Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Induction of IL‐20 expression in NP cells by resistin stimulation. RNA samples were isolated at the indicated time points or doses and subjected to real‐time polymerase chain reaction analysis. Data are presented as fold changes in fluorescent density from control NP cells (CL) normalized to 18S rRNA level (A, B). The IL‐20 protein in conditioned media was detected by enzyme‐linked immunosorbent assay (C, D). NP cells were kept as controls (CL) or stimulated with 50 ng/mL resistin for the times indicated (C), or the cells were stimulated with resistin at various doses for 4 h (D). Data are shown as mean ± standard error of the mean (SEM). * p < 0.05 versus control NP cells (CL).

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Expressing, Isolation, Real-time Polymerase Chain Reaction, Control, Enzyme-linked Immunosorbent Assay

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: p38 MAPK and Akt pathways are required for resistin‐induced expression of IL‐20. (A, B) NP cells were kept as control (CL) or stimulated with 50 ng/mL resistin for 2 h (A) and 4 h (B). Before being kept as CL or stimulated with resistin, NP cells were pretreated with PD98059 (PD), SP600125 (SP), SB203580 (SB), or LY294002 (LY) individually for 1 h. (A) RNA was isolated and subjected to real‐time polymerase chain reaction analysis. Data are presented as fold changes in fluorescent density from control NP cells (CL) normalized to 18S rRNA level. (B) The IL‐20 protein secretion in conditioned media was determined by enzyme‐linked immunosorbent assay. The results are shown as mean ± standard error of the mean. * p < 0.05 versus CL. # p < 0.05 versus vehicle control (dimethyl sulfoxide) with resistin stimulation. (C) NP cells were employed as CL or stimulated with resistin for the durations indicated, and p38 and Akt phosphorylations were determined by Western blotting.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Expressing, Control, Isolation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Induction of NF‐kB‐p65 activity by resistin stimulation in NP cells. (A) IL‐20 mRNA expression levels were determined in NP cells pretreated with vehicle (DMSO) or SN50, or transfected with control siRNA (si‐CL) or si‐p65, and then stimulated with 50 ng/mL resistin for 2 h. RNA samples were isolated and subjected to real‐time polymerase chain reaction analysis. Data are presented as fold changes in fluorescent density from control NP cells (CL) normalized to 18S rRNA level. (B, C) The NF‐kB p65 activation was determined by a transcription factor (TF)‐enzyme‐linked immunosorbent assay. (B) NP cells were employed as control (CL) or stimulated with resistin for the durations indicated. (C) NP cells were pretreated individually with PD98059 (PD), SP600125 (SP), SB203580 (SB), or LY294002 (LY) for 1 h before being used as controls (CL) or stimulated with 50 ng/mL resistin for 1 h. NF‐κB p65 activity was then analyzed. All bar graphs represent folds of CL NP cells, mean ± standard error of the mean. * p < 0.05 versus CL. # p < 0.05 versus DMSO or si‐CL under resistin stimulation.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Activity Assay, Expressing, Transfection, Control, Isolation, Real-time Polymerase Chain Reaction, Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Blockade of TLR4 activity inhibited resistin‐induced IL‐20 expression. NP cells were kept as controls (CL), or pretreated with isotype‐matched IgG (Ab‐IgG) and specific TLR4 neutralizing antibody (Ab‐TLR4), or transfected with the control siRNA (si‐CL) and si‐TLR4, and subsequently stimulated with resistin for 2 h (A) and 1 h (B). (A) IL‐20 mRNA levels were determined through real‐time PCR in NP cells and normalized to 18S rRNA. (B) The activation of NF‐kB‐p65 in NP cells after resistin stimulation was analyzed using transcription factor (TF) ELISA. All bar graphs represent folds of control NP cells (CL), mean ± standard error of the mean. * p < 0.05 versus CL NP cells. # p < 0.05 versus IgG‐pretreated or si‐CL‐transfected NP cells under resistin stimulation.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Activity Assay, Expressing, Transfection, Control, Real-time Polymerase Chain Reaction, Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Pre‐exposure of NP cells to 5% cyclic stretch with 0.1 Hz for 30 min inhibited resistin‐induced IL‐20 expression. Static NP cells were stimulated with resistin without prestretching (static). NP cells were kept as controls (CL) or pre‐exposed to cyclic stretch at 5% with 0.1 Hz for the indicated durations followed by resistin stimulation. (A) The mRNA levels of IL‐20 in NP cells were determined through real‐time polymerase chain reaction and normalized to 18S rRNA. * p < 0.05 versus CL NP cells. ** p < 0.05 versus static NP cells with resistin stimulation. # p < 0.05 versus resistin‐treated NP cells with cyclic stretch at 5% with 0.1 Hz for 10' and 1 h. (B) The phosphorylation of p38 MAPK and Akt was determined by Western blotting. (C) NF‐kB‐p65 activation in NP cells after 1 h resistin stimulation was analyzed by TF‐ELISA. All bar graphs represent folds of control NP cells (CL), mean ± standard error of the mean. * p < 0.05 versus CL NP cells. # p < 0.05 versus static NP cells with resistin stimulation.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Phospho-proteomics, Western Blot, Activation Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Upregulation of NRF2 inhibited resistin‐induced IL‐20 expression in NP cells. (A) NP cells were used as static control (static) or exposed to 5% with 0.1 Hz cyclic stretch for 30 min or 2 h. The expression of NRF2 in the nucleus was determined by Western blotting. (B–D) NP cells were used as static control (static), or pre‐exposed to 5% with 0.1 Hz for 30 min, and then treated with resistin (50 ng/mL) for 2 h (B), 30 min (C), and 1 h (D). Prior to cyclic stretch exposure, NP cells were transfected with the control siRNA (si‐CL) or si‐NRF2. (B) The levels of IL‐20 mRNA in NP cells were determined through real‐time polymerase chain reaction and normalized to 18S rRNA. (C) The phosphorylation of p38 MAPK, and Akt was determined by Western blotting. (D) NF‐kB‐p65 activation in NP cells was analyzed by TF‐ELISA. All bar graphs represent the percentage of static NP cells (static), mean ± standard error of the mean. * p < 0.05 versus static NP cells.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Expressing, Control, Western Blot, Transfection, Real-time Polymerase Chain Reaction, Phospho-proteomics, Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Schematic representation of the signaling pathways regulating 5% with 0.1 Hz cyclic stretch‐induced NRF2 expression and consequent inhibition of resistin effect in human NP cells.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Protein-Protein interactions, Expressing, Inhibition

Journal: Journal of Translational Medicine

Article Title: Multi-omics analysis reveals a feedback loop amplifying immune responses in acute graft-versus-host disease due to imbalanced gut microbiota and bile acid metabolism

doi: 10.1186/s12967-024-05577-x

Figure Lengend Snippet: Granulocyte-like cells derived from patients with non-aGVHD exhibited elevated levels of RETN expression, suggesting a potential role for resistin in preventing aGVHD. A Heatmap display of the communication probability of each cluster within the combined dataset of PBMCs from patients undergoing allo-HSCT. B RETN and its corresponding receptor CAP1 were expresed in granulocyte-like cells across each sample. C Representative immunofluorescence staining of CD45 and resistin in the small intestine of control or aGVHD mouse model groups. Scale bar, 100 µm. D IL-1β concentrations were measured using ELISA assay in cell culture supernatants (n = 3) following various treatments involving LPS and resistin. E Venn diagrams illustrating the shared conserved downregulated genes among distinct lineages (group 1: neutrophils vs. macrophages vs. monocytes; group 2: cDC1 vs. cDC2 vs. migratory DC; group 3: plasmacytoid dendritic cell (pDC) vs. B cells; group 4: CD4 + T cells vs. CD8 + T cells vs. Treg; group 5: innate lymphoid cell (ILC) vs. γδ T cells vs. NK cells). F Summary of the resistin-driven immune regulation in the presence of LPS or other special conditions

Article Snippet: Following PMA stimulation, the culture medium was replaced with RPMI1640 supplemented with 10% FBS, 1 × P/S, and LPS (1, 10, or 100 ng/mL) (Cat# L8880, Salarbio) or recombinant human resistin (1, 10, and 100 ng/mL) (Cat# CJ48, novoprotein) for 24 h. The resulting cell-free supernatants were harvested and centrifuged at 500× g for 10 min.

