应用性科学研究讲义.pptVIP

Antagonising of LXR RH150 RH150 is one of main component of Rheum palmatum: has been used to treat constipation in traditional Chinese medicine LXR antagonist – RH150 For this assay, receptor protein is immobilized on the SPR sensor chip, upon ligand binding, the interaction between receptor protein and ligand is enhanced and the kinetic parameters are altered and these changes are monitored in real time. This assay has been proved to be specific, dose dependent and reproducible during the analysis of small molecule compound and protein interaction. SPR(surface plasmon resonance) technology RH150 bind to LXRs directly Figure 1. RH150 exhibits specific binding affinity against LXRs as evaluated by SPR experiments RH150 inhibits the transcriptional activity of LXRs to its agonist GW3965 Figure 2. RH150 inhibits LXR transcriptional activity to its agonist GW3965 through transient transfection luciferase assay on 293T cells RH150 inhibit the LXRs target genes on 3T3-L1 adipocytes and HEPG2 cells Figure 3. RH150 attenuates LXR related genes on 3T3-L1 adipocytes and HEPG2 cells through Real-time PCR after treated with RH150 50uM for 24 hours. In vivo effect on high fat diet-induced obesity RH150 prevents fat accumulation in mice fed with high-fat diet and reduces body Weight and fat weight significantly in diet-induced obese mice Ex vivo confirmation DIO RH150 decreases both hepatic TG, TC levels and serum TC levels and improves glucose tolerance in DIO mice Figure 6. RH150 reduces expression of lipogenic related genes in WAT and liver through real-time PCR. Changed gene expressions involved in lipid and glucose metabolism in DIO mice Figure 7 RH150 elevatesthe expression of energy expenditure related genes and increases the oxygen consumption in DIO mice without change the serum T3,T4 levels. Increased energy expenditure in muscle and BAT in DIO mice RH150 decreases both hepatic TG, TC levels and serum TC levels and i