igf 1  (Novus Biologicals)

Journal: Aging Cell

Article Title: IGF ‐1/ SLC25A33 Maintains Blood‐Labyrinth Barrier Integrity by Suppressing TGF ‐β/Smad2‐Mediated Pericyte Activation in Age‐Related Hearing Loss

doi: 10.1111/acel.70712

Figure Lengend Snippet: Transcriptomic analysis identifies IGF‐1/SLC25A33 dysregulation and TGF‐β pathway activation in aged pericytes, which may be associated with ARHL. (A) Representative click‐evoked ABR waveforms showing a threshold of 30 dB SPL (red trace, left) in a young mouse versus 60 dB SPL (red trace, right) in an aged mouse. (B) Click‐evoked ABR thresholds are significantly elevated in old mice compared to young controls. (C) Frequency‐specific ABR thresholds are markedly increased across all tested frequencies (4–32 kHz) in aged mice. (D) DPOAE thresholds are significantly higher in old mice at F 2 = 17.44 kHz ( n = 6 mice). (E) RNA sequencing of FACS‐purified pericytes from young and aged stria vascularis reveals significant downregulation of Igf1 and Slc25a33 mRNA in aged mice ( n = 6 mice). (F) The most enriched DO terms for the differentially expressed genes when old vs. young. (G) The most enriched GO terms for the downregulated genes when old versus young. (H) The most enriched Reactome terms for the downregulated genes when old versus young. (I) KEGG pathway enrichment analysis highlights the TGF‐β signaling pathway as one of the most differentially activated pathways. (J) Gene–gene interaction network for the downregulated genes when old versus young.

Article Snippet: Sections were blocked in 10% goat serum + 0.3% Triton X‐100 in PBS for 1 h. Primary antibodies were applied overnight at 4°C: IGF‐1 (NOVUS, SPM406, 1:200), SLC25A33 (Bioss, bs‐21231R, 1:200), PDGFRβ (CST, #3169, 1:200), E‐cadherin (CST, #14472, 1:200), α‐SMA (Bioss, bs‐10196R, 1:200), TGFβ1 (Bioworld, BS80030, 1:50), p‐Smad2 (Bioworld, bs‐3419R, 1:500).

Techniques: Analysis, Activation Assay, RNA Sequencing, Purification

Journal: Aging Cell

Article Title: IGF ‐1/ SLC25A33 Maintains Blood‐Labyrinth Barrier Integrity by Suppressing TGF ‐β/Smad2‐Mediated Pericyte Activation in Age‐Related Hearing Loss

doi: 10.1111/acel.70712

Figure Lengend Snippet: Downregulation of IGF‐1 and SLC25A33 are confirmed in the aged stria vascularis. In vitro studies reveal IGF‐1 regulates SLC25A33 expression. (A, B) qPCR shows downregulation of Igf1 and Slc25a33 mRNA expression in aged pericytes ( n = 3 mice); (C) Immunofluorescence staining confirms colocalization of IGF‐1 (green), SLC25A33 (red) and PDGFRβ (purple, pericyte marker) in the stria vascularis. Age‐dependent alteration of IGF‐1 and SLC25A33 were markedly occurred in pericytes, as indicated by arrows. (D‐E) Quantitative analysis reveals significant downregulation of both IGF‐1 and SLC25A33 fluorescence intensity in aged mice relative to young controls ( n = 5, five typical sections from five individual mice were analyzed per group). (F) qPCR shows that inhibition of IGF‐1 signaling (via NVP‐AEW541) significantly reduces Slc25a33 mRNA levels. (G) Representative Western blot of SLC25A33 protein expression. (H) Densitometric quantification demonstrates reduced SLC25A33 protein upon IGF‐1 inhibition. Control: HBVPs without treatment; IGF‐1 (−): HBVPs treated with NVP‐AEW541; IGF‐1 (+): Cells treated with recombinant IGF‐1. Data are mean ± SEM; n = 3 mice.

Article Snippet: Sections were blocked in 10% goat serum + 0.3% Triton X‐100 in PBS for 1 h. Primary antibodies were applied overnight at 4°C: IGF‐1 (NOVUS, SPM406, 1:200), SLC25A33 (Bioss, bs‐21231R, 1:200), PDGFRβ (CST, #3169, 1:200), E‐cadherin (CST, #14472, 1:200), α‐SMA (Bioss, bs‐10196R, 1:200), TGFβ1 (Bioworld, BS80030, 1:50), p‐Smad2 (Bioworld, bs‐3419R, 1:500).

