Mouse tnf α elisa kit

Specific pathogen-free male C57Bl/6J mice (weight 18–22 g) were supplied from the Shanghai Laboratory Animal Center, Chinese Academy of Sciences. The animals were housed. All experiments abided by institutional ethical guidelines of the Animal Care and Use Committee (Shanghai Institute of Materia Medica, Chinese Academy of Sciences, China). For drug administration, acetaminophen was dissolved in normal saline, whereas dabrafenib and vemurafenib were dissolved in pH 8.0 distilled water containing 0.5% HPMC and 0.2% Tween 80. Overnight-fasted mice were given dabrafenib (300 mg/kg or 100 mg/kg) or vemurafenib (300 mg/kg) by oral gavage 1 h before the intraperitoneal injection of 300 mg/kg acetaminophen. The mice were killed 6 h post the acetaminophen treatment. Plasma alanine aminotransferase and aspartate aminotransferase were measured with a Cobas C501 Chemistry Analyzer (Roche, Basel, Switzerland). Plasma IL-1β and TNF-α were measured by using the mouse IL-1β ELISA kit and the mouse TNF-α ELISA kit (MultiSciences, Bellingham, WA, USA), respectively. Liver GSSG was measured by using a GSSG Assay kit (Beyotime). To evaluate the changes in liver histology, sections of paraformaldehyde-fixed liver tissues were stained with hematoxylin and eosin and photographed under an optical microscope. The standard TUNEL staining was carried out to evaluate the DNA breaks in liver cells.

TNF-α, IL-1β, IL-10, IL-4, G-CSF and TGF-β1 levels in culture supernatants were measured using a Mouse TNF-α ELISA Kit (MULTI SCIENCES, China), Mouse IL-1β ELISA Kit (MULTI SCIENCES, China), Mouse IL-10 ELISA Kit (Angle Gene, China), Mouse IL-4 ELISA Kit (Angle Gene, China), Mouse G-CSF/CSF3 ELISA Kit (Sigma-Aldrich, USA), and mouse TGF-β1 ELISA kit (KeyGEN BioTech Co., Ltd, China). Moreover, IFN-γ levels in intestinal tissues were measured using a rat IFN-γ ELISA kit (MULTI SCIENCES, China).

After 12 weeks of intervention by Semaglutide, mice were fasted for 12 h, and tail blood was taken to measure fasting blood glucose with a glucose meter (Accu-Chek; Roche Diagnostics GmbH, Germany). Mouse INS (Insulin) ELISA Kit (E-EL-M1382c, Elabscience Biotechnology Co., Ltd, China) was used to determine the level of Insulin. Fasting Insulin Resistance index (HOMA-IR) was also assessed: HOMA-IR=fasting glucose×fasting insulin/22.5. Then, the mice were anesthetized with 40 mg/kg pentobarbital sodium, and the blood from inner canthus was collected and centrifuged at 2000 r/min for 20 min. Finally, the plasma was separated and tested for lipid and inflammatory indexes, including: triglyceride (TG), serum cholesterol (CHO), low-density lipoprotein (LDL), high-density lipoprotein (HDL), tumour necrosis factor-ɑ (TNF-α), interleukin-6 (IL-6), and IL-1β. TG and CHO were detected by the GPO-PAP method (NanJing JianCheng Bioengineering Institute, China). LDL and HDL were measured by the terminal method. Mouse IL-1β ELISA Kit (70-EK201B/3-96, MultiSciences Biotech Co., Ltd., China), Mouse IL-6 ELISA Kit (70-EK206/3-96, MultiSciences Biotech Co., Ltd., China) and Mouse TNF-α ELISA Kit (70-EK282/4-96, MultiSciences Biotech Co., Ltd., China) was used to determine the level of IL-1β, IL-6 and TNF-α, respectively.