Journal: Cell Death & Disease

Article Title: C1q-TNF-related protein-3 attenuates pressure overload-induced cardiac hypertrophy by suppressing the p38/CREB pathway and p38-induced ER stress

doi: 10.1038/s41419-019-1749-0

Figure Lengend Snippet: a Representative western blot of CTRP3 levels in different tissues of WT mice and Ctrp3 -KO mice ( n = 3 mice per group). b The HW/BW ratio in animals after 4 weeks of TAC ( n = 5–8 mice per group). c–e The left ventricular ejection fraction (LVEF), IVSd, and LVPWd, accordingly, determined by analyzing the echocardiographic images ( n = 12–14 mice per group). f Representative images of the gross murine heart and sections stained with hematoxylin and eosin (HE), and wheat germ agglutinin (WGA) ( n = 5–8 mice per group). g The mean cross-sectional area of cardiomyocytes from the indicated groups ( n ≥ 100 cells per group). h Representative images of the murine heart sections (after 4 weeks of TAC) stained with Masson stain, arranged with the perivascular area at the top and the interstitial area at the bottom ( n = 5–8 mice per group). i The LV collagen volume in different groups ( n ≥ 40 fields per group). j Real-time polymerase chain reaction (real-time PCR) analysis of the expression of genes encoding hypertrophic markers α-MHC, β-MHC, ANP, BNP, Acta-1, IL-6, and Rcan1.4, and the fibrotic markers TGF-β1, collagen-I, and collagen-III in each group ( n = 6 mice per group). b – j The data were analyzed by one-way ANOVA. ** p < 0.01 vs. SHAM, # p < 0.05 vs. TAC. In the bar graphs, the data are presented as the mean ± standard error of the mean (SEM)

Article Snippet: Some NRCMs were also treated with CTRP3 (5 μg/ml) in the presence of PE for 24 h. Before PE treatment, some NRCMs were pretreated respectively with 1 μM SB203580 or 1 μM 666–15 (CREB inhibitor, purchased from Med Chem Express) for 2 h to inhibit phosphorylation of p38 or CREB .

Techniques: Western Blot, Staining, Real-time Polymerase Chain Reaction, Expressing

Journal: Cell Death & Disease

Article Title: C1q-TNF-related protein-3 attenuates pressure overload-induced cardiac hypertrophy by suppressing the p38/CREB pathway and p38-induced ER stress

doi: 10.1038/s41419-019-1749-0

Figure Lengend Snippet: a Representative western blot and quantification of CTRP3 levels in the heart tissue of WT mice and LV-CTRP3 mice ( n = 3 mice per group). b CTRP3 serum levels in mice infected with LV-NULL or LV-CTRP3 ( n = 3 mice per group). c The HW/BW ratio in animals after 4 weeks of TAC surgery ( n = 5–7 mice per group). d–f LVEF, IVSd, and LVPWd, accordingly, determined by analyzing the echocardiographic images ( n = 12–14 mice per group). g Representative images of the gross murine heart and sections stained with HE and WGA ( n = 5–7 mice per group). h The mean cross-sectional area of cardiomyocytes from the indicated groups ( n ≥ 100 cells per group). i Representative images of the murine heart sections (after 4 weeks of TAC) stained with Masson stain, arranged with the perivascular area at the top and the interstitial area at the bottom ( n = 5–7 mice per group). j The LV collagen volume in different groups ( n ≥ 40 fields per group). k Real-time PCR analysis for the expression of genes encoding the hypertrophic markers α-MHC, β-MHC, ANP, BNP, Acta-1, IL-6, and Rcan1.4, and the fibrotic markers TGF-β1, collagen-I, and collagen-III in each group (n = 6 mice per group). c – k The data were analyzed by one-way ANOVA. * p < 0.05, ** p < 0.01 vs. SHAM, # p < 0.05 vs. TAC; ns, not significant. In the bar graphs, the data are presented as the mean ± SEM

Article Snippet: Some NRCMs were also treated with CTRP3 (5 μg/ml) in the presence of PE for 24 h. Before PE treatment, some NRCMs were pretreated respectively with 1 μM SB203580 or 1 μM 666–15 (CREB inhibitor, purchased from Med Chem Express) for 2 h to inhibit phosphorylation of p38 or CREB .

