Screen And Verification Of The Potential Mirnas Implicating In Aortic Calcification Progression Of OPG^-/- Mouse

Objectives : To screen the mi RNA profile during the process of vascular calcification in OPG-/- mouse, and to verify the potential mi RNAs witch could regulate vascular calcification.Methods : The mi RNA profile during the process of vascular calcification was screened by using mi RCURY? LNA Array(v.18.0)(Exiqon) with aortas of the WT mice and OPG-/- mice at 12 weeks of age and with the smooth muscle cell calcification model. QRT-PCR and in situ hybridization were utilized to verify the mi RNAs showing the common change trends in both in vivo and in vitro models.Results : Sixteen potential mi RNAs including mi R-125b-5p,mi R-30a-5p, mi R-32-5p, mi R-133a-3p and mi R-29a-5 were identified to be involved in vascular calcification progression from the in vivo and in vitro models.Verification of the key mi RNAs:1. Quantitative PCR analysis showed that seven of eight mi RNAs were validated to be consistent with the trends in the microarray analysis.The expression of mi R-125 b,mi R-30 a and mi R-32 were confirmed to be elevated, and mi R-133 a,mi R-29 a,mi R-210 and mi R-320 were proved to be decreased in the aortas of OPG-/- mice with vascular calcification(p<0.05).2. In situ hybridization confirmed that mi R-32 located mainly in the media of aorta and showed a slightly and markedly increased expression in aortas of OPG-/- mice at 4 and 12 weeks of age respectively, compared to those in WT mice.Conclusion : The 7 mi RNAs including mi R-125b-5p, mi R-32-5p,mi R-30a-5p, mi R-29a-5p, mi R-133a-3p, mi R-210-3P and mi R-320-3p were identified to be implicated in the progression of vascular calcification, and those mi RNAs might be served as candidates for biological marker and modulator of vascular calcification.