Techniques: In Vitro, Expressing, Immunofluorescence, Staining, Marker, Analysis, Fluorescence, Inhibition, Western Blot, Control, Recombinant

Journal: Aging Cell

Article Title: IGF ‐1/ SLC25A33 Maintains Blood‐Labyrinth Barrier Integrity by Suppressing TGF ‐β/Smad2‐Mediated Pericyte Activation in Age‐Related Hearing Loss

doi: 10.1111/acel.70712

Figure Lengend Snippet: IGF‐1 upregulation promotes SLC25A33 expression in the stria vascularis of aged mice. (A) Immunofluorescence images show IGF‐1 (green) and SLC25A33 (red) expression in the stria vascularis. (B, C) Quantitative analysis reveals that exogenous IGF‐1 supplementation increases both IGF‐1 and SLC25A33 fluorescence intensity, while IGF‐1 inhibition further reduces their expression ( n = 5, five typical sections from five individual mice were analyzed per group). (D) qPCR confirms exogenous IGF‐1 supplementation upregulates Slc25a33 mRNA levels, whereas IGF‐1 inhibition decreases its expression ( n = 3 mice). Control: wild‐type C57BL/6J mice without treatment; IGF‐1 (−): mice treated with NVP‐AEW541; IGF‐1 (+): mice treated with recombinant IGF‐1. Data are mean ± SEM.

Article Snippet: Sections were blocked in 10% goat serum + 0.3% Triton X‐100 in PBS for 1 h. Primary antibodies were applied overnight at 4°C: IGF‐1 (NOVUS, SPM406, 1:200), SLC25A33 (Bioss, bs‐21231R, 1:200), PDGFRβ (CST, #3169, 1:200), E‐cadherin (CST, #14472, 1:200), α‐SMA (Bioss, bs‐10196R, 1:200), TGFβ1 (Bioworld, BS80030, 1:50), p‐Smad2 (Bioworld, bs‐3419R, 1:500).

Techniques: Expressing, Immunofluorescence, Analysis, Fluorescence, Inhibition, Control, Recombinant

Journal: Aging Cell

Article Title: IGF ‐1/ SLC25A33 Maintains Blood‐Labyrinth Barrier Integrity by Suppressing TGF ‐β/Smad2‐Mediated Pericyte Activation in Age‐Related Hearing Loss

doi: 10.1111/acel.70712

Figure Lengend Snippet: Exogenous IGF‐1 rescues blood‐labyrinth barrier integrity and improves hearing function in aged mice. (A) TEM images reveal ultrastructural changes in the stria vascularis. In the IGF‐1‐inhibited group, pericytes exhibit abnormal morphology, loose association with endothelial cells, and widened tight junctions (yellow arrows). These defects are ameliorated by exogenous IGF‐1 administration. (B) Click‐evoked ABR thresholds are significantly elevated in IGF‐1‐suppressed mice but partially rescued by IGF‐1 supplementation. (C–E) At 12 kHz, ABR thresholds are significantly lower in the IGF‐1‐treated group compared to control and suppressed groups, indicating improved auditory sensitivity. (F) Proposed model of IGF‐1/SLC25A33‐mediated regulation of pericyte phenotype and BLB integrity in age‐related hearing loss. Under physiological conditions, pericytes maintain a quiescent state characterized by low α‐SMA and high E‐cadherin expression, contributing to BLB stability. With aging, declining IGF‐1 levels lead to reduced SLC25A33 expression, resulting in mtROS accumulation and activation of the TGF‐β/Smad2 signaling pathway. This drives pericyte transition toward a contractile, pro‐fibrotic phenotype (high α‐SMA, low E‐cadherin), disrupting BLB integrity and promoting cochlear dysfunction. Targeting this axis may offer therapeutic potential for ARHL. Control: wild‐type C57BL/6J mice without treatment; IGF‐1 (−): mice treated with NVP‐AEW541; IGF‐1 (+): mice treated with recombinant IGF‐1. Data are mean ± SEM; n = 6 mice.