Techniques: Western Blot, Infection, Staining, Real-time Polymerase Chain Reaction, Expressing

Journal: Cell Death & Disease

Article Title: C1q-TNF-related protein-3 attenuates pressure overload-induced cardiac hypertrophy by suppressing the p38/CREB pathway and p38-induced ER stress

doi: 10.1038/s41419-019-1749-0

Figure Lengend Snippet: a, b Representative western blot (top) and quantification (bottom) of the p38-CREB signaling pathway activity in the hearts of mice with different genotypes (WT, Ctrp3 -KO, and LV-CTRP3) 4 weeks after sham treatment or TAC surgery ( n = 5–6 mice per group). c, d Representative immunofluorescence images of murine heart sections stained with p-p38 (red) and DAPI (blue) (left), and the percentage of p-p38–positive nuclei (right) in the hearts of mice with different genotypes (WT, Ctrp3 -KO, and LV-CTRP3) 4 weeks after sham treatment or TAC surgery ( n = 5–6 mice per group). The data were analyzed by one-way ANOVA. * p < 0.05, ** p < 0.01 vs. SHAM, # p < 0.05 vs. TAC. In the bar graphs, the data are presented as the mean ± SEM

Article Snippet: Some NRCMs were also treated with CTRP3 (5 μg/ml) in the presence of PE for 24 h. Before PE treatment, some NRCMs were pretreated respectively with 1 μM SB203580 or 1 μM 666–15 (CREB inhibitor, purchased from Med Chem Express) for 2 h to inhibit phosphorylation of p38 or CREB .

Techniques: Western Blot, Activity Assay, Immunofluorescence, Staining

Journal: Cell Death & Disease

Article Title: C1q-TNF-related protein-3 attenuates pressure overload-induced cardiac hypertrophy by suppressing the p38/CREB pathway and p38-induced ER stress

doi: 10.1038/s41419-019-1749-0

Figure Lengend Snippet: a Representative western blot (left) and quantification (right) of the p38-CREB signaling pathway activity in NRCMs from the indicated groups ( n = 5 samples per group). b Representative western blot (left) and quantification (right) of the p38-CREB signaling pathway activity in NRCMs from the indicated groups ( n = 5 samples per group). c Left: Representative immunofluorescence images of NRCMs transfected with siRNA or si-CTRP3, and stained with α-actinin (red) and DAPI (blue). The NRCMs were treated with dimethyl sulfoxide (DMSO) or a p38 inhibitor SB203580 (1 μM). Right: The mean cell surface area of NRCMs in the indicated groups ( n ≥ 30 cells per group). d Real-time PCR analysis of the expression of genes encoding the hypertrophic markers β-MHC, ANP, and BNP in the indicated groups ( n = 5 samples per group). e Representative western blot (left), and quantification (right) of p-CREB and CREB levels in NRCMs from the indicated groups ( n = 5 samples per group). f Left: Representative immunofluorescence images of PBS- and CTRP3-treated NRCMs stained with α-actinin (red) and DAPI (blue). The NRCMs were treated with DMSO or SB203580 (1 μM). Right: The mean cell surface area of NRCMs in the indicated groups ( n ≥ 30 cells per group). g Real-time PCR analysis of the expression of genes encoding the hypertrophic markers β-MHC, ANP, and BNP in the indicated groups ( n = 4 samples per group). h Representative western blot (left), and quantification (right) of p-CREB and CREB levels in NRCMs from the indicated groups ( n = 5 samples per group). The data were analyzed by one-way ANOVA. a , and b * p < 0.05, ** p < 0.01 vs. control, # p < 0.05 vs. PE. e , h * p < 0.05 between the two indicated groups; ns, not significant. In the bar graphs, the data are presented as the mean ± SEM

Article Snippet: Some NRCMs were also treated with CTRP3 (5 μg/ml) in the presence of PE for 24 h. Before PE treatment, some NRCMs were pretreated respectively with 1 μM SB203580 or 1 μM 666–15 (CREB inhibitor, purchased from Med Chem Express) for 2 h to inhibit phosphorylation of p38 or CREB .

Techniques: Western Blot, Activity Assay, Immunofluorescence, Transfection, Staining, Real-time Polymerase Chain Reaction, Expressing, Control

Journal: Cell Death & Disease

Article Title: C1q-TNF-related protein-3 attenuates pressure overload-induced cardiac hypertrophy by suppressing the p38/CREB pathway and p38-induced ER stress

doi: 10.1038/s41419-019-1749-0

Figure Lengend Snippet: a Representative western blot of the activity of GRP78, eIF2α, IRE1α, CHOP, and ATF6 in mice hearts from indicated groups. b Statistical diagrams of GRP78, eIF2α, IRE1α, CHOP, and ATF6 protein expression in mice hearts from indicated groups ( n = 6 mice per group). c Representative western blot of the activity of GRP78, eIF2α, IRE1α, CHOP, and ATF6 in mice hearts from indicated groups. d Statistical diagrams of GRP78, eIF2α, IRE1α, CHOP, and ATF6 protein expression in mice hearts from indicated groups ( n = 6 mice per group). e, f Representative immunofluorescence images of murine heart sections stained with GRP78 (red) and DAPI (blue) in the hearts of mice with different genotypes (WT, Ctrp3 -KO, and LV-CTRP3) 4 weeks after sham treatment or TAC surgery ( n = 5–6 mice per group). g Representative western blot (left), and quantification (right) of GRP78, eIF2α, IRE1α, CHOP, and ATF6 in the hearts of mice from the indicated groups ( n = 5 mice per group). The data were analyzed by one-way ANOVA. b , d * p < 0.05, ** p < 0.01 vs. SHAM, # p < 0.05 vs. TAC. g * p < 0.05 and # p < 0.05 between indicated groups. In the bar graphs, the data are presented as the mean ± SEM

