Journal: Chinese Medicine

Article Title: Matrine improves bile acid metabolism and reduces inflammatory and oxidative stress in colitis via the JAK2 pathway

doi: 10.1186/s13020-026-01387-z

Figure Lengend Snippet: Effects of matrine on intestinal pathology, inflammatory function, and JAK2 expression in experimental colitis mice Dextran Sulfate Sodium (DSS)-induced murine model of colitis was established in mice; matrine and 5-ASA treatment was administered as described. A – C Body weight and colon length were measured. D , E Histopathological alterations in colon tissues were evaluated using H & E staining; DAI scores were assigned according to the staining. F , G The levels of IL-1β, TNF-α, IL-6, MPO, NO, and MDA in colon tissues were examined using commercial assay kits. H , I The protein levels of JAK2 in mice colon tissues were determined using Immunohistochemical staining (IHC staining) and Immunoblotting. Magnification = 100 × or 200 × for IHC staining. J The protein levels of p-JAK2, p-STAT3, and STAT3 were detected using Immunoblotting. **p < 0.01, vs. PBS group; #p < 0.05, ##p < 0.01 vs. DSS group

Article Snippet: Membrane was blocked within Odyssey blocking buffer (LI-COR Bioscience, Lincoln, USA) for 1 h at room temperature (RT), and then incubated with primary antibodies against p-JAK2 (AP0917, Abclonal, Woburn, USA), JAK2 (AF6022, Affinitiy Bioscience, Changzhou, China), MRP3 (bs-0656R, Bioss, Beijing, China), MRP4 (DF6921, Affinity Bioscience), IL-1β [12242, Cell Signaling Technology (CST), Danvers, USA], TNF-α (11948, CST), IL-6 (CSB-PA06757A0RB, CUSABIO, Wuhan, China), p-STAT3 (AF3293, Affinity Bioscience), STAT3 (10253-2-AP, Proteintech, Wuhan, China), p-STAT1 (28977-1-AP, Protientech), STAT1 (10144-2-AP, Proteintech), FXR (M022312, Abmart, Shanghai, China), and GAPDH (endogenous control, 60004-1-Ig, Proteintech) overnight at 4 °C (dilution 1:1000).

Techniques: Expressing, Staining, Immunohistochemical staining, Immunohistochemistry, Western Blot

Journal: Chinese Medicine

Article Title: Matrine improves bile acid metabolism and reduces inflammatory and oxidative stress in colitis via the JAK2 pathway

doi: 10.1186/s13020-026-01387-z

Figure Lengend Snippet: Effects of JAK2 overexpression on intestinal epithelial cells upon inflammation MODE-K cells were transfected with a JAK2-overexpressing plasmid (JAK2 oe) or a control vector, with or without subsequent LPS stimulation. A The protein levels of JAK2 and p-JAK2 and the inflammatory factors IL-1β, TNF-α, and IL-6 were determined by Immunoblotting. B Cellular levels of MPO, NO, and MDA were measured using biochemical kits. C Intracellular ROS production was assessed by DCFH-DA staining and quantified with flow cytometry. D The protein levels of the bile acid metabolism-related factors FXR, MRP3 and MRP4 were examined by Immunoblotting. E The mRNA levels of MRP3, MRP4, OSTα, and OSTβ were measured by qRT-PCR. **p < 0.01 vs. Vector group; ##p < 0.01 vs. Vector group; &&p < 0.01 vs. LPS + Vector group

Article Snippet: Membrane was blocked within Odyssey blocking buffer (LI-COR Bioscience, Lincoln, USA) for 1 h at room temperature (RT), and then incubated with primary antibodies against p-JAK2 (AP0917, Abclonal, Woburn, USA), JAK2 (AF6022, Affinitiy Bioscience, Changzhou, China), MRP3 (bs-0656R, Bioss, Beijing, China), MRP4 (DF6921, Affinity Bioscience), IL-1β [12242, Cell Signaling Technology (CST), Danvers, USA], TNF-α (11948, CST), IL-6 (CSB-PA06757A0RB, CUSABIO, Wuhan, China), p-STAT3 (AF3293, Affinity Bioscience), STAT3 (10253-2-AP, Proteintech, Wuhan, China), p-STAT1 (28977-1-AP, Protientech), STAT1 (10144-2-AP, Proteintech), FXR (M022312, Abmart, Shanghai, China), and GAPDH (endogenous control, 60004-1-Ig, Proteintech) overnight at 4 °C (dilution 1:1000).