Article Snippet: Sections were blocked in 10% goat serum + 0.3% Triton X‐100 in PBS for 1 h. Primary antibodies were applied overnight at 4°C: IGF‐1 (NOVUS, SPM406, 1:200), SLC25A33 (Bioss, bs‐21231R, 1:200), PDGFRβ (CST, #3169, 1:200), E‐cadherin (CST, #14472, 1:200), α‐SMA (Bioss, bs‐10196R, 1:200), TGFβ1 (Bioworld, BS80030, 1:50), p‐Smad2 (Bioworld, bs‐3419R, 1:500).

Techniques: Morphology, Control, Sensitivity, Expressing, Stability, Activation Assay, Recombinant

Journal: Frontiers in Physiology

Article Title: TPM-like manipulations exerted an antidepressant effect and regulated amino acid metabolism and GHR/IGF-1 pathway in the liver in adolescent female CUMS rats

doi: 10.3389/fphys.2026.1752608

Figure Lengend Snippet: TPM-like manipulations increased the expression of IGF-1 and IGF-1R as well as the inflammation in the hippocampus of CUMS-exposed adolescent female rats. (A) Representative bands of IGF-1 and IGF-1R detected by WB. (B) Protein expressions of IGF-1 and IGF-1R in the hippocampus detected by WB. (C) IGF-1 mRNA expression in the hippocampus assessed by qRT-PCR. (D) IGF-1R mRNA expression in the hippocampus assessed by qRT-PCR. (E) TNF-α mRNA expression in the hippocampus assessed by qRT-PCR. (F) IL-1β mRNA expression in the hippocampus assessed by qRT-PCR. All data were analyzed using one-way ANOVA followed by LSD post hoc tests. Data are presented as mean ± SEM (n = 4 for qRT-PCR; n = 5 for WB). * P < 0.05, ** P < 0.01 vs. the CON group. # P < 0.05, ## P < 0.01 vs. the CUMS group. CON, control; CUMS, chronic unpredictable mild stress; TPM, traditional pediatric massage-like manipulations; FLX, fluoxetine; IGF-1, insulin-like growth factor-1; IGF-1R, insulin-like growth factor-1 receptor; TNF-α, tumor necrosis factor alpha; IL-1β, interleukin-1β; WB, Western blotting; qRT-PCR, Quantitative reverse transcription Polymerase Chain Reaction.

Article Snippet: The blots were blocked for 20 minutes at room temperature and incubated overnight at 4 °C with the following primary antibodies: GAPDH (1:10000, A19056, ABclonal), GHR (1:1000, DF8425, Affinity), IGF-1 (1:1000, A24744, ABclonal), and IGF-1R (1:2000, A0243, ABclonal).

Techniques: Expressing, Quantitative RT-PCR, Control, Western Blot, Reverse Transcription, Polymerase Chain Reaction

Journal: Frontiers in Physiology

Article Title: TPM-like manipulations exerted an antidepressant effect and regulated amino acid metabolism and GHR/IGF-1 pathway in the liver in adolescent female CUMS rats

doi: 10.3389/fphys.2026.1752608

Figure Lengend Snippet: TPM-like manipulations maintained GHR/IGF-1 signaling activity in the liver in CUMS-exposed adolescent female rats. (A) Representative bands of GHR and IGF-1 detected by WB. (B) Protein expressions of GHR and IGF-1 in the liver detected by WB. (C) GHR mRNA expression in the liver assessed by qRT-PCR. (D) IGF-1 mRNA expression in the liver assessed by qRT-PCR. (E) Plasma IGF-1 levels in the liver assessed by ELISA. (F) Liver inflammation score. (G) Representative HE-stained liver sections. Representative low-magnification images (100×, scale bar = 200 μm) and corresponding high-magnification insets (200×, scale bar = 100 μm) are shown. “P” indicates the portal area, Arrow “a” indicates inflammatory cell infiltration, and arrow “b” indicates hemorrhage. (H) TNF-α mRNA expression in the liver assessed by qRT-PCR. (I) IL-1β mRNA expression in the liver assessed by qRT-PCR. (J) Plasma ALT levels assessed by ELISA. (K) Plasma AST levels assessed by ELISA. All data were analyzed using one-way ANOVA followed by LSD post hoc tests. Data are presented as mean ± SEM (n = 4 for qRT-PCR; n = 5 for western blotting; n = 7–8 for ELISA and liver inflammation scoring). * P < 0.05, ** P < 0.01 vs. the CON group. # P < 0.05, ## P < 0.01 vs. the CUMS group. CON, control; CUMS, chronic unpredictable mild stress; TPM, traditional pediatric massage-like manipulations; FLX, fluoxetine; GHR, growth hormone receptor; IGF-1, insulin-like growth factor-1; TNF-α, tumour necrosis factor alpha; IL-1β, interleukin-1β; ALT, alanine aminotransferase; AST, aspartate aminotransferase; qRT-PCR, Quantitative reverse transcription Polymerase Chain Reaction; ELISA, Enzyme-linked immunosorbent assay.

