개의 대퇴골두인대 절단 모델에서 프로테오믹스로 관찰한 관절액의 변화

Proteomic-determined Alteration of Synovial Fluid on Induced Model of Transected Ligament of Head of Femur

  • 김세훈 (전북대학교 수의과대학, 생체안전성연구소) ;
  • 신기욱 (전북대학교 수의과대학, 생체안전성연구소) ;
  • 지중룡 (백광 C & S) ;
  • 심관섭 (전북대학교 농업생명과학대학) ;
  • 김남수 (전북대학교 수의과대학, 생체안전성연구소)
  • Kim, Se-Hoon (College of Veterinary Medicine and Biosafety Research Institute) ;
  • Shin, Ki-Uk (College of Veterinary Medicine and Biosafety Research Institute) ;
  • Ji, Joong-Ryong (Paek kwang C&S Co. LTD.) ;
  • Shim, Kwan-Seob (College of Agriculture and Life Science, Chonbuk National University) ;
  • Kim, Nam-Soo (College of Veterinary Medicine and Biosafety Research Institute)
  • 심사 : 2010.12.13
  • 발행 : 2010.12.31

초록

관절염은 증상 발현 이후 치료 효과가 매우 더디게 나타나기 때문에 조기 발견과 그에 따른 적절한 처치가 무엇보다도 중요하다. 따라서 많은 연구자들은 관절염 질환모델을 통한 조기 진단 인자의 발견을 위해 그 동안 지속적으로 연구해 오고 있는 실정이다. 개에서 고관절 탈구와 이형성은 정형외과에서 흔한 질병으로 고관절의 관절염과 관련이 있는 것으로 알려져 있다. 관절염 유발을 위해서는 흔히 대퇴골두인대의 절단이 필요한데 이는 대퇴골두인대의 절단을 통해 관절의 움직임을 극대화 시키기 위한 방법이다. 지금까지 유발 관절염에 대한 관절액의 분석을 위해서 프로테오믹스를 이용한 연구는 매우 드물다. 실험동물은 열 마리의 건강한 수컷 비글견을 사용하였으며, 나이는 2-3세 (평균 $2.57{\pm}0.20$ 살), 몸무게는 10 - 16kg (평균 $11.90{\pm}1.68 kg$) 이었다. 실험 기간 동안 방사선검사, 일반혈액검사, 혈청화학검사상 특이사항이 없었다. 실험동물은 술 후 평균 일주일에 보행을 하였다. Proteomic 검사 상, 관절액의 단백질 변화가 보여졌으며 단백질의 변화의 유의성검증을 위해 ANOVA (p < 0.05)테스트를 실시하였다. 실험 결과로는 프로테오믹스 검사에서는2개의 단백질 점(Vitamin D-binding protein precursor, ANOVA (p < 0.004); Kinogen-1, ANOVA (p < 0.039))이 유의성 있게 증가되는 모습이 확인하였다(ANOVA (p < 0.05)). 이 두 단백질은 류마티스 관절염, 혹은 퇴행성 관절염에서도 증가하는 경향이 있는 것으로 알려져 있다. 이 실험에서 관절염 유발 방법으로 이용한 대퇴골두인대의 절단은 관절염 유발에 적절한 모델이었으며 또한 고관절에서 관절액 단백질의 변화를 유도할 수 있는 적합한 방법이라고 생각된다. 따라서 관절염 유발 모델에서 프로테오믹스를 이용한 관절액 변화의 관찰은 실제 임상에 있어서 관절염 조기 진단방법으로 매우 유용성이 있을 것으로 생각된다.

Many animal models of osteoarthritis (OA) have been developed, aimed at understanding the long-term progression of OA and difficulty of identifying patients in the initial stage of the disease. In canines, coxofemoral luxation and hip dysplasia are common orthopedic ailments related to OA in the hip joint. Transecting the ligament of the head of the femur (LHF) aids in diagnosis of coxofemoral joint OA. Presently, mobility of this joint was increased by transected LHF in 10 mature, 2-3-year-old (average $2.57{\pm}0.20$ years), healthy male beagles. The animals were normally gaited 1-week post-operatively. During the experimental period, examinations including X-ray, complete blood count and serum chemistry were unremarkable. Proteomic examination revealed protein alterations in synovial fluid, with significant increases in Vitamin D-binding protein precursor (ANOVA, p < 0.004) and Kinogen-1 (ANOVA, p < 0.039). Both proteins correlated with arthritis.