Techniques: Over Expression, Transfection, Plasmid Preparation, Control, Western Blot, Staining, Flow Cytometry, Quantitative RT-PCR

Journal: Chinese Medicine

Article Title: Matrine improves bile acid metabolism and reduces inflammatory and oxidative stress in colitis via the JAK2 pathway

doi: 10.1186/s13020-026-01387-z

Figure Lengend Snippet: Effects of matrine on intestinal epithelial cell function upon inflammation Mouse intestinal epithelial cell line, MODE-K, was stimulated with LPS with or without matrine treatment (1, 2, 3 mg/ml), and examined for the protein levels of IL-1β, TNF-α, and IL-6 using Immunoblotting ( A ); The levels of MPO, NO, and MDA using commercial kits ( B ); Intracellular ROS levels were detected by flow cytometry ( C ); The protein levels of FXR, MRP3 and MRP4 were examined using Immunoblotting ( D ); The mRNA levels of MRP3, MRP4, OSTα and OSTβ were examined using qRT-PCR ( E ); The protein levels of p-JAK2, JAK2, p-STAT3, STAT3, p-STAT1, and STAT1 using Immunoblotting ( F ). **p < 0.01, vs. PBS group; #p < 0.05, ##p < 0.01 vs. LPS group

Article Snippet: Membrane was blocked within Odyssey blocking buffer (LI-COR Bioscience, Lincoln, USA) for 1 h at room temperature (RT), and then incubated with primary antibodies against p-JAK2 (AP0917, Abclonal, Woburn, USA), JAK2 (AF6022, Affinitiy Bioscience, Changzhou, China), MRP3 (bs-0656R, Bioss, Beijing, China), MRP4 (DF6921, Affinity Bioscience), IL-1β [12242, Cell Signaling Technology (CST), Danvers, USA], TNF-α (11948, CST), IL-6 (CSB-PA06757A0RB, CUSABIO, Wuhan, China), p-STAT3 (AF3293, Affinity Bioscience), STAT3 (10253-2-AP, Proteintech, Wuhan, China), p-STAT1 (28977-1-AP, Protientech), STAT1 (10144-2-AP, Proteintech), FXR (M022312, Abmart, Shanghai, China), and GAPDH (endogenous control, 60004-1-Ig, Proteintech) overnight at 4 °C (dilution 1:1000).

Techniques: Cell Function Assay, Western Blot, Flow Cytometry, Quantitative RT-PCR

Journal: Chinese Medicine

Article Title: Matrine improves bile acid metabolism and reduces inflammatory and oxidative stress in colitis via the JAK2 pathway

doi: 10.1186/s13020-026-01387-z

Figure Lengend Snippet: Dynamic effects of matrine and JAK2 on intestinal epithelial cell function upon inflammation MODE-K cells were stimulated with LPS with or without matrine treatment (2 mg/ml), transfected with JAK2-overexpressing plasmid (JAK2 oe), and examined for the protein levels of IL-1β, TNF-α, and IL-6 using Immunoblotting ( A ); the levels of MPO, NO, and MDA using commercial kits ( B ); Intracellular ROS levels were detected by flow cytometry ( C ); The protein levels of FXR, MRP3 and MRP4 were examined using Immunoblotting ( D ); The mRNA levels of MRP3, MRP4, OSTα and OSTβ were examined using qRT-PCR ( E ); the protein levels of JAK2, p-STAT3, STAT3, p-STAT1, and STAT1 using Immunoblotting ( F ). **p < 0.01, vs. LPS group; #p < 0.05, ## p < 0.01 vs. LPS + matrine + vector group

Article Snippet: Membrane was blocked within Odyssey blocking buffer (LI-COR Bioscience, Lincoln, USA) for 1 h at room temperature (RT), and then incubated with primary antibodies against p-JAK2 (AP0917, Abclonal, Woburn, USA), JAK2 (AF6022, Affinitiy Bioscience, Changzhou, China), MRP3 (bs-0656R, Bioss, Beijing, China), MRP4 (DF6921, Affinity Bioscience), IL-1β [12242, Cell Signaling Technology (CST), Danvers, USA], TNF-α (11948, CST), IL-6 (CSB-PA06757A0RB, CUSABIO, Wuhan, China), p-STAT3 (AF3293, Affinity Bioscience), STAT3 (10253-2-AP, Proteintech, Wuhan, China), p-STAT1 (28977-1-AP, Protientech), STAT1 (10144-2-AP, Proteintech), FXR (M022312, Abmart, Shanghai, China), and GAPDH (endogenous control, 60004-1-Ig, Proteintech) overnight at 4 °C (dilution 1:1000).

