Journal: British Journal of Pharmacology

Article Title: Dual inhibition of cannabinoid CB 1 receptor and inducible NOS attenuates obesity‐induced chronic kidney disease

doi: 10.1111/bph.14849

Figure Lengend Snippet: MRI‐1867 reverses the fatty acid‐induced reduction in adiponectin signalling. Mice on standard diet (STD) or high‐fat diet (HFD) for 18 weeks were treated with vehicle (Veh) or MRI‐1867 (3 mg·kg −1 ) orally for 28 days. (a–c) MRI‐1867 restored the HFD‐induced reduction in the mRNA renal expression of adiponectin, and Adipo2 but not Adipo1 receptors. Similarly, exposing HK‐2 cells to O:P (0.5 mM, 2:1, respectively) resulted in reduced (d–f) mRNA and protein expression of adiponectin as well as reduced mRNA levels of (g) Adipo1 and (h) Adipo2 receptors. Pretreatment of the cells with MRI‐1867 (100 ng·ml −1 ) completely normalized these changes. (i) A proposed mechanism for the dual blockade of CB 1 receptors and inducible NOS by MRI‐1867 in reversing obesity‐induced chronic kidney disease (CKD) is shown. RQ, relative quantitation. In vivo data represent the mean ± SEM from 8 to 14 mice per group. * P < .05, significantly different from animals on STD; # P < .05, significantly different from animals on the same diet. In vitro data represent the mean ± SEM from five independent experiments. * P < .05, significantly different from Veh‐treated cells under normal conditions; # P < .05, significantly different from Veh‐treated cells under O:P conditions. Data were analysed by one‐way ANOVA, followed by a Bonferroni post hoc test

Article Snippet: HK‐2 cells were transfected with CRISPR‐CAS9 vector containing an sgRNA sequence to target all human isoforms of CNR1 (Genecopeia™, Cat# CS‐HCP263432‐CG01‐01‐B) using Lipofectamine 3000 (Invitrogen, Cat# L3000‐001) and selected with hygromycin (200‐μM; Sigma‐Aldrich, Cat# H3274).

Techniques: Expressing, Quantitation Assay, In Vivo, In Vitro

Journal: Journal of Natural Medicines

Article Title: Allicin ameliorates sepsis-induced acute kidney injury through Nrf2/HO-1 signaling pathway

doi: 10.1007/s11418-023-01745-3

Figure Lengend Snippet: Allicin inhibited the inflammatory response and oxidative stress, and restored mitochondrial function in HK2 cells primed with lipopolysaccharide (LPS). a CCK-8 analysis of the viability of HK2 cells treated with allicin. b CCK-8 analysis of the viability of HK2 cells with LPS and allicin treatments. c Protein levels of IL-6, IL-1β, TNF-α and MCP-1 in HK2 cells with LPS and allicin treatments. d Levels of ROS and MDA, as well as activities of SOD and GSH-Px in LPS-primed HK2 cells after allicin treatment. e Levels of cytochrome c in the mitochondria and cytoplasm of HK2 cells treated with LPS and allicin. f Analysis of JC-1-labeled mitochondrial membrane potential changes by flow cytometry in HK2 cells, and statistical analysis of JC-1 green monomer. g Detection of cell apoptosis by flow cytometry in HK2 cells with LPS and allicin treatment. All data were expressed as mean ± SD. “ns”: no significance, ** p < 0.01, and *** p < 0.001, n = 3 in each group, and the number of technical replicates is 3

Article Snippet: HK2 cell line was purchased from Procell Life Science & Technology (Wuhan, China), and it was cultured in MEM (Solarbio, Beijing, China) supplemented with 10% fetal bovine serum (Tianhang, Zhejiang, China) in an incubator at 37 °C with 5% CO 2 .

Techniques: CCK-8 Assay, Labeling, Membrane, Flow Cytometry

Journal: Journal of Natural Medicines

Article Title: Allicin ameliorates sepsis-induced acute kidney injury through Nrf2/HO-1 signaling pathway

doi: 10.1007/s11418-023-01745-3

Figure Lengend Snippet: Allicin inhibited the cytotoxicity of LPS in HK2 cells by promoting the Nrf2/HO-1 signaling pathway. a Protein levels of cleaved caspase-3, cleaved caspase-9, Bax, and Bcl-2 in HK2 cells after allicin and LPS treatment. b Protein levels of Nrf2 and HO-1 in HK2 cells treated with allicin and LPS. All data were expressed as mean ± SD. c The accumulation of NRF2 in the nucleus was detected by immunofluorescence, bar = 50 μm. * p < 0.05, ** p < 0.01, and *** p < 0.001, n = 3 in each group, and the number of technical replicates is 3

Article Snippet: HK2 cell line was purchased from Procell Life Science & Technology (Wuhan, China), and it was cultured in MEM (Solarbio, Beijing, China) supplemented with 10% fetal bovine serum (Tianhang, Zhejiang, China) in an incubator at 37 °C with 5% CO 2 .

Techniques: Immunofluorescence

Journal: Journal of Natural Medicines

Article Title: Allicin ameliorates sepsis-induced acute kidney injury through Nrf2/HO-1 signaling pathway

doi: 10.1007/s11418-023-01745-3

Figure Lengend Snippet: Validation of the promotive effect of allicin on Nrf2/HO-1 signaling pathway. a Levels of TNF-α, IL-1β and ROS as well as the activity of GSH-Px in HK2 cells with allicin and ML385 treatment. b Analysis of JC-1-labeled mitochondrial membrane potential by flow cytometry in LPS and allicin and ML385 treated HK2 cells. c Evaluation of cell apoptosis by flow cytometry in HK2 cells with LPS and allicin and ML385 treatments. All data were expressed as mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001, it was compared with LPS + allicin-20 group, n = 3 in each group, and the number of technical replicates is 3

Article Snippet: HK2 cell line was purchased from Procell Life Science & Technology (Wuhan, China), and it was cultured in MEM (Solarbio, Beijing, China) supplemented with 10% fetal bovine serum (Tianhang, Zhejiang, China) in an incubator at 37 °C with 5% CO 2 .

Techniques: Biomarker Discovery, Activity Assay, Labeling, Membrane, Flow Cytometry