mersin univ saglık bilim derg

Mersin Üniversitesi Sağlık Bilimleri Dergisi

1308-0830

Mersin University

10.26559/mersinsbd.628048

Health Care Administration

Sağlık Kurumları Yönetimi

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

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

https://orcid.org/0000-0003-0999-2774

Soyocak

Ahu

İSTANBUL AYDIN ÜNİVERSİTESİ, TIP FAKÜLTESİ

https://orcid.org/0000-0002-8488-6405

Turgut Coşan

Didem

ESKİŞEHİR OSMANGAZİ ÜNİVERSİTESİ, TIP FAKÜLTESİ

https://orcid.org/0000-0002-8919-968X

Özgen

Merih

ESKİŞEHİR OSMANGAZİ ÜNİVERSİTESİ, TIP FAKÜLTESİ

https://orcid.org/0000-0003-2433-9925

Kurt

Hülyam

ESKİŞEHİR OSMANGAZİ ÜNİVERSİTESİ, TIP FAKÜLTESİ

https://orcid.org/0000-0002-9339-4031

Şahin Mutlu

Fezan

ESKİŞEHİR OSMANGAZİ ÜNİVERSİTESİ, TIP FAKÜLTESİ

04 30 2020

13 1 97 106 10 01 2019 02 13 2020

2008

Mersin Üniversitesi Sağlık Bilimleri Dergisi

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

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.

Osteoartrit VEGFA HIF1A PMKH Gen ekspresyonu

Osteoarthritis VEGFA HIF1A PMKH Gene Expression

1. Sharma L, Kapoor D, Issa S. Epidemiology of osteoarthritis: an update. Current opinion in rheumatology. 2006;18(2):147–156.

2. Pelletier J, Martel‐Pelletier J, Abramson SB. Osteoarthritis, an inflammatory disease: potential implication for the selection of new therapeutic targets. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology. 2001;44(6):1237–1247.

3. Saxne T, Lindell M, Månsson B, Petersson IF, Heinegård D. Inflammation is a feature of the disease process in early knee joint osteoarthritis. Rheumatology (Oxford, England). 2003;42(7):903.

4. Van der Kraan PM, van den Berg WB. Anabolic and destructive mediators in osteoarthritis. Current Opinion in Clinical Nutrition & Metabolic Care. 2000;3(3):205–211.

5. Dennison E. Osteoarthritis epidemiology and classification. Rheumatology. 2003:1781–1792.

6. Tuncer T. Gilgil E. Osteoartrit epidemiyolojisi ve risk faktörleri, Tanıdan tedaviye osteoartrit. Nobel Tıp Kitabevleri, İstanbul. 2007:9–20.

7. Bodur H. Dünyada ve Türkiye’de Osteoartrite Güncel Bakiş; Epidemiyoloji ve Sosyoekonomik Boyut. Türk Geriatri Dergisi. 2011;14(Özel Sayı 1):7–14.

8. Patogenezi DCO. Kelley Romatoloji. Ankara Güneş Kitapevi. 2006:1493–1513.

9. Bonnet CS, Walsh DA. Osteoarthritis, angiogenesis and inflammation. Rheumatology. 2005;44(1):7–16.

10. Schutze N. Angiogenesis in osteoarthritis. Current Rheumatology Reviews. 2008;4(3):206–209.

11. Lingen MW. Role of leukocytes and endothelial cells in the development of angiogenesis in inflammation and wound healing. Archives of pathology & laboratory medicine. 2001;125(1):67–71.

12. Cohen T, Nahari D, Cerem LW, Neufeld G, Levi B-Z. Interleukin 6 induces the expression of vascular endothelial growth factor. Journal of Biological Chemistry. 1996;271(2):736–741.

13. Ben-Av P, Crofford LJ, Wilder RL, Hla T. Induction of vascular endothelial growth factor expression in synovial fibroblasts by prostaglandin E and interleukin-1: a potential mechanism for inflammatory angiogenesis. FEBS letters. 1995;372(1):83–87.

14. Hoeben ANN, Landuyt B, Highley MS, Wildiers H, Van Oosterom AT, De Bruijn EA. Vascular endothelial growth factor and angiogenesis. Pharmacological reviews. 2004;56(4):549–580.

15. Yazır Y, Gonca S, Filiz S, Dalçık H. Endotel hücreleri için önemli bir protein ailesi; vasküler endotel büyüme faktörü (VEGF), Ailenin üyeleri, yapısı ve sentezi. Cumhuriyet Üniversitesi Tıp Fakültesi Dergisi. 2004;26:181–184.