Techniques: Cell Function Assay, Transfection, Plasmid Preparation, Western Blot, Flow Cytometry, Quantitative RT-PCR

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: The relationship between ERβ/IL6 expression levels and clinical characteristics of NSCLC patients

Article Snippet: Human anti-IL6 (GeneTex, San Antonio, Texas, USA).

Techniques: Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: List of biopsies included in this study with patient clinicopathologic parameters and immunohistochemical staining scores for ERβ and IL6, the relationship of ERβ/IL6 expression lever between primary tumor and metastatic lymph nodes

Article Snippet: Human anti-IL6 (GeneTex, San Antonio, Texas, USA).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: IL6 is upregulated in lung cancer and correlates with ERβ expression. ( a ) Immunohistochemical staining of IL6 and ERβ protein in human lung cancer tissue. The brown color in cancer cells denotes positive staining. A negative control was performed without primary antibody. Representative images of IL6 and ERβ expression in NSCLC are shown, including both squamous carcinoma and adenocarcinoma. Original magnification × 100 and × 400. Scale bars 50 and 20 μm respectively. ( b ) The relative level of IL6 expression was plotted against the relative level of ERβ expression in 289 samples (r (spearman) =0.263; p = 0.004). ( c ) Primary NSCLC tumor tissues and metastatic lymph node tissues in three typical cases (original magnification 100× (left) and 400× (right)). ( d ) IHC scores of 30 pairs of specimens from both primary tumor tissue and metastatic lymph nodes (□: lymph nodes metastasis; ○: primary tumor). ( e ) Western blotting analysis of IL6 expression in three paired primary NSCLC tissues and matched metastatic lymph node tissues. GAPDH was used as a loading control

Article Snippet: Human anti-IL6 (GeneTex, San Antonio, Texas, USA).

Techniques: Expressing, Immunohistochemical staining, Staining, Negative Control, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: The relationship between ERβ and IL6 expression levels of 289 NSCLC tissue

Article Snippet: Human anti-IL6 (GeneTex, San Antonio, Texas, USA).

Techniques: Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: Upregulation of IL6 stimulated by E2 in NSCLC cell lines, A549 and H975. ( a ) ERβ/IL6 expression was up regulated in NSCLC cell lines compared with normal pneumocytes (HBE). ( b ) Synchronized cells were treated with E2 at different concentrations (0 nM, 1 nM, 10 nM and 100 nM) and Ful (0.1 μM and 1 μM) for 2 days. The protein expression of ERβ and IL6 was analyzed using western blot. ( c ) Synchronized cells were treated with E2 at different time points (0 h, 6 h, 12 h, 24 h, 48 h and 72 h) at concentrations of 10 nM. The protein expression of ERβ and IL6 was analyzed using western blot. GAPDH was used as a control. ( d ) Autocrine IL6 was analyzed by ELISA assay after concentration-dependent treatment of E2 (10 nM) or Ful (0.1uM). ( e ) The upregulation of IL6 in A549 after stimulation with E2 (10 nM) or Ful (0.1uM) was determined by immunofluorescence

Article Snippet: Human anti-IL6 (GeneTex, San Antonio, Texas, USA).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay, Immunofluorescence

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: Upregulation of IL6 stimulated by E2 treatment regulates aggressiveness of NSCLC cells. ( a ) The cell proliferation of A549 was determined by CCK-8 assay after 72 h concentration-dependent treatment of E2 (10 nM) or Ful (0.1uM) or rhIL6 (0.5 ng/ml) or Stat3 inhibitor static (20uM). The results are representative of three independent experiments. ( b ) Colony formation assay measuring the proliferative activity in A549 cells after concentration-dependent treatment of E2 (10 nM) or Ful (0.1uM) or rhIL6 (0.5 ng/ml) or Stat3 inhibitor static (20uM). ( c ) Wound-healing assays were performed to assess NSCLC cell migration. Wound closure was determined 24 h after the scratch. ( d ) Transwell assay was used to quantify cell migration and invasion capacity. The average number of cells per field of view in three different experiments is plotted. ( e - f ) ELISA and western blot were used to detect the effect of E2 (10 nM) and its receptor antagonist Ful (0.1uM) on IL6 expression and the influence of MEK inhibitor U0126 (60 nM) or a selective PI3K inhibitor of LY294002 (0.6uM) on E2-mediated IL6 expression through MEK/ERK and PI3K/AKT activation in A549 cells

Article Snippet: Human anti-IL6 (GeneTex, San Antonio, Texas, USA).

