ABSTRACT
Objective To identify the important regulatory elements in the promoter of human CD2 associated protein(CD2AP) by conserved sequence analysis among different species and luciferase functional detection. Methods The promoter sequences of CD2AP from different species were analyzed by BLAST. Plasmids containing different length of deletion mutations of human CD2AP promoter were constructed. Pro-moter activities were tested in 3 kinds of cells from different species by luciferase analysis and were tested in HEK-293 cells treated with all-trans-retinoic acid. Results Homologous sequence comparison in CD2AP promoter area among human, cattle and pig showed that putative specific protein 1 (Sp1) sites and down-stream promoter element (DPE) were highly evolutional]y conserved. Progressive deletion luciferase analysis of DNA fragments revealed similar promoter activity style among 3 different cell lines from 3 different spe-cies, HEK-293, BHK-21 and Vero cells. One basic promoter activity located within 500 bp upstream of ATG. Fragments of further upstream 100 bp or more had drastically 10 times increased promoter activity. Two putative Sp1 sites were in this 100 bp region. All-trans-retinoic acid decreased the luciferase activity of CD2AP promoter. Conclusion Putative Spl sites and DPE have important functions in the promoter activity of CD2AP.
ABSTRACT
Objective To observe the effect of lovastatin on plasminogen activator inhibitor -1 (PAI-1) and collagen type Ⅳ in rats with glomerularsclerosis induced by adriamycin,and to discuss its mechanism of protective effects on kidneys.Methods Twenty-four male Wistar nephritic rats induced by adriamycin were randomly divided into 3 groups:control,hyperlipidemia and lovastatin treatment group.They were fed 12 weeks.Urinary protein excretion and serum lipid were assayed,then renal glomerularsclerosis index,the expression of PAI-1 and collagen type Ⅳ were observed.Results Serum total cholesterol,triglycerides,low-density lipoprotein,urinary protein excretion,renal glomerularsclerosis index were significantly lower in treatment group than those in hyperlipidemia group.Expression of PAI-1 and collagen type Ⅳ,and number of foamcells were also sharply lower in treatment group than those in hyperlipidemia group.Conclusions Lovastatin not only reduces proteinuria,improves renal function,but also modulates glomerularsclerosis by inhibiting activity of PAI-1 and decreasing accumulation of collagen type Ⅳ.The mechanism of renal protective effect is independent of a reduction of circulating cholesterol.