 6; one-way ANOVA, followed by Tukey’s multiple comparison test). Data are shown as mean ± SD. The cell culture data in B, C, and E are representative of three independent experiments.

Article Snippet: Plasma ANP levels were measured using an ANP ELISA kit (EIA-ANP-1; RayBiotech, Norcross, GA, USA).

Techniques: Expressing, Comparison, Enzyme-linked Immunosorbent Assay, Control, Cell Culture

Journal: bioRxiv

Article Title: Natriuretic Peptide Augmentation Attenuates Renin Cell Hyperactivation and Afferent Arteriolar Hypertrophy During Long-Term Renin-Angiotensin System Inhibition

doi: 10.64898/2026.07.21.739692

Figure Lengend Snippet: A, Schematic overview of the long-term renin-angiotensin system (RAS) inhibition experiment in mice. Eight-week-old C57BL/6 mice received once-daily oral gavage of vehicle control (saline), the angiotensin II receptor blocker (ARB) valsartan, or the angiotensin receptor-neprilysin inhibitor (ARNI) sacubitril/valsartan. Blood and kidney samples were collected 24 weeks after treatment initiation. B, Systolic and diastolic blood pressure after 24 weeks of treatment, measured under isoflurane anesthesia using the tail-cuff method. After treatment, blood pressure was significantly lower in both the ARB and ARNI groups than in the control group, with no significant difference between the ARB and ARNI groups (n = 14 for ARB; n = 16 for control and ARNI; one-way ANOVA followed by Tukey’s multiple comparison test). C, Serum blood urea nitrogen (BUN) levels after 24 weeks of treatment. The BUN levels were significantly higher in the ARB group (n = 14 for ARB, n = 16 for control and ARNI; one-way ANOVA followed by Tukey’s multiple comparison test). D, Plasma cystatin C levels measured using ELISA. Plasma cystatin C levels were significantly elevated in the ARB group but not in the ARNI group (n = 10; one-way ANOVA followed by Tukey’s multiple comparison test). E, Relative mRNA expression of renal injury and fibrosis markers in kidney tissues. Long-term ARB treatment significantly increased the expression of Lcn2 , Havcr1 , Col1a1 , and Col3a1 , whereas these increases were attenuated in the ARNI group compared to those in the ARB group (n = 10; one-way ANOVA followed by Tukey’s multiple comparison test). F, Representative Sirius red-stained renal cortical sections and quantification of the fibrotic area. The ARB group showed a significant increase in renal fibrosis, whereas no significant increase was observed in the ARNI group. Scale bar, 100 µm. sBP, systolic blood pressure; dBP, diastolic blood pressure.

Article Snippet: Plasma ANP levels were measured using an ANP ELISA kit (EIA-ANP-1; RayBiotech, Norcross, GA, USA).

Techniques: Inhibition, Control, Saline, Comparison, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Expressing, Staining

Journal: bioRxiv

Article Title: Natriuretic Peptide Augmentation Attenuates Renin Cell Hyperactivation and Afferent Arteriolar Hypertrophy During Long-Term Renin-Angiotensin System Inhibition

doi: 10.64898/2026.07.21.739692

Figure Lengend Snippet: A, Plasma atrial natriuretic peptide (ANP) levels measured using ELISA. Plasma ANP levels significantly increased only in the ARNI group (n = 9 for control and ARB; n = 6 for ARNI; one-way ANOVA followed by Tukey’s multiple comparison test). B, Relative Ren1 mRNA expression in kidney tissues. Long-term ARB treatment markedly increased Ren1 expression, whereas this upregulation was significantly attenuated in the ARNI group compared to that in the ARB group (n = 10; one-way ANOVA followed by Tukey’s multiple comparison test). C, Plasma renin concentrations measured using ELISA. Plasma renin levels were significantly elevated in both the ARB and ARNI groups compared to the control group, although the increase was significantly attenuated in the ARNI group compared to the ARB group. (n = 10; one-way ANOVA followed by Tukey’s multiple comparison test). D, Representative renin immunohistochemistry in mouse kidneys after 24 weeks of treatment. Scale bar, 50 µm. E, quantification of the juxtaglomerular (JG) index. The JG index, defined as the percentage of renin-positive JG apparatuses among the total glomeruli, was significantly increased by long-term RAS inhibition; this increase was attenuated in the ARNI group compared to that in the ARB group (n = 12, one-way ANOVA followed by Tukey’s multiple comparison test). F, Representative α-smooth muscle actin (α-SMA) immunohistochemistry and Masson’s trichrome staining of renal tissues. Scale bar, 50 µm. G, Quantification of afferent arteriolar wall thickness. Wall thickness was calculated as half the difference between the outer and inner diameters of the afferent arteriole. Long-term RAS inhibition significantly increased afferent arteriolar wall thickness, whereas this increase was attenuated in the ARNI group compared to the ARB group (n = 12; one-way ANOVA followed by Tukey’s multiple comparison test). The lower panel shows the relative frequency distribution of afferent arteriolar wall thickness measurements in each treatment group (control, blue; ARB, red; ARNI, green). H, Immunofluorescence staining for α-SMA and renin in afferent arteriolar/JG regions. The left panel shows α-SMA, the middle panel shows renin, and the right panel shows the merged image. Scale bar, 50 µm.

Article Snippet: Plasma ANP levels were measured using an ANP ELISA kit (EIA-ANP-1; RayBiotech, Norcross, GA, USA).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control, Comparison, Expressing, Immunohistochemistry, Inhibition, Staining, Immunofluorescence

Journal: bioRxiv

Article Title: Patolakaturohiniyadi Kashayam , exerts anti-steatotic and anti-obesogenic effects via coordinated regulation of lipid metabolism, inflammation, and incretin signalling

doi: 10.64898/2026.07.14.738366

Figure Lengend Snippet: (A) Representative H&E-stained sections of pancreas (400x); (B) Oral Glucose Tolerance Test (OGTT) showing blood glucose levels at 0, 15-, 30-, 60-, and 120-minutes post-glucose administration; (C) Area Under the Curve (AUC) analysis of OGTT; (D) Plasma insulin levels; (E) Plasma GLP-1 levels. All data are presented as mean□±□SEM (n□=□5); *p□<□0.05, **p□<□0.01, ***p□<□0.001.

Article Snippet: Dulbecco’s modified Eagle’s medium; fetal bovine serum (FBS); and penicillin/streptomycin (PS) (GIBCO, Grand Island, New York); Non-esterified fatty acid free bovine serum albumin (BSA) (Genei, Bangalore, India); triglyceride kit (BeneSphera, Avantor, USA); ascorbic Acid; Folin-Ciocalteu reagent; gallic acid; sodium palmitate (PA); Oil-Red-O (ORO) stain; IBMX; dexamethasone; insulin (Sigma-Aldrich, St. Louis, MO, USA); SPLASH ® LIPIDOMIX ® Mass Spec Standard (Avanti polar lipids, Alabaster, AL, USA, 330707); Pioglitazone hydrochloride tablet (USV Private Limited, Mumbai, India, PIOZ 15); GLP-1 kit (RayBiotech Life Inc., Peachtree Corners, GA, USA, EIA-GLP1); Insulin Elisa kit (Krishgen biosystems, Mumbai, India, KLR0707); AST kit (Elabscience, Texas, USA, E-BC-K236-M); ALT kit (Elabscience, Texas, USA, E-BC-K235-M).

Techniques: Staining, Clinical Proteomics