Article Snippet: Some NRCMs were also treated with CTRP3 (5 μg/ml) in the presence of PE for 24 h. Before PE treatment, some NRCMs were pretreated respectively with 1 μM SB203580 or 1 μM 666–15 (CREB inhibitor, purchased from Med Chem Express) for 2 h to inhibit phosphorylation of p38 or CREB .

Techniques: Western Blot, Activity Assay, Expressing, Immunofluorescence, Staining

Journal: Cell Death & Disease

Article Title: C1q-TNF-related protein-3 attenuates pressure overload-induced cardiac hypertrophy by suppressing the p38/CREB pathway and p38-induced ER stress

doi: 10.1038/s41419-019-1749-0

Figure Lengend Snippet: a Representative western blot (left) and quantification (right) of the ER stress signaling pathway activity in NRCMs from the indicated groups ( n = 4 samples per group). b Representative western blot (left) and quantification (right) of the p38-CREB and ER stress signaling pathway activity in NRCMs from the indicated groups ( n = 4 samples per group). c Representative western blot (left) and quantification (right) of ER stress and p38-CREB signaling pathway activity, protein expression levels of ANP and β-MHC in NRCMs from the indicated groups ( n = 4 samples per group). * p < 0.05, ** p < 0.01 between indicated groups. In the bar graphs, the data are presented as the mean ± SEM. d Proposed protective mechanism of the role of CTRP3 in pathological cardiac hypertrophy. CTRP3 inhibits the activation of the p38/CREB signaling pathway and ER stress pathway in cardiomyocytes under pressure overload, thereby alleviating pathological cardiac hypertrophy

Article Snippet: Some NRCMs were also treated with CTRP3 (5 μg/ml) in the presence of PE for 24 h. Before PE treatment, some NRCMs were pretreated respectively with 1 μM SB203580 or 1 μM 666–15 (CREB inhibitor, purchased from Med Chem Express) for 2 h to inhibit phosphorylation of p38 or CREB .

Techniques: Western Blot, Activity Assay, Expressing, Activation Assay

Journal: BMJ Open Diabetes Research & Care

Article Title: Plasma C1q/tumor necrosis factor-related protein-3 concentrations are associated with diabetic peripheral neuropathy

doi: 10.1136/bmjdrc-2021-002746

Figure Lengend Snippet: Plasma C1q/tumor necrosis factor-related protein-3 (CTRP3) concentrations in patients without diabetic peripheral neuropathy (non-DPN) and with DPN. Data are presented as means±SD; ***p<0.001 compared with non-DPN.

Article Snippet: The plasma CTRP3 concentrations were determined by ELISA with specific antibody against CTRP3 (Proteintech, Wuhan, China) according to standard protocols.

Techniques: Clinical Proteomics

Journal: BMJ Open Diabetes Research & Care

Article Title: Plasma C1q/tumor necrosis factor-related protein-3 concentrations are associated with diabetic peripheral neuropathy

doi: 10.1136/bmjdrc-2021-002746

Figure Lengend Snippet: The results of the nerve conduction testing from groups divided according to plasma CTRP3 tertiles

Article Snippet: The plasma CTRP3 concentrations were determined by ELISA with specific antibody against CTRP3 (Proteintech, Wuhan, China) according to standard protocols.

Techniques: Clinical Proteomics

Journal: BMJ Open Diabetes Research & Care

Article Title: Plasma C1q/tumor necrosis factor-related protein-3 concentrations are associated with diabetic peripheral neuropathy

doi: 10.1136/bmjdrc-2021-002746

Figure Lengend Snippet: Scatter plots showing the correlations of plasma C1q/tumor necrosis factor-related protein-3 (CTRP3) concentrations with nerve conduction parameters. The correlations were assessed by Pearson’s correlation test. M represents motor, S represents sensory, L represents left, R represents right, T represents tibia, P represents peroneal and Su represents sural.

Article Snippet: The plasma CTRP3 concentrations were determined by ELISA with specific antibody against CTRP3 (Proteintech, Wuhan, China) according to standard protocols.

