Cannabidiol (CBD): a Killer for Inflammatory Rheumatoid Arthritis Synovial Fibroblasts
In total, 40 patients with long-standing RA fulfilling the American College of Rheumatology revised criteria for RA (24) were included in this study. The RA group comprised of 32 females and 8 males with a mean age of 67.8 years ±10.5 years and 66.9 years ±8.2 years, respectively. C-reactive protein was 47.9 mg/dl ± 186.3 mg/dL for females and 28.7 mg/dL ± 43.2 mg/dL for males and rheumatoid factor was 184.4 iU/mL ± 280.4 iU/mL for females and 31.8 iU/mL ± 37.6 iU/mL for males. IN all, 11 out of 40 patients received glucocorticoids, 7 out of 40 methotrexate, 3 out of 40 biologicals, and 1 out of 40 a JAK inhibitor. All patients underwent elective knee joint replacement surgery, and they were informed about the purpose of the study and gave written consent. Samples from RA synovial tissue were isolated and prepared as described previously22 (for details see also Supplementary methods). G8080) according to manufacturer’s instructions. P6866) for 60 min at 37 °C followed by 30 min at room temperature.
P3584) and respective antagonists/ligands/inhibitors were added for 30 min at room temperature. PoPo3 uptake were evaluated with a TECAN multimode reader over 90 min. RASF were trypsinized, washed and fixed for 20 min with 3.7% formaldehyde (F8775, Sigma Aldrich). Cells were permeabilized with 0.1% Triton X-100 (X100, Sigma) in PBS for 10 min. Then, 0.2 µg/50 µl primary antibody (Proteintech, 19124-1-AP) was added for 2 h. The secondary antibody (Abcam, goat anti-rabbit IgG H&L (Alexa Fluor® 488), ab150077) was incubated for 1 h.
In total, 40 patients with long-standing RA fulfilling the American College of Rheumatology revised criteria for RA (24) were included in this study. The RA group comprised of 32 females and 8 males with a mean age of 67.8 years ±10.5 years and 66.9 years ±8.2 years, respectively. C-reactive protein was 47.9 mg/dl ± 186.3 mg/dL for females and 28.7 mg/dL ± 43.2 mg/dL for males and rheumatoid factor was 184.4 iU/mL ± 280.4 iU/mL for females and 31.8 iU/mL ± 37.6 iU/mL for males. IN all, 11 out of 40 patients received glucocorticoids, 7 out of 40 methotrexate, 3 out of 40 biologicals, and 1 out of 40 a JAK inhibitor. All patients underwent elective knee joint replacement surgery, and they were informed about the purpose of the study and gave written consent. Samples from RA synovial tissue were isolated and prepared as described previously22 (for details see also Supplementary methods). G8080) according to manufacturer’s instructions. P6866) for 60 min at 37 °C followed by 30 min at room temperature.
P3584) and respective antagonists/ligands/inhibitors were added for 30 min at room temperature. PoPo3 uptake were evaluated with a TECAN multimode reader over 90 min. RASF were trypsinized, washed and fixed for 20 min with 3.7% formaldehyde (F8775, Sigma Aldrich). Cells were permeabilized with 0.1% Triton X-100 (X100, Sigma) in PBS for 10 min. Then, 0.2 µg/50 µl primary antibody (Proteintech, 19124-1-AP) was added for 2 h. The secondary antibody (Abcam, goat anti-rabbit IgG H&L (Alexa Fluor® 488), ab150077) was incubated for 1 h.