[Establishment of a drug screening model for identifying up-regulator of human high density lipoprotein receptor].
OBJECTIVE
To establish a new drug screening model based on transcriptional regulation of human high density lipoprotein (HDL) receptor gene CD36 and LIMPII analogous-1 (CLA-1) for discovering up-regulator of this receptor.


METHODS
The upstream regulatory sequence of CLA-1 was obtained by polymerase chain reaction. A recombinant reporter plasmid pGL3-CLAP was constructed by inserting the regulatory sequence upstream of luciferase gene of pGL3-Basic. Human hepatoma cell line BEL-7402 was transfected with pGL3-CLAP. Samples were detected by testing luciferase activity of transfected BEL-7402 cells in microtiter wells.


RESULTS
The drug screening model was established and optimized. Significant difference was present between pGL3-CLAP and pGL3-Basic transfected BEL-7402 cells (P< 0.001), and coefficient of variation was less than 10%. After primary and secondary screening, 1 compounds and 3 fermentation extracts had up-regulating activities.


CONCLUSIONS
This new drug screening model may be efficiently used to screen up-regulators of human HDL receptor expression, which might become lead compounds for new anti-atherosclerosis drugs.
Liu
Xiao-hui
XH
Department of Biotechnology, Institute of Medicinal Biotechnology, CAMS and PUMC, Beijing 100050, China.
Hong
Bin
B
Wang
Li-fei
LF
Yang
Yuan
Y
Si
Shu-yi
SY
Li
Yuan
Y
D018955:CD36 Antigens; D002788:Cholesterol Esters / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D000960:Hypolipidemic Agents / Q000138:chemical synthesis* / Q000494:pharmacology; D008075:Lipoproteins, HDL / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins; D011971:Receptors, Immunologic / Q000235:genetics*; D018110:Receptors, Lipoprotein / Q000235:genetics*; D051116:Receptors, Scavenger; D051122:Scavenger Receptors, Class B; D014158:Transcription, Genetic / Q000187:drug effects; D015854:Up-Regulation
D018955:CD36 Antigens; D002788:Cholesterol Esters; D000960:Hypolipidemic Agents; D008075:Lipoproteins, HDL; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011971:Receptors, Immunologic; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; C495124:SCARB1 protein, human; D051122:Scavenger Receptors, Class B; C031616:high density lipoprotein receptors