[Study on the protective effect of HDL subclasses against lipopolysaccharide].
OBJECTIVE
To elucidate the anti-LPS effect of high-density lipoprotein (HDL) subclasses.


METHODS
The pNF-kappaB-luc plasmid was transfected into HepG2 cells and the luciferase expression was obtained by the optimization of cell density, incubation time of transfected cells and stimulating concentration of LPS. The luciferase expression was examined in the cells treated with various stimulus including LPS, mixtures of HDL subclasses and LPS. The expression of TNF-alpha mRNA was detected RT-PCR.


RESULTS
After LPS stimulation, HDL subclasses did not show obvious influence on the effect of LPS to stimulate the luciferase production. However, pre-incubation of cells or pre-incubation of LPS with large-sized HDL(2) strongly inhibited the effect of LPS to stimulate the luciferase production. When LPS was incubated with increased concentration of HDL(2), the LPS effect was decreased to a greater extent. Pre-incubation of LPS with HDL(2) before addition to the cells resulted in a significant decrease in the mRNA level of TNF-alpha.


CONCLUSIONS
The largest HDL(2) has strong LPS-binding capacity and can inhibit the LPS induced TNF-alpha release in HepG2 cells; HDL(3) shows weak anti-LPS effect; small-sized prebeta(1)-HDL does not show anti-LPS effect.
Jia
Lian-qun
LQ
Department of Biochemistry and Molecular Biology, Liaoning University of Traditional Chinese Medicine, Shenyang 110032, China. jlq-8@163.com
Wang
Qi-ming
QM
Liu
Chun
C
Tian
Li
L
Fu
Ming-de
MD
D045744:Cell Line, Tumor; D015870:Gene Expression / Q000187:drug effects*; D053442:High-Density Lipoproteins, Pre-beta / Q000494:pharmacology; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008075:Lipoproteins, HDL / Q000494:pharmacology*; D053439:Lipoproteins, HDL2 / Q000494:pharmacology; D053440:Lipoproteins, HDL3 / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D053442:High-Density Lipoproteins, Pre-beta; D008070:Lipopolysaccharides; D008075:Lipoproteins, HDL; D053439:Lipoproteins, HDL2; D053440:Lipoproteins, HDL3; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D008156:Luciferases