LDL receptor gene expression in human mesangial cells under the influence of calcium channel blockers.
BACKGROUND
Intracellular transport of lipid through the regulation of LDL receptor (LDLr) may be important in the progression of renal dysfunction.


METHODS
We explored LDLr gene expression in human mesangial cell line (HMCL) under influence of calcium channel blockers using cell proliferation, LDL binding, Northern blot and LDLr promoter activity assay.


RESULTS
Diltiazem and verapamil increased the expression of LDLr mRNA in a dose-dependent manner. Increased LDLr mRNA paralleled LDL binding. Nifedipine did not increase the expression of LDLr mRNA and LDL binding to HMCL at 1 - 100 micromol/l. The LDLr promoter activity assay showed that treatment with 100 micromol/ of diltiazem and verapamil increased LDLr promoter activity by 126.72 +/- 10.68%, and 166.41 +/- 11.41%, respectively, at 24 hours (control as 100%), while treatment with 100 micromol/l of nifedipine had an inhibitory effect on LDLr promoter activity. High concentration of LDL (250 microg/ml) inhibited promoter activity. Diltiazem or verapamil coincubated with LDL (250 microg/ml) could not override transcriptional inhibition by LDL. CCBs inhibited the proliferation of HMCL, therefore, CCBs-induced LDLr expression did not depend on a proliferative response. Signal transduction pathway experiments showed that the calmodulin transduction pathway was involved in LDLr upregulation induced by diltiazem or verapamil. Additionally, tyrosine kinase and PKC pathways were involved in the induction of LDLr induced by verapamil.


CONCLUSIONS
These studies show that diltiazem and verapamil increase LDLr gene transcription and expression which is independent of cell proliferation in HMCL.
Ruan
X Z
XZ
Centre for Nephrology, Royal Free and University College Medical School, London, UK.
Varghese
Z
Z
Fernando
R
R
Powas
S H
SH
Moorhead
J F
JF
D015152:Blotting, Northern; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004110:Diltiazem / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D005920:Glomerular Mesangium / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism; D009543:Nifedipine / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D011973:Receptors, LDL / Q000235:genetics*; D015398:Signal Transduction / Q000187:drug effects; D014158:Transcription, Genetic / Q000187:drug effects; D014700:Verapamil / Q000494:pharmacology
D002121:Calcium Channel Blockers; D008077:Lipoproteins, LDL; D012333:RNA, Messenger; D011973:Receptors, LDL; D014700:Verapamil; D004110:Diltiazem; D009543:Nifedipine