Koroner Arter Hastalığı Olmayan Ve Statin İle Tedavi Edilen Bireylerde Ampk Yolağında Yeni Bir Aday Modülatör Olarak Microrna-625-5p

Yıl 2016, Cilt: 6 Sayı: 12, 11 - 10, 01.12.2016

Neslihan Çoban , Dilek Pirim Aycan Fahri Erkan Filiz Güçlü-geyik Berkay Ekici Nihan Erginel-ünaltuna

Öz

Amaç: AMP-aktive
protein kinaz (AMPK) 3-hidroksi-3-metilglutaril koenzim A’yı fosforilleyen ve
aynı

zamanda kolesterol biyosentezinin
kontrolünde rol alan bir redüktazdır. Çalışmamızda, AMPK sinyal yolunun

aterosklerozdaki rolünü ve insan
makrofaj hücrelerinde etkili olup olmadığını test etmeyi amaçladık. Bu

amaçla, koroner arter hastalığı
(KAH) olmayan bireylerin plazma microRNA (miRNA) profillerinde statin varlığında

ekspresyon değişimlerini göstermek
ve AMPK yolunun yeni modülatörlerini tanımlamayı hedefledik.

Yöntemler: Kolesterol
düzeyine bağlı ekspresyon değişimini değerlendirmek için simvastatin ile uyarılan

THP-1 makrofajlarında, AMPK yoluna
katılan seçilmiş genlerin ekspresyon düzeylerindeki değişiklikler

belirlendi. Agilent’in miRNA
Mikroarray platformu, koroner anjiografi sonucunda koroner damarlarındaki

tıkanıklık oranı belirlenen statin
kullanan ve kullanmayan bireylerin (≤30 koroner darlık) plazma miRNA profilini

karşılaştırmak için kullanıldı.
MiRDB veritabanı, farklı ekprese olan miRNA’ların AMPK sinyal yolunda

rol oynayan hedef genlerini
belirlemek için kullanıldı. Hedef genlerin ekspresyon değişimleri real-time PCR

kullanılarak analiz edildi.

Bulgular: Statin
ile tedavi olan bireylerde, 5 miRNA’nın ekspresyon düzeylerinin anlamlı
derecede azaldığı

saptandı (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 β2

ve 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
indüklenmesi

yolu ile kolesterol mekanizmalarının
sentez yolunu aktive edebilir. Apoptoz ile ilişkili miR-625,

ateroskleroz hastalarında
statinlerin tedavi etkinliği için yeni bir belirteç olarak da kullanılabilir.

Anahtar Kelimeler

AMPK, miR-625-5p, ateroskleroz, statin, kolesterol

Microrna-625-5p as a Novel Candidate Modulator of Ampk Pathway in Statin-Treated Individuals Without Coronary Artery Disease

Öz

Aim: AMP-activated protein kinase (AMPK) is a signal
molecule that phosphorylates and inactivates

3-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 crucial

role in atherosclerosis. In
addition, to identify novel modulators of AMPK pathway while exploring the
effect

of 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 macrophages

treated with simvastatin to
assess dependence of their expression on cholesterol level. The Agilent’s

miRNA 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 identify

miRNA target genes involved in
AMPK pathway for differently expressed miRNAs. Expressions of target

genes 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, AMPK

β1, AMPK β2 and SREBF2 in THP-1
macrophages. Using miRDB target prediction tool, we identifed AMPK

α1 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 induce

AMPKα1 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.

Anahtar Kelimeler

AMPK, miR-625-5p, atherosclerosis, statin, cholesterol

Kaynakça

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