Compositions containing monoacetyldiacylglycerol compound as an active ingredient for preventing or treating rheumatoid arthritis
a technology of monoacetyldiacylglycerol and monoacetyldiacylglycerol, which is applied in the direction of biocide, drug composition, immunodeficiency disorder, etc., can solve the problems of limited treatment, ineffective treatment methods, and inability to prevent rheumatoid arthritis, so as to prevent, treat or improve rheumatoid arthritis, inhibit the effect of phosphorylation
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[0043]The following examples are provided for better understanding of this invention. However, the present invention is not limited by the examples.
experimental example 1
Inhibitory Effect of EC-18 on STAT-3 Phosphorylation in U937 Cells
[0045]Cells treated in EC-18 and IL-6 cytokines were lysed with cold SDS lysis buffer [(50 mM HEPES, 150 mM NaCl, 0.2 mM EDTA, 0.5% NP-40, 0.1% SDS, 1 mM Na3VO4, 10 mM NaF, and complete Protein Inhibitor Cocktail (Roche)] for 30 minutes on ice. After cell lysis, the aqueous solution was separated from the insoluble precipitate by centrifuging the cell lysates for 30 minutes at 13,000 rpm in a high-speed centrifuge. After protein quantification, the aqueous solution was separated in 10 to 12% SDS-PAGE by electrophoresis. Proteins separated in the gel were transferred into PVDF membrane (Millipore, Billerica, Mass., USA) at 100V for 2 hours.
[0046]To measure the amount of phosphorylated STAT-3, the membrane was incubated with poly rabbit-anti-(STAT1, STAT3) or poly rabbit-anti-phospho (STAT1, STAT3) antibody (Cell Signaling Technology, USA (1:1000) as a primary antibody for 60 minutes at room temperature. The membrane wa...
experimental example 2
Inhibitory Effect of EC-18 on STAT-3 Phosphorylation in NK-92 Cells
[0048]Cells treated in EC-18 and PMA (Phobol 12-myristate 13-acetate) were lysed with cold SDS lysis buffer [(50 mM HEPES, 150 mM NaCl, 0.2 mM EDTA, 0.5% NP-40, 0.1% SDS, 1 mM Na3VO4, 10 mM NaF, and complete Protein Inhibitor Cocktail (Roche)] for 30 minutes on ice. After cell lysis, the aqueous solution was separated from the insoluble precipitate by centrifuging the cell lysates for 30 minutes at 13,000 rpm in a high-speed centrifuge. After protein quantification, the aqueous solution was separated in 10 to 12% SDS-PAGE by electrophoresis. Proteins separated in the gel were transferred into PVDF membrane (Millipore, Billerica, Mass., USA) at 100V for 2 hours.
[0049]To measure the amount of phosphorylated STAT-3 and STAT-1, the membrane was incubated with poly rabbit-anti-STAT3, poly rabbit-anti-STAT1, or poly rabbit-anti-phospho-STAT3 antibody (Cell Signaling Technology, USA (1:1000) as a primary antibody for 60 min...
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