Research Paper Volume 13, Issue 3 pp 4299—4316

Overexpression of long noncoding RNA ANRIL inhibits phenotypic switching of vascular smooth muscle cells to prevent atherosclerotic plaque development in vivo

Overexpression of lncRNA-ANRIL abolishes PDGF-induced phenotypic switching and reverses AMPK activity in cultured VSMCs. Cultured VSMCs were infected with adenovirus expressing lncRNA-ANRIL for 24 hours and then treated with PDGF (5 ng/ml) for 48 hours. (A) The expression of lncRNA-ANRIL was determined by real-time PCR. N is 5 in each group. *PB–G) The mRNA levels of the markers of VSMC phenotypic switching, including α-SMA in B, calponin in C, smoothelin in D, osteopontin in E, collagen I in F, and collagen III in G were measured by real-time PCR. (H) Total cell lysates were subjected to perform western blot analysis of phosphorylated AMPK (pAMPK) and total AMPK protein levels. (I) AMPK activity was assayed by P32-ATP method. N is 5 in each group. *P#P

Figure 2. Overexpression of lncRNA-ANRIL abolishes PDGF-induced phenotypic switching and reverses AMPK activity in cultured VSMCs. Cultured VSMCs were infected with adenovirus expressing lncRNA-ANRIL for 24 hours and then treated with PDGF (5 ng/ml) for 48 hours. (A) The expression of lncRNA-ANRIL was determined by real-time PCR. N is 5 in each group. *P<0.05 vs. adenovirus vector. (BG) The mRNA levels of the markers of VSMC phenotypic switching, including α-SMA in B, calponin in C, smoothelin in D, osteopontin in E, collagen I in F, and collagen III in G were measured by real-time PCR. (H) Total cell lysates were subjected to perform western blot analysis of phosphorylated AMPK (pAMPK) and total AMPK protein levels. (I) AMPK activity was assayed by P32-ATP method. N is 5 in each group. *P<0.05 vs. adenovirus alone. #P<0.05 vs. adenovirus + PDGF.