Techniques: Derivative Assay, Expressing, Immunofluorescence, Staining, Control, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Journal of Translational Medicine

Article Title: Multi-omics analysis reveals a feedback loop amplifying immune responses in acute graft-versus-host disease due to imbalanced gut microbiota and bile acid metabolism

doi: 10.1186/s12967-024-05577-x

Figure Lengend Snippet: A simplified schematic diagram illustrating the dysregulated gut microbiota and bile acid metabolism positive feedback loop, which exacerbates immune responses and exacerbates aGVHD. Improper treatment of patients undergoing allo-HSCT alters the gut microbiota and disrupts the imbalanced bile acid metabolism. Dysregulated bile acid metabolism results in reduced production of IL1RN, defensins, and resistin. Deficiency of defensins and resistin fails to inhibit harmful microorganisms, leading to increased production of proinflammatory cytokines and proteins, such as IL-17A, IL-1β, TNFα, IL-6, IL-12, and members of the S100A family. Additionally, it was determined that resistin can inhibit aGVHD by downregulating the expression of IL1B, DUSP1, and members of the AP-1 family

Article Snippet: Following PMA stimulation, the culture medium was replaced with RPMI1640 supplemented with 10% FBS, 1 × P/S, and LPS (1, 10, or 100 ng/mL) (Cat# L8880, Salarbio) or recombinant human resistin (1, 10, and 100 ng/mL) (Cat# CJ48, novoprotein) for 24 h. The resulting cell-free supernatants were harvested and centrifuged at 500× g for 10 min.

Techniques: Expressing

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Effects of resistin on porcine ovarian follicle steroidogenesis in prepubertal animals: an in vitro study

doi: 10.1186/1477-7827-11-45

Figure Lengend Snippet: Effects of recombinant human resistin (0.1, 1, 10, and 100 ng/ml) on steroid hormone (i.e., progesterone [P4], androstendione [A4], testosterone [T], and estradiol [E2]) secretion in culture medium from A) small, B) medium and C) large ovarian follicles. ELISA experiments were performed three independent times. In each experiment six ovaries from three different animals were selected. Data were plotted as mean±S.E.M. Different letters indicate statistically significant differences among groups ( P < 0.05).

Article Snippet: Recombinant human resistin was obtained from Phoenix Pharmaceuticals, Inc. (Burlingame, CA, USA).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Effects of resistin on porcine ovarian follicle steroidogenesis in prepubertal animals: an in vitro study

doi: 10.1186/1477-7827-11-45

Figure Lengend Snippet: Effects of recombinant human resistin (0.1, 1, 10, and 100 ng/ml) on the mRNA expression of steroidogenic enzymes (i.e., CYP11A1, 3βHSD, CYP17A1, 17βHSD, and CYP19A1 ) in A) small, B) medium and C) large ovarian follicles. The expression of mRNA has been determined by real time-PCR. The expression of each gene was normalized to the expression of GAPDH. Real time PCR experiments were performed three independent times. In each experiment six ovaries from three different animals were selected. Data were plotted as mean±S.E.M. Statistical significance is indicated by * P < 0.05.

Article Snippet: Recombinant human resistin was obtained from Phoenix Pharmaceuticals, Inc. (Burlingame, CA, USA).

Techniques: Recombinant, Expressing, Real-time Polymerase Chain Reaction

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Effects of resistin on porcine ovarian follicle steroidogenesis in prepubertal animals: an in vitro study

doi: 10.1186/1477-7827-11-45

Figure Lengend Snippet: Effects of recombinant human resistin. (0.1, 1, 10, and 100 ng/ml) on the protein expression of steroidogenic enzymes CYP11A1, 3βHSD, CYP17A1, 17βHSD, and CYP19 small, medium and large ovarian follicles. The representative samples of western blots are shown in the panels. Western blot experiments were repeated three independent times using samples from three different animals. Data were plotted as mean±S.E.M. Statistical significance is indicated by * P < 0.05.

Article Snippet: Recombinant human resistin was obtained from Phoenix Pharmaceuticals, Inc. (Burlingame, CA, USA).

Techniques: Recombinant, Expressing, Western Blot