키워드

참고문헌

  1. Breslow J, Ross D, McPherson J, Williams H, Kurnit D, Nussbaum A, Karathanasis S, Zannis V. Isolation and characterization of cDNA clones for human apolipoprotein AI. Proc Natl Acad Sci USA 1982; 79: 6861-6865. https://doi.org/10.1073/pnas.79.22.6861
  2. Burger D, Dayer J. High-density lipoprotein-associated apolipoprotein AI: the missing link between infection and chronic inflammation? Autoimmunity Reviews 2002; 1: 111-117. https://doi.org/10.1016/S1568-9972(01)00018-0
  3. Cassim B, Shaw O, Mazur M, Misso N, Naran A, Langlands D, Thompson P, Bhoola K. Kallikreins, kininogens and kinin receptors on circulating and synovial fluid neutrophils: role in kinin generation in rheumatoid arthritis. Rheumatology 2009; 48: 490-496.
  4. De Ceuninck F, Berenbaum F. Proteomics: addressing the challenges of osteoarthritis. Drug Discov Today 2009; 14: 661-667. https://doi.org/10.1016/j.drudis.2009.05.006
  5. Girling S, Bell S, Whitelock R, Rayward R, Thomson D, Carter S, Vaughan TA, Innes J. Use of biochemical markers of osteoarthritis to investigate the potential disease-modifying effect of tibial plateau levelling osteotomy. J Small Anim Pract 2006; 47: 708-714. https://doi.org/10.1111/j.1748-5827.2006.00150.x
  6. Gobezie R, Kho A, Krastins B, sarracino D, Thornhill T, Chase M, Millett P, Lee D. High abundance synovial fluid proteome: distinct profiles in health and osteoarthritis. Arthritis Res Ther 2007; 9: 36.
  7. Hayes M, Denton J, Freemont A, Mawer E. Synthesis of the active metabolite of vitamin D, 1, 25 (OH) 2D3, by synovial fluid macrophages in arthritic diseases. Ann Rheum Dis 1989; 48: 723-729. https://doi.org/10.1136/ard.48.9.723
  8. Ishioka N, Takahashi N, Putnam F. Amino acid sequence of human plasma alpha 1B-glycoprotein: homology to the immu-noglobulin supergene family. Proc Natl Acad Sci USA 1986; 83: 2363-2367. https://doi.org/10.1073/pnas.83.8.2363
  9. Jethwaney D, Lepore T, Hassan S, Mello K, Rangarajan R, Jahnen-Dechent W, Wirth D, Sultan A. Fetuin-A, a hepatocytespecific protein that binds Plasmodium berghei thrombospondinrelated adhesive protein: a potential role in infectivity. Infect Immun 2005; 73: 5883-5891. https://doi.org/10.1128/IAI.73.9.5883-5891.2005
  10. Kong S, Stabler T, Criscione L, Elliott A, Jordan J, Kraus V. Diurnal variation of serum and urine biomarkers in patients with radiographic knee osteoarthritis. Arthritis Rheum 2006; 54: 2496-2504. https://doi.org/10.1002/art.21977
  11. Langlois M, Delanghe J. Biological and clinical significance of haptoglobin polymorphism in humans. Clin Chem 1996; 42: 1589-1600.
  12. Lascelles B, King S, Roe S, Marcellin-Little D, Jones S. Expression and activity of COX-1 and 2 and 5-LOX in joint tissues from dogs with naturally occurring coxofemoral joint osteoarthritis. J Orthop Res 2009; 27: 1204-1208. https://doi.org/10.1002/jor.20864
  13. Mande J, Mbithi P, Mbugua S, Buoro I, Gathumb Pi. Volume of the ligamentum capitis femoris in osteoarthritic hip joints of adult dogs. J S Afr Vet Assoc 2003; 74: 11-13.
  14. Nganvongpanit K, Itthiarbha A, Ong-Chai S, Kongtawelert P. Evaluation of serum chondroitin sulfate and hyaluronan: biomarkers for osteoarthritis in canine hip dysplasia. J Vet Sci 2008; 9: 317-325. https://doi.org/10.4142/jvs.2008.9.3.317
  15. Ohta N, Sato M, Ushida K, Kokubo M, Baba T, Taniguchi K, Urai M, Kihira K, Mochida J. Jellyfish mucin may have potential disease-modifying effects on osteoarthritis. BMC Biotechnol 2009; 9: 98. https://doi.org/10.1186/1472-6750-9-98
  16. Ozaydin, Kilic E, Baran V, Demirkan, Kamiloglu A, Vural S. Reduction and stabilization of hip luxation by the transposition of the ligamentum sacrotuberale in dