Effect Of Spironolactone On Lectin-iike Oxidized Low Density Lipoprotein Receptor1(LOX-1) MRNA Expression In HUVECs Induced By Aldosterone

Objective:We cultured human umbilical vein endothelial cells,treated them with spirolactone(Spiro) and the Chemical antagonist of LOX-1polyinosinic acids (POLYI),and then stimulated with Aldosterone (ALD).Observed the difference of the mRNA expression of Lectin-like oxidized low density lipoprotein receptor1(LOX-1) among the six groups.To establish the effects of Spiro on the mRNA expression of LOX-1in human umbilical vein endothelial cells,and further investigate this effect of protecting and delaying some Atherosclerotic diseases.Methods:The experiment was accomplished between2011/5-2011/10in the the center laboratory of the First Affiliated Hospital of Anhui Medical University.①human umbilical vein endothelial cells were purchased from Nanjing KaiJi biological science and technology development Co, LTD and then cultured in1640supplemented with20%(v/v)fetal bovine serum.②The human umbilical vein endothelial cells were cultured for48hours in1640with20%(v/v)fetal bovine serum,,and then changed the culture medium to1640without fetal bovine serum,then Sub-confluent cells having starved in1640containing0.0%FBS for12h.③After12h The cultured human umbilical vein endothelial cells were divided into six groups:the negative control group(group A),treated with different various concentrations of aldosterone group(10nmol/L、100nmol/L、1000nmol/L,group B、C、D),the ALD+polyinosinic acid (the chemical inhibitors of LOX-1) group(group E),the ALD+Spiro group(group F),Group E was pretreated with polyinosinic acid (250mg/L)for2h and then both Group E and group F (treat with Spirolumol/L) stimulated with ALD (1000nmol/l)for24h.④After24h collected the total RNA of the cultured human umbilical vein endothelial cells with TRizol,used RT-PCR to detecte the mRNA expression of LOX-1in six groups,the results were treated with gel-imaging system.Results:①This experiment was repeated for three times,the cultured human umbilical vein endothelial cells growed well.The survival human umbilical vein endothelial cells was assessed by0.4%typsin,the survival rate was between96%-98%,In the area which the human umbilical vein endothelial cells density was higher.②LOX-1mRNA expression induced by Aldosterone was obviously upregulated than the negative control group (p<0.05)There was significant difference.③Aldosterone in a certain concentration range raise the expression of LOX-1mRNA in vitro cultured human umbilical vein endothelial cells and in a dose dependency, the expression of LOX-1mRNA B<C<D, among each other (P<0.05)There was significant difference.④A certain concentration of spironolactone can prevent aldosterone-induced LOX-1mRNA upregulation compared with different concentrations of aldosterone groups (p<0.05).⑤the expression of LOX-1mRNA between the spironolactone group and the negative control group was no significant difference (p>0.05).⑥the expression of LOX-1mRNA of Spironolactone group compared with polyinosinic cytidylic caid group was no significant difference (p>0.05).(D group compared with A group P value was0.001, D group compared with E group and F group, P values were0.001and0.002A group compared with E group and F group,P values were0.584and0.951,E group compared with F group P value was0.544)Conclusion:Spiro May have anti-artery atherosclerosis and anti-oxidation by down-regulate the mRNA expression of LOX-1induced by Aldosterone.