mouse il 1β enzyme linked immunosorbent assay elisa kit  (Proteintech)

Journal: Journal of Pharmaceutical Analysis

Article Title: Novel bioactive peptides targeting Keap1-Nrf2 interaction for combating UVA-induced skin aging: Computational discovery and experimental validation

doi: 10.1016/j.jpha.2025.101446

Figure Lengend Snippet: Protective effects of Seq1 and Seq3 on ultraviolet A (UVA)-induced cellular aging in human keratinocytes (HaCaT) cells. (A) The effect of peptide treatment on the migration of HaCaT cells. (B) The extent of wound closure was quantified and depicted on a histogram. (C) Assessment of the effects of peptides on senescence associated β-galactosidase (SA-β-Gal) activity in HaCaT cells. (D) Quantitation of SA-β-gal positive cells in HaCaT cells. (E) Immunofluorescence (IF) analysis of phosphorylated γ-H2AX in response to UVA irradiation and peptide treatment at varying concentrations to determine their influence. (F) 2’,7’-Dichlorodihydrofluorescein diacetate (DCFH-DA) staining reveals intracellular reactive oxygen species (ROS) levels in UVA-irradiated HaCaT cells following peptide or tert-butylhydroquinone (t-BHQ) treatment. (G) Fluorescence intensity of ROS. (H) Western blot analysis of Seq1 and Seq3 on the expression of matrix metalloproteinase (MMP)-1 and MMP-9 in UVA-induced HaCat cells. (I) Relative MMP-1 and MMP-9 messenger RNA (mRNA) levels in Seq1 and Seq3 treated UVA-induced HaCat cells. Gene expression was normalized to glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ). (J) Western blot analysis showing the change of inducible nitric oxide synthase (iNOS) and interleukin-1 beta (IL-1β) in HaCat cells. (K) Quantitation of IL-1β and tumor necrosis factor-alpha ( TNF-α ) released by HaCat cells by quantitative real-time polymerase chain reaction (qRT-PCR). (L) Glutathione peroxidase (GSH-Px) activity levels in HaCaT cells. (M) Measurement of superoxide dismutase (SOD) activity levels in HaCaT cells. (N) Nrf2-dependent antioxidant enzymes protein levels for reduced nicotinamide adenine dinucleotide phosphate (NAD(P)H) quinone oxidoreductase 1 (NQO1), heme oxygenase 1 (HO-1), and glutamate-cysteine ligase modifier subunit (GCLM) in UVA-irradiated HaCaT cells treated with peptides. Protein expression normalized to GAPDH. (O) Quantitation of GCLM , HO-1 , and NQO1 released by HaCat cells by qRT-PCR. Unless otherwise indicated in the figure, the peptide concentration was 20 μM. All data are presented as means ± standard deviation (SD) ( n = 3). Statistical significance is denoted by ∗ P < 0.05 , ∗∗ P < 0.01 , ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001 vs. UVA-irradiated group.

Article Snippet: Levels of IL-1β and IL-6 in the serum were determined using the Mouse IL-1β enzyme-linked immunosorbent assay (ELISA) Kit (E-EL-M0037, Elabscience, Wuhan, China) and Mouse IL-6 ELISA Kit (E-EL-M0044, Elabscience, Wuhan, China).

Techniques: Migration, Activity Assay, Quantitation Assay, Immunofluorescence, Irradiation, Staining, Fluorescence, Western Blot, Expressing, Gene Expression, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Concentration Assay, Standard Deviation

Journal: Journal of Pharmaceutical Analysis

Article Title: Novel bioactive peptides targeting Keap1-Nrf2 interaction for combating UVA-induced skin aging: Computational discovery and experimental validation