Article Snippet: The blots were blocked for 20 minutes at room temperature and incubated overnight at 4 °C with the following primary antibodies: GAPDH (1:10000, A19056, ABclonal), GHR (1:1000, DF8425, Affinity), IGF-1 (1:1000, A24744, ABclonal), and IGF-1R (1:2000, A0243, ABclonal).

Techniques: Activity Assay, Expressing, Quantitative RT-PCR, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Control, Reverse Transcription, Polymerase Chain Reaction

Journal: Frontiers in Physiology

Article Title: TPM-like manipulations exerted an antidepressant effect and regulated amino acid metabolism and GHR/IGF-1 pathway in the liver in adolescent female CUMS rats

doi: 10.3389/fphys.2026.1752608

Figure Lengend Snippet: Spearman correlation matrix of hepatic metabolic changes, behavioral outcomes, and IGF-1-related protein expression in the liver and hippocampus. The heatmap illustrates the Spearman’s rank correlation coefficients (r) between the relative abundance of the 22 key differential hepatic metabolites and seven key outcome variables: behavioral parameters including sucrose preference percentage, the time spent in the center zone in the open field test, and the number of platform crossings in the Morris water maze probe trial, hepatic protein expression of growth hormone receptor and insulin-like growth factor 1, and the protein expression of insulin-like growth factor 1 and insulin-like growth factor 1 receptor in the hippocampus. The color scale indicates the strength and direction of the correlation, with red representing a positive correlation and blue representing a negative correlation. Asterisks denote the level of statistical significance. (n = 5). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The blots were blocked for 20 minutes at room temperature and incubated overnight at 4 °C with the following primary antibodies: GAPDH (1:10000, A19056, ABclonal), GHR (1:1000, DF8425, Affinity), IGF-1 (1:1000, A24744, ABclonal), and IGF-1R (1:2000, A0243, ABclonal).

Techniques: Expressing

Journal: Frontiers in Physiology

Article Title: TPM-like manipulations exerted an antidepressant effect and regulated amino acid metabolism and GHR/IGF-1 pathway in the liver in adolescent female CUMS rats

doi: 10.3389/fphys.2026.1752608

Figure Lengend Snippet: Correlation analysis of hepatic L-aspartic acid changes with circulating IGF-1 concentration, liver injury-related indices, and depression-related behaviors. (A) L-Aspartic Acid concentration in the liver assessed by ELISA (B) Correlation analysis between liver L-Aspartic Acid and plasma IGF-1. (C) Correlation analysis between liver L-Aspartic Acid and sucrose preference percentage in the sucrose preference test. (D) Correlation analysis between liver L-Aspartic Acid and plasma AST. (E) Correlation analysis between liver L-Aspartic Acid and plasma ALT. Hepatic L-Aspartic Acid data were analyzed using one-way ANOVA followed by LSD post hoc tests. Pearson correlation analysis was performed for panels B–E. Data are presented as mean ± SEM ( n = 7–8 for ELISA; n = 28 for Pearson correlation analysis). * P < 0.05, ** P < 0.01 vs. the CON group. # P < 0.05, ## P < 0.01 vs. the CUMS group. CON, control; CUMS, chronic unpredictable mild stress; TPM, traditional pediatric massage-like manipulations; FLX, fluoxetine; IGF-1, insulin-like growth factor-1; AST, aspartate aminotransferase; ALT, alanine aminotransferase; ELISA, Enzyme-linked immunosorbent assay.

Article Snippet: The blots were blocked for 20 minutes at room temperature and incubated overnight at 4 °C with the following primary antibodies: GAPDH (1:10000, A19056, ABclonal), GHR (1:1000, DF8425, Affinity), IGF-1 (1:1000, A24744, ABclonal), and IGF-1R (1:2000, A0243, ABclonal).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Control