Effect Of Curcumin On Lectin-like Oxidized Low Density Lipoprotein Receptor 1(LOX-1)mRNA Expression In Cardiomyocytes Induced By Angiotensin Ⅱ

Objective: We cultured neonatal SD rat cardiomyocytes,treated them with solvents or drug(curcumin),anti-LOX-1 antibody,and then stimulated with AngⅡ.Observed the difference of the mRNA expression of Lectin-like oxidized low density lipoprotein receptor 1 (LOX-1) among the four groups .To establish the effects of curcumin on the mRNA expression of LOX-1 in cardiomyocytes cultured cells,and further investigate this effect of protecting and delaying some myocardial diseases.Methods: The experiment was accomplished between 2010/3-2010/9 in the the animal operating room of Anhui Medical Univeisity,and the center laboratory of the First Affiliated Hospital of Anhui Medical University.①N eonatal SD rat 6-10(1-2 days),take the hearts out,neonatal rat heart tissue was digested 4-5 times,and then cultured with DMEM/F12 including 10% BSA.②The cardiomyocytes were cultured for 48 hours in DMEM/F12 including 10% BSA,and then changed the culture medium to DMEM/F12 without BSA,cultured for 24 hours.③The cultured cardiomyocytes were divided into four groups:group A,the negative control group;group B,the Positive control group(treated with AngⅡ), group C,the AngⅡ+anti-LOX-1 antibody group,group D,the AngⅡ+curcumin group.④C ultured with DMEM/F12 without BSA for 24 hours,changed the group C's culture medium to DMEM/F12 including 10μg/ml anti-LOX-1 antibody and the group D's culture medium to DMEM/F12 including 5μmol/L curcumin for 3 hours.Then changed the B,C,D group's culture medium to DMEM/F12 including 0.1μmol/L AngⅡ, cultured for another 24 hours.⑤Collected the total RNA of the cultured cardiomyocytes with Trizol,used RT-PCR to detecte the mRNA expression of LOX-1 in four groups,the results were treated with gel-imaging system.Reasults:①This experiment was repeated for three times,the cultured cardiomyocytes growed well.The survival cardiomyocytes was assessed by 0.4% typsin,the survival rate was between 95%-97%,70%-80% beating cardiomyocytes were observed and the beating frequency of cells was 8-70 beats/min.In the area which the cardiomyocytes density was higher,the beating frequency was accordance between 30-70 beats/min.②L OX-1 mRNA expression induced by angiotensinⅡwas obviously upregulated than the negative control group (p<0.05).③Pretreatment of cells with curcumin reduce the increase of LOX-1 mRNA expression treated with AngⅡ(p<0.05),and the expression was not obviously different compaired with pretreatment of cells with anti-LOX-1 antibody(p>0.05).Conclusion: Curcumin can anti-oxidation and change the ventricular remodeling by down-regulate the mRNA expression of LOX-1 induced by AngⅡ.