Techniques: CCK-8 Assay, Concentration Assay, Colony Assay, Activity Assay, Migration, Transwell Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Activation Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: E2 regulates IL6 expression through ERβ and affects malignancy of lung cancer. ( a ) Autocrine IL6 was analyzed by ELISA assay after overexpression or knockdown of ERβ. ( b ) Upregulation of IL6 induced by E2 (10 nM) treatment 48 h was determined by immunofluorescence in A549. ( c ) Colony formation assay measuring the proliferative activity in A549 cells after overexpression or knockdown of ERβ. ( d ) Wound-healing assays were performed to assess NSCLC cell migration in response to changes in ERβ expression. ( e - f ) Transwell assay and DQ-collagen invasion assay were used to quantify the effect of ERβ regulation on cell migration and invasion capacity. ( g ) Expression of IL6 by western blotting after modulation of ERβ expression

Article Snippet: Human anti-IL6 (GeneTex, San Antonio, Texas, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Over Expression, Immunofluorescence, Colony Assay, Activity Assay, Migration, Transwell Assay, Invasion Assay, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: Upregulation of IL6 stimulated by E2 induces tumorigenesis in a urethane-induced adenocarcinoma model. ( a ) The process of creating a urethane-induced adenocarcinoma model illustrated in a simplified sequence flow diagram. ( b - d ) Two weeks after urethane was injected, mice were randomly divided into 3 groups ( N = 6/group): E2 (0.09 mg/kg), E2+ Ful (1.46 mg/kg) and blank control. Lungs were removed after 12 weeks of subcutaneous drug treatment. The gross appearance of lung tumor nodes in different groups is indicated by black circles. The tumor numbers of different groups and tumor growth curves were obtained. ( e - f ) IHC staining of ERβ and IL6 shows a significant positive linear correlation between ERβ and IL6 expression. ( g ) Protein expression of ERβ, IL6, p-p38MAPK, p-AKT and p-Stat3 in murine lung tumors was analyzed using western blot. All data are expressed as the mean ± SD. Student’s t-test was used for statistical analysis

Article Snippet: Human anti-IL6 (GeneTex, San Antonio, Texas, USA).

Techniques: Sequencing, Injection, Immunohistochemistry, Expressing, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: ERβ isoforms play differential roles in E2-stimulated IL6 expression. ( a ) The creation of our xenograft mouse model illustrated in a simplified sequence flow diagram. ( b - c ) Ovariectomized mice were subcutaneously injected with A549 cells transfected with shRNA-NC, shRNA-ERβ1, shRNA-ERβ2 or shRNA-ERβ5 shRNA lentiviral particles (GeneChem) ( N = 8/group). Mice were euthanized and photographed after 4 weeks of E2 (0.09 mg/kg) treatment. The tumor photograph, tumor weight and tumor growth curves of each group were obtained. ( d - f ) IHC staining of ERβ, ERβ1 and IL6 showed a significant positive linear correlation between ERβ and IL6 expression. All data are presented as the mean of three independent experiments ± SD. ( g ) The protein expression of ERβ subtypes, IL6, p-p38MAPK, p-AKT and p-Stat3 in murine lung tumors was measured using western blot. All data are expressed as the mean ± SD. Student’s t-test was performed to assess statistical significance

Article Snippet: Human anti-IL6 (GeneTex, San Antonio, Texas, USA).

Techniques: Expressing, Sequencing, Injection, Transfection, shRNA, Immunohistochemistry, Western Blot

Journal: Metabolites

Article Title: Liver Metabolomics and Inflammatory Profiles in Mouse Model of Fentanyl Overdose Treated with Beta-Lactams

doi: 10.3390/metabo13080965

Figure Lengend Snippet: ( A ) IL-6 and CYP3A11 (mouse homolog of human CYP3A4) bands expression in the control, fentanyl, fentanyl–ceftriaxone, and fentanyl-MC100093 groups. ( B ) One-way ANOVA followed by Holm–Sidak’s multiple comparisons test showed that liver IL-6 expression was increased in the fentanyl and fentanyl-MC100093 groups compared to the control and fentanyl–ceftriaxone groups; moreover, liver CYP3A11 expression was lower in the fentanyl and fentanyl-MC-100093 groups compared to the control and fentanyl–ceftriaxone groups ( n = 4/group). Data are reported as mean ± SEM. (* p < 0.05, ** p < 0.01). Cef, ceftriaxone; MC, MC-100093.

Article Snippet: These primary antibodies were rabbit anti-interleukin-6 (IL-6, MyBiosource, San Diego, CA, USA), rabbit anti-CYP3A11 (mouse homolog of human CYP3A4) (ABclonal, Wuhan, China), and rabbit anti-β-actin (ABclonal, Wuhan, China).

Techniques: Expressing