doi: 10.1016/j.jpha.2025.101446

Figure Lengend Snippet: Seq1 and Seq3 mitigated photoaging of fibroblasts (Fb) in vitro . (A) Representative immunofluorescence (IF) staining images of positive cells of reactive oxygen species (ROS) (green) and 4,6-diamidino-2-phenylindole (DAPI). (B) Fluorescence intensity of ROS levels. (C) Representative IF staining images of positive cells of γ-H2AX (green) and DAPI. (D) Quantitation of the mean number of γ-H2AX foci/cell. (E) Quantitation of interleukin ( IL ) -6 , IL-1β , and tumor necrosis factor alpha ( TNF-α ) released by Fb was detected through quantitative real-time polymerase chain reaction (qRT-PCR). (F) Representative IF pictures of unclear factor erythroid 2-related factor (Nrf2) in control and Seq1 or Seq3-treated Fb. (G) Quantification of messenger RNA (mRNA) expression for Nrf2 from Seq1 or Seq3-treated Fb. (H) Quantitation of glutamate-cysteine ligase modifier subunit ( GCLM ), heme oxygenase 1 ( HO-1 ), and nicotinamide adenine dinucleotide phosphate (NAD(P)H) quinone oxidoreductase 1 ( NQO1 ) released by Fb was detected through qRT-PCR. (I) Expression levels of matrix metalloproteinase (MMP)-1, MMP-9, collagen type I alpha 2 (COL1A2), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) measured by Western blot analysis. (J) Quantitation of collagen type III alpha 1 ( COL3A1 ) released by Fb was detected through qRT-PCR. (K) Quantitation of MMP-1 and MMP-9 released by Fb was detected through qRT-PCR. All data are presented as means ± standard deviation (SD) ( n = 3). Statistical significance is denoted by ∗ P < 0.05 , ∗∗ P < 0.01 , ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001 vs. ultraviolet A (UVA)-irradiated group.

Article Snippet: Levels of IL-1β and IL-6 in the serum were determined using the Mouse IL-1β enzyme-linked immunosorbent assay (ELISA) Kit (E-EL-M0037, Elabscience, Wuhan, China) and Mouse IL-6 ELISA Kit (E-EL-M0044, Elabscience, Wuhan, China).

Techniques: In Vitro, Immunofluorescence, Staining, Fluorescence, Quantitation Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Control, Expressing, Western Blot, Standard Deviation, Irradiation

Journal: Journal of Pharmaceutical Analysis

Article Title: Novel bioactive peptides targeting Keap1-Nrf2 interaction for combating UVA-induced skin aging: Computational discovery and experimental validation

doi: 10.1016/j.jpha.2025.101446

Figure Lengend Snippet: The mitigation of ultraviolet A (UVA)-induced skin damage in mice by Seq1 and Seq3. (A) Diagrammatic representation of the mouse photoaging model setup, drawn by Figdraw. (B) Weekly changes in the body weight of mice after UVA irradiation. (C, D) Histological assessment via representative hematoxylin and eosin (H&E) (C) and Masson’s Trichrome (D) staining of mouse skin tissue. Adjacent graphs depict quantitative analysis of epidermal thickness and collagen density ratios. (E) Immunohistochemical detection of the peptides’ effects on UV-treated mouse skin using paraffin-embedded sections. (F) Histograms illustrate enzyme-linked immunosorbent assay (ELISA)-determined serum concentrations of interleukin (IL)-6 and IL-1β for each group. (G) Tyrosinase (TYR) activity was quantified in dorsal skin samples from all groups after UVA or mock-UVA exposure. (H) Hydroxyproline (HYP) content was quantified in dorsal skin samples from all groups after UVA or mock-UVA exposure. All data are presented as means ± standard deviation (SD) ( n = 3). Statistical significance is denoted by ∗∗ P < 0.01 , ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001 vs. UVA-irradiated group. ATRA: all-trans retinoic acid; COL1A2: collagen type I alpha 2; MMP: metalloproteinase.

Article Snippet: Levels of IL-1β and IL-6 in the serum were determined using the Mouse IL-1β enzyme-linked immunosorbent assay (ELISA) Kit (E-EL-M0037, Elabscience, Wuhan, China) and Mouse IL-6 ELISA Kit (E-EL-M0044, Elabscience, Wuhan, China).