16. Neve A, Cantatore FP, Corrado A, Gaudio A, Ruggieri S, Ribatti D. In vitro and in vivo angiogenic activity of osteoarthritic and osteoporotic osteoblasts is modulated by VEGF and vitamin D3 treatment. Regulatory peptides. 2013;184:81–84.

17. Çebi H, Aksahin E, Yuksel HY, Çelebi L, Aktekin CN, Hapa O, Muratli HH, Bicimoglu A. Plasma vascular endothelial growth factor levels are similar in subjects with and without osteoporosis. Eklem Hastalik Cerrahisi. 2010;21(2):91–97.

18. Saetan N, Honsawek S, Tanavalee A, Yuktanandana P, Meknavin S, Ngarmukos S, Tanpowpong T, Parkpian V. Relationship of plasma and synovial fluid vascular endothelial growth factor with radiographic severity in primary knee osteoarthritis. International orthopaedics. 2014;38(5):1099–1104.

19. Ballara S, Taylor PC, Reusch P, Marmé D, Feldmann M, Maini RN, Paleolog EM. Raised serum vascular endothelial growth factor levels are associated with destructive change in inflammatory arthritis. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology. 2001;44(9):2055–2064.

20. Qing L, Lei P, Liu H, Xie J, Wang L, Wen T, Hu Y. Expression of hypoxia-inducible factor-1α in synovial fluid and articular cartilage is associated with disease severity in knee osteoarthritis. Experimental and therapeutic medicine. 2017;13(1):63–68.

21. Şahin Calapoğlu N. HIF-1: İki Ucu Keskin Bıçak. Smyrna Tıp Dergisi. 2016;2:54–58.

22. Wenger RH. Cellular adaptation to hypoxia: O2-sensing protein hydroxylases, hypoxia-inducible transcription factors, and O2-regulated gene expression. The FASEB journal. 2002;16(10):1151–1162.

23. Hamilton JL, Nagao M, Levine BR, Chen D, Olsen BR, Im H. Targeting VEGF and its receptors for the treatment of osteoarthritis and associated pain. Journal of Bone and Mineral Research. 2016;31(5):911–924.

24. Jian-Lin Z, Hong-Song F, Hao P, Shuang D, Shen C, Jian-Ping L, Bo Q, Jin-Qing W, Feng L. The relationship between HIF-2α and VEGF with radiographic severity in the primary osteoarthritic knee. Yonsei medical journal. 2016;57(3):735–740.

25. Altman R, Asch E, Bloch D, Bole G, Borenstein D, Brandt K, Christy W, Cooke TD, Greenwald R, Hochberg M. Development of criteria for the classification and reporting of osteoarthritis: classification of osteoarthritis of the knee. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology. 1986;29(8):1039–1049.

26. Kellgren JH, Lawrence JS. Radiological assessment of osteo-arthrosis. Annals of the rheumatic diseases. 1957;16(4):494.

27. Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C T method. Nature protocols. 2008;3(6):1101.

28. Yuan Q, Sun L, Li J-J, An C-H. Elevated VEGF levels contribute to the pathogenesis of osteoarthritis. BMC musculoskeletal disorders. 2014;15(1):437.

29. Abdel-Magied RA, AbuOmar HAS, Monir A, Higazi AA, Amal A. Synovial fluid level of vascular endothelial growth factor (VEGF) can predict functional status and radiological severity in patients with knee osteoarthritis (OA). Int. J. Clin. Rheumatol. 2019;14(1).

30. Zhu J, Tang Y, Wu Q, Ji Y, Feng Z, Kang F. HIF‐1α facilitates osteocyte‐mediated osteoclastogenesis by activating JAK2/STAT3 pathway in vitro. Journal of cellular physiology. 2019;234(11):1–11.

31. Fernández-Torres J, Martínez-Nava GA, Zamudio-Cuevas Y, Martínez-Flores K, Gutiérrez-Ruíz MC, Gómez-Quiroz LE, Garrido-Rodríguez D, Muñoz-Valle JF, Oregón-Romero E, Lozada C. Impact of the gene-gene interactions related to the HIF-1α signaling pathway with the knee osteoarthritis development. Clinical Rheumatology. 2019:1–11.

32. Murata M, Yudoh K, Nakamura H, Kato T, Inoue K, Chiba J, Nishioka K, Masuko‐Hongo K. Distinct signaling pathways are involved in hypoxia‐and IL‐1‐induced VEGF expression in human articular chondrocytes. Journal of orthopaedic research. 2006;24(7):1544–1554.

33. Chen Y, Zhao B, Zhu Y, Zhao H, Ma C. HIF-1-VEGF-Notch mediates angiogenesis in temporomandibular joint osteoarthritis. American Journal of Translational Research. 2019;11(5):2969.