Koroner Arter Hastalığı Olmayan Ve Statin İle Tedavi Edilen Bireylerde Ampk Yolağında Yeni Bir Aday Modülatör Olarak Microrna-625-5p
Amaç: AMP-aktive protein kinaz (AMPK) 3-hidroksi-3-metilglutaril koenzim A’yı fosforilleyen ve aynızamanda kolesterol biyosentezinin kontrolünde rol alan bir redüktazdır. Çalışmamızda, AMPK sinyal yolununaterosklerozdaki rolünü ve insan makrofaj hücrelerinde etkili olup olmadığını test etmeyi amaçladık. Buamaçla, koroner arter hastalığı (KAH) olmayan bireylerin plazma microRNA (miRNA) profillerinde statin varlığındaekspresyon değişimlerini göstermek ve AMPK yolunun yeni modülatörlerini tanımlamayı hedefledik.Yöntemler: Kolesterol düzeyine bağlı ekspresyon değişimini değerlendirmek için simvastatin ile uyarılanTHP-1 makrofajlarında, AMPK yoluna katılan seçilmiş genlerin ekspresyon düzeylerindeki değişikliklerbelirlendi. Agilent’in miRNA Mikroarray platformu, koroner anjiografi sonucunda koroner damarlarındakitıkanıklık oranı belirlenen statin kullanan ve kullanmayan bireylerin (≤30 koroner darlık) plazma miRNA profilinikarşılaştırmak için kullanıldı. MiRDB veritabanı, farklı ekprese olan miRNA’ların AMPK sinyal yolundarol oynayan hedef genlerini belirlemek için kullanıldı. Hedef genlerin ekspresyon değişimleri real-time PCRkullanılarak analiz edildi.Bulgular: Statin ile tedavi olan bireylerde, 5 miRNA’nın ekspresyon düzeylerinin anlamlı derecede azaldığısaptandı (miR-625-5p, miR-550a-3-5p, miR-550b-2-5p, miR-550a-5p, miR-let-7d-5p) (kat değişikliği >1.5, p <0.05). Buna ek olarak, simvastatinin THP-1 makrofajlarında AMPK α1, AMPK α2, AMPK β1, AMPK β2ve SREBF2 ekspresyonlarının artışına neden olduğu gözlendi. MİRDB kullanılarak, AMPK α1,’nin miR-625-5p için güçlü bir aday hedef olduğu gösterildi (Hedef Skor =95) .Sonuç: Elde ettiğimiz sonuçlar göre, statinin miR-625-5p ekspresyonunu azaltması AMPK α1’in indüklenmesiyolu ile kolesterol mekanizmalarının sentez yolunu aktive edebilir. Apoptoz ile ilişkili miR-625, ateroskleroz hastalarında statinlerin tedavi etkinliği için yeni bir belirteç olarak da kullanılabilir.
Microrna-625-5p as a Novel Candidate Modulator of Ampk Pathway in Statin-Treated Individuals Without Coronary Artery Disease
Aim: AMP-activated protein kinase (AMPK) is a signal molecule that phosphorylates and inactivates3-hydroxy-3-methylglutaryl coenzyme A reductase, the key enzyme controlling cholesterol biosynthesis.We aimed to test whether AMPK signaling pathway is effective in human macrophages which play crucialrole in atherosclerosis. In addition, to identify novel modulators of AMPK pathway while exploring the effectof statins on plasma microRNA (miRNA) profile of individuals without coronary artery disease (CAD).Medhod: We measured the expressions of selected genes involved in AMPK pathway in THP-1 macrophagestreated with simvastatin to assess dependence of their expression on cholesterol level. The Agilent’smiRNA Microarray analyses were performed to compare plasma miRNA profile of individuals (≤30%coronary stenosis) treated with and without statin. MiRDB target prediction tool was utilized to identifymiRNA target genes involved in AMPK pathway for differently expressed miRNAs. Expressions of targetgenes were analyzed by using real-time PCR.Results: Significantly decreased expression of 5 miRNAs (miR-625-5p, miR-550a-3-5p, miR-550b-2-5p,miR-550a-5p, miR-let-7d-5p) were observed in individuals receiving statins compared to non-statin group(fold change>1.5, p<0.05). In addition, simvastatin increased the expression of AMPK α1, AMPK α2, AMPKβ1, AMPK β2 and SREBF2 in THP-1 macrophages. Using miRDB target prediction tool, we identifed AMPKα1 is a strong candidate (target score=95) for miR-625-5p.Conclusion: Our results suggests that statin may downregulate the expression of miR-625-5p and induceAMPKα1 expression which in turn may activates cholesterol syntesis pathway. Apoptosis-associated miR-625 may be used as a novel biomarker for effectiveness of statins therapy in patients with atherosclerosis.
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