Techniques: Irradiation, Staining, Immunohistochemical staining, Enzyme-linked Immunosorbent Assay, Activity Assay, Standard Deviation

Journal: International Journal of Cardiology. Cardiovascular Risk and Prevention

Article Title: The sphingolipid metabolite sphingosine protects against hypertension by targeting metabolic-inflammatory crosstalk via the NLRP3 inflammasome

doi: 10.1016/j.ijcrp.2025.200562

Figure Lengend Snippet: SPH attenuates pathological changes and suppresses NLRP3 inflammasome activation in the hearts of hypertensive mice. (A, B) Masson's trichrome staining of heart (A) and aorta (B) sections, showing reduced collagen deposition (blue) with SPH treatment. Scale bars: 20 μm for heart, 50 μm (main) and 20 μm (inset) for aorta. (C) H&E staining of heart sections showing amelioration of myocardial disarray and inflammation. Scale bars: 500 μm (main) and 20 μm (inset).(D) ELISA quantification of serum IL-18 and IL-1β levels. Data are mean ± SEM. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001 vs. HBP group. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Serum levels of interleukin-18 (IL-18) and interleukin-1β (IL-1β) were quantified using commercial enzyme-linked immunosorbent assay (ELISA) kits (IL-18: Lianke, Cat#EK218-96; IL-1β: Lianke, Cat# EK201BHS-96) according to the manufacturers' instructions.

Techniques: Activation Assay, Staining, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Cardiology. Cardiovascular Risk and Prevention

Article Title: The sphingolipid metabolite sphingosine protects against hypertension by targeting metabolic-inflammatory crosstalk via the NLRP3 inflammasome

doi: 10.1016/j.ijcrp.2025.200562

Figure Lengend Snippet: SPH mitigates Angiotensin II-induced inflammasome activation, apoptosis, and oxidative stress in HUVECs. (A, B) Representative immunofluorescence images showing increased expression of NLRP3 (red, A) and ASC (green, B) in AngII-treated cells, which is reduced by subsequent SPH treatment. Scale bar = 100 μm. (C) TUNEL assay (green) demonstrating increased apoptosis in AngII-treated cells, which is attenuated by SPH. Scale bar = 100 μm. (D, E) Biochemical assays showing that SPH or MCC950 treatment reverses the AngII-induced decrease in SOD1 activity (G).NO (H) and increase in MDA levels (D). (F, G) DCFH-DA staining showing increased intracellular ROS (green) after AngII treatment, which is suppressed by SPH or MCC950. Representative images (F) and quantification (E) are shown. (I, J) ELISA results showing that SPH or MCC950 treatment inhibits the AngII-induced secretion of IL-1β (I) and IL-18 (J). (K) Scanning electron microscopy (SEM) images showing that SPH or MCC950 treatment improves the cell surface morphology and reduces features of pyroptotic damage induced by AngII. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Serum levels of interleukin-18 (IL-18) and interleukin-1β (IL-1β) were quantified using commercial enzyme-linked immunosorbent assay (ELISA) kits (IL-18: Lianke, Cat#EK218-96; IL-1β: Lianke, Cat# EK201BHS-96) according to the manufacturers' instructions.

Techniques: Activation Assay, Immunofluorescence, Expressing, TUNEL Assay, Activity Assay, Staining, Enzyme-linked Immunosorbent Assay, Electron Microscopy

Journal: Molecular Neurobiology

Article Title: Sustained Transcriptional Response to Lipopolysaccharide and Interleukin-4 in an Immortalized Mouse Microglial Cell Line

doi: 10.1007/s12035-026-05711-4

Figure Lengend Snippet: Transcriptomic profiles of LPS and IL-4-specific responders and RNA-seq validation results . A , B Volcano plots showing specific sustained DEGs after A 100 ng/mL LPS and B 25 ng/mL IL-4 treatments, excluding common responder genes. Downregulated genes are shown in blue and upregulated genes in red. Specific responder genes used for validation are labeled. C qPCR validation of RNA-seq data. Transcript levels of Igf1 , Mrc1, Nfkbia , and Ptgs2 after 100 ng/mL LPS, and Igf1 and Il1b after 25 ng/mL IL-4 were measured at 12 h and 24 h. H3f3a was used as a reference gene to normalize the fold changes (2 –ΔΔCt ) relative to the control conditions (dotted line). Data are shown as mean ± SEM ( n ≥ 3). Statistical significance was determined using the mixed-effects model followed by Dunnett’s multiple comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control. D , E Levels of (D) IL-1β and E NO in culture supernatants after 24 h of treatment, measured using the Griess assay and ELISA, respectively. Data are presented as mean ± SEM ( n ≥ 3). Statistical significance was determined by two-way ANOVA followed by Dunnett’s multiple comparisons test. *** p < 0.001 vs. control

Article Snippet: IL-1β levels in culture supernatants were quantified by using a mouse IL-1β enzyme-linked immunosorbent assay (ELISA) kit (#KE10003; Proteintech, Rosemont, IL, USA).

