The In Vitro Effect And Mechanism Studies On Lipid Metabolism Of ABCA1 And CLA-1 UP-Regulators

Objective:The purpose of the study was to find a good leading compound for regulating lipid metabolism and to evaluate the in vitro effect of the active compound on lipid metabolism.Methods:The compound which was ABCA1 and CLA-1 up-regulators was found as a positive hit using cell-based high throughput screening models which were established in Insititute of medicinal biotechnology academy of medical sciences & peking union medical college national center for microbial drug screening. The levels of mRNA and protein were measured by Real-time PCR and Western blot, respectively. The compound of PPAR activation function was found by PPAR nuclear receptor activation experiment. PPAR alpha/gamma antagonists were investigated compound on the mechanisms of gene regulation. The in vitro role of regulating lipid metabolism were reviewed in foam cell assay, cholesterol efflux assay and differentiation of mouse preadipocytes 3T3-L1 assay.Results:E23869 was found as a positive hit using cell-based high throughput screening models which were established in Insititute of medicinal biotechnology academy of medical sciences & peking union medical college national center for microbial drug screening among 20000 compounds screened. The up-regulating activities of E23869 in ABCAlp-LUC and CLA-1 p-LUC HepG2 cells were 196% and 198%, respectively. E23869 could significantly upregulate the mRNA and protein levels of ABCA1, SR-BI/CLA-1 and ABCG1 in both macrophages RAW264.7 and L02 cells by Real-time PCR and Western blot analysis. And also E23869 could not significantly increase the expression of FAS and CD36. Foam cell assay showed that E23869 could inhibit lipids accumulations in macrophages RAW264.7. Cholesterol efflux assay showed that E23869 could induce HDL-mediated cholesterol efflux in macrophages RAW264.7. Moreover, it was found that E23869 up-regulated ABCA1, SR-BI/CLA-1 and ABCG1 expressions through activating PPARα and PPARγ. In addition, E23869 weakly promoted in vitro differentiation of mouse preadipocytes 3T3-L1.Conclusion:As ABCA1 and CLA-1 up-regulators, E23869 could promote cholesterol efflux, which might be a good leading compound for regulating lipid metabolism.