Techniques: Clinical Proteomics

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: (A) Impacts of increasing CTRP3 concentration on the viability of THP-1 differentiated macrophages and mouse peritoneal macrophages. Cells were suspended into 1 × 10 4 cells /100μL/well; THP-1 cells were induced to differentiate into macrophages by culture with 100 ng/mL PMA for 48 h. Mouse peritoneal macrophages were extracted and cultured as mentioned above, after which increasing doses of CTRP3 (0-10 μg/mL) were added; 24 h later, the CCK-8 assay was used to detect viability (mean ± SD). (B-C) Impacts of CTRP3 on LPS-triggered macrophage lipid deposition and foam cell formation in both kinds of macrophages (1 × 10 6 cells/mL/well). The cholesterol content was detected using a cholesterol quantification kit to evaluate lipid deposition in LPS-triggered macrophages (B, mean ± SD, * : compared to the control group, P < 0.05; # : compared to the ox-LDL and LPS treatment group, P < 0.05). Oil red O staining was performed to analyze the effect of CTRP3 on foam cell formation (C, THP-1 cells: 200×, mouse peritoneal macrophages: 400×). At least three independent experiments were performed.

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Concentration Assay, Cell Culture, CCK-8 Assay, Control, Staining

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: THP-1 cells and mouse peritoneal macrophages (1 × 10 6 cells/mL/well) were preincubated with different concentrations of CTRP3 (0-1.0 μg/ mL) for 30 min. After LPS and ox-LDL stimulation, the supernatant protein levels of TNFα, IL-6, MCP-1, IL-1β and MMP-9 were evaluated by ELISA. At least three independent experiments were performed (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS and ox-LDL treatment group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: Cells (1 × 10 6 cells /mL/well) were treated, and expression of specific M1 and M2 markers on the cell surface was detected by FCM to observe macrophages polarization. For THP-1 cells, cells expressing CD68 but not CD206/MR (CD68 + CD206 − ) were considered to be M1 macrophages; cells co-expressing CD68 and CD206 (CD68 + CD206 + ) were considered to be M2 macrophages. For mouse peritoneal macrophages, cells expressing CCR7 and CD86 were considered to be M1 macrophages; cells expressing CD163 and CD206/MR were considered to be M2 macrophages. The macrophage phenotype ratio was calculated as (CCR7 plus CD86 expression)/(CD163 plus CD206 expression). At least three independent experiments were performed (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS but without CTRP3 treatment group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Expressing, Control

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: PMA-induced THP-1 macrophages (1 × 10 6 cells/mL/well) were divided into three groups: the control group was treated with PBS for 24 h; the LPS group was treated with 100 ng/mL LPS for 24 h; the LPS+CTRP3 group was treated with 1 μg/mL CTRP3 for 30 min before LPS stimulation for 24 h. mRNA expression of specific M1 marker and M2 markers was assessed by qRT-PCR. Relative mRNA concentrations were calculated using the ΔΔCT method, and the expression level was normalized to that of endogenous β-actin. At least three independent experiments were performed (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Control, Expressing, Marker, Quantitative RT-PCR

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: Different concentrations of CTRP3 were added before cells (1 × 10 6 cells/mL/well) were exposed to LPS and ox-LDL, and protein expression of TLR4, MyD88, p-NF-κB p65 and NF-κB p65 was assessed by WB (A-D) . Protein expression of PPARγ, LXRα, ABCA1 and ABCG1 was assessed by WB (E-I) (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS and ox-LDL treatment group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Expressing, Control

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: PMA-induced THP-1 macrophages (1 × 10 6 cells/mL/well) were divided into five groups:the control group was treated with PBS; the LPS group was treated with 100 ng/mL LPS for 24 h prior to 100 μg/mL ox-LDL for 24 h; the LPS+CTRP3 (CT3) group was pretreated with 1 μg/mL CTRP3 for 30 min before LPS and ox-LDL stimulation; the LPS+PDTC group was pretreated with 100 μM PDTC for 1 h and then stimulated with LPS and ox-LDL; the LPS+PDTC+CT3 group was treated with 100 μM PDTC for 1 h and then 1 μg/mL CTRP3 for 30 min before LPS and ox-LDL stimulation. Expression of TLR4, MyD88, NF-κB p-p65, t-p65 and β-actin was detected by WB (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS group, P < 0.05, $ : compared to the LPS+CT3 group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Control, Expressing

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: PMA-induced THP-1 macrophages (1 × 10 6 cells/mL/well) were divided into six groups: the first three group were treated as mentioned above, and the other three groups were transfected with si-ctrl, si-PPARγ or si-LXRα for 48 h, and then treated with 1 μg/mL CTRP3 for 30 min, and finally stimulated with LPS and ox-LDL. Expression of PPARγ, LXRα, ABCA1, ABCG1 and β-actin was detected by WB (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS group, P < 0.05, $ : compared to the LPS+CT3 group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Transfection, Expressing, Control