Techniques: RNA Sequencing, Biomarker Discovery, Labeling, Control, Griess Assay, Enzyme-linked Immunosorbent Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Dual metabolic-inflammation modulation in MicroRNA@neutrophil-derived microvesicles achieve robust osteoarthritis therapy

doi: 10.1016/j.apsb.2025.09.020

Figure Lengend Snippet: miR140@MVs inhibit the pro-inflammatory effect of LPS-induced BMDM in vitro . (A) The intracellular location of Cy3-miR140@FITC-MVs in BMDM cells stimulated with LPS (100 ng/mL) for 24 h (M1 macrophages). miR140 is labeled with Cy3 (Red) and MVs are stained by FITC-Annexin V (Green). The nuclei were stained by Hoechst 33342 (Blue). Scale bar: 5 μm. (B) Representative flow cytometric analysis of BMDM treated with different formulations. The (C) M1 (CD86 + CD206 – ) and (D) M2 (CD86 − CD206 + ) phenotypes were analyzed by Flowjo. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, compared with M1 group. n = 3. No significance, ns; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗∗ P < 0.0001. (E) Western blotting of TLR4 in LPS-induced BMDM treated with different formulations. (F) The relative expression of the protein band was analyzed by Image J and normalized to its respective baseline controls. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. ∗ P < 0.05. The relative mRNA levels of (G) TNFα , (H) IL6 , (I) IL1β , (J) IL10 , (K) TGFβ in LPS-induced BMDM receiving different formulations. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, compared with the M1 group, n = 3. No significance, ns; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. (L) The TNF α level in LPS-induced BMDM supernatant was detected by ELISA assay. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. No significance, ns; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. (F–I) Values are normalized to the M1 (baseline control) group.

Article Snippet: Cytokine concentration in the supernatant was quantified by mouse TNF α or mouse IL1 β enzyme-linked immunosorbent assay (ELISA) kits (Elabscience, USA).

Techniques: In Vitro, Labeling, Staining, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Dual metabolic-inflammation modulation in MicroRNA@neutrophil-derived microvesicles achieve robust osteoarthritis therapy

doi: 10.1016/j.apsb.2025.09.020

Figure Lengend Snippet: Single miR140@MVs for prolonged OA treatment. (A) Schematic illustration of the administration regimen. (B) The relative miR140 level in the whole-knee joint of DMM mice treated with different formulations for 28 days. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. The normal group served as the control. No significance, ns. (C) Typical images of imageological examinations, histological analysis, TUNEL staining and immunohistochemical analysis. Red arrows: synovial inflammation; Yellow arrows: cartilage defect; Blue arrows: bone marrow edema; Red circles: periarticular osteophytes. M: meniscus; S: synovial; C: cartilage. Scale bars in H&E and safranine O-fast green staining: 100 μm; in TUNEL staining and immunohistochemical analysis: 50 μm. (D) Heatmap of variables of histological scoring in each group. (E) OARSI grades of the mice joints in each group. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. The normal group served as the control. No significance, ns; ∗∗∗∗ P < 0.0001. (F–J) The relative mRNA levels of inflammatory factors in OA joints from mice after different treatments, including (F) TNFα , (G) IL6 , (H) IL1β , (I) IL10 and (J) TGFβ . The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. The normal group served as control. No significance, ns; ∗∗ P < 0.01; ∗∗∗∗ P < 0.0001.

Article Snippet: Cytokine concentration in the supernatant was quantified by mouse TNF α or mouse IL1 β enzyme-linked immunosorbent assay (ELISA) kits (Elabscience, USA).

Techniques: Control, TUNEL Assay, Staining, Immunohistochemical staining