Protection Effect Of AVE3085 On ADMA Induced HCAEC Dysfunction

Objective To study effect of asymmetric dimethylarginine(ADMA) on human coronary artery endothelial cells(HCAEC), and to explore protection effect of eNOS enhancer AVE3085 on ADMA induced HCAEC dysfunction.Methods HCAEC were divided into four groups( Control, ADMA, ADMA+AVE3085,ADMA+DMSO) and curtured 24 h seperately. The expression of eNOS-m RNA was detected by RT-PCR. Protein expressions of eNOS and p-eNOSser1177 were determined by Western Blot. NO and ROS were detected by fluorescence probe. The apoptosis was tested by fluorescence labeling and flow cytometry.Results To compare with the control group, AMDA markly down-regulated the m RNA(F=1041.17,q=80.50,P<0.01), and the protein of eNOS and p-eNOSser1177(F=391.35 ~432.74,q=23.55~34.62,P<0.01). ADMA also inhibited the formation of NO(F=600.36,q=32.18,P<0.01), stimulated the formation of ROS(F=918.05,q=38.62,P<0.01), and induced the apoptosis(F=126.98~361.55,q=13.60~24.53,P<0.01). To compare with the ADMA group, ADMA+AVE3085 may markly restore the eNOS gene(F=1041.17,q=36.83,P<0.01) and protein expression(F=391.35~432.74,q=22.35~35.41,P<0.01). In addition, it also improved the NO-release(F=600.36,q=28.17,P<0.01), inhibited the formation of ROS(F=918.05,q=34.11,P<0.01). The number of apoptotic cells was significantly reduced(F=126.98~361.55,q=12.65~21.91,P<0.01).Conclusion ADMA lead the HCAEC dysfunction. AVE3085 may protect the HCAEC by up-regulating the gene and protein expression, improving the release of NO,inhibiting the fornation of ROS and apoptosis.