Investigation of VEGFA and HIF1-A gene expression levels in primary knee osteoarthritis patients

Abstract

Aim: Osteoarthritis (OA); is a disease characterized by biochemical and morphological changes caused by disruption of the balance between cartilage production and destruction. Angiogenesis and inflammation have been shown to be closely related processes to the development and progression of OA. Related research has shown that angiogenic factors such as vascular endothelial growth factor (VEGF) and hypoxia-induced factor (HIF) may play a role in the development of OA. The aim of this study was to determine the levels of VEGFA and HIF1A gene expression in peripheral mononuclear blood cells (PBMC) of OA patients and to determine the relationship between these genes and disease grades. Method: VEGFA and HIF1A mRNA expression in PBMCs of 97 patients with knee osteoarthritis and 41 healthy volunteers were investigated by quantitative real-time PCR (qRT-PCR). PBMCs were isolated from the peripheral blood samples from volunteers by density gradient centrifugation. Gene expression levels were determined by qRT-PCR using specific primer-probe sets after cDNA synthesis from the obtained cells. Results: There was a positive correlation (p <0.001) between VEGFA and HIF1A gene expressions between patient and control groups. In addition, VEGFA and HIF1A gene expression levels were not statistically different between the patient and control groups (p> 0.05) and between the grades of the disease (p> 0.05). Conclusion: In our study, it was shown that VEGFA and HIF1A gene expressions are correlated in OA PBMCs. Although the gene expression we studied was not sufficient as a biochemical marker in the diagnosis of knee osteoarthritis, it was concluded that it could serve as a potential therapeutic target with further studies.
Keywords: Osteoarthritis, VEGFA, HIF1A, PMKH, Gene Expression

Keywords

• Osteoarthritis,VEGFA,HIF1A,PMKH,Gene Expression

Primer diz osteoartrit hastalarında VEGFA ve HIF1-A gen ekspresyon seviyelerinin araştırılması

Abstract

Amaç: Osteoartrit (OA); kıkırdak yapımı ve yıkımı arasındaki dengenin bozulması ile ortaya çıkan, biyokimyasal ve morfolojik değişiklikler ile karakterize bir hastalıktır. Anjiyogenez ve inflamasyon süreçlerinin OA’in gelişmesi ve ilerlemesiyle ilişkili olduğu düşünülmektedir. Yapılan araştırmalar vasküler endotelial büyüme faktörü (VEGF) ve hipoksi ile indüklenen faktör (HIF) gibi anjiyogenik faktörlerin OA’nın gelişiminde rol oynayabileceğini göstermektedir. Bu çalışmada OA hastalarının peripheral mononüklear kan hücrelerinde (PMKH) VEGFA ve HIF1A gen ekspresyon seviyelerinin belirlemesi ve bu genlerin hastalığın evreleri ile olan ilişkisinin ortaya çıkarılması amaçlanmıştır. Yöntem: Diz osteoartriti tanısı almış 97 hasta ve 41 sağlıklı gönüllünün PMKH’lerinde VEGFA ve HIF1A mRNA ekspresyonları kantitatif real-time PCR (qRT-PCR) ile araştırıldı. Gönüllülerden alınan periferik kan örneklerinden PMKH'leri yoğunluk gradyanlı santrifüjleme ile izole edildi. Elde edilen hücrelerden cDNA sentezi sonrası spesifik primer-prob setleri kullanılarak qRT-PCR ile gen ekspresyon seviyeleri belirlendi. Bulgular: Hasta ve kontrol grupları arasında VEGFA ve HIF1A gen ekspresyonları arasında pozitif bir korelasyon (p<0.001) vardı. Bunun yanında, VEGFA ve HIF1A gen ekspresyon seviyelerinin, hasta ve kontrol grubu arasında (p>0.05) ve hastalığın evreleri arasında (p>0.05) istatistiksel olarak fark göstermediği belirlendi. Sonuç: Çalışmamızda OA PMKH’lerinde VEGFA ve HIF1A gen ekspresyonlarının birbiriyle ilişkili olduğu gösterilmiştir. Çalıştığımız gen ekspresyonları evrelerine göre diz osteoartritinin tanısında biyokimyasal marker olarak yeterli olamasa da, yapılacak ileri çalışmalarla potansiyel terapötik hedef olarak hizmet edebileceği kanısına varılmıştır.

Keywords

• Osteoartrit,VEGFA,HIF1A,PMKH,Gen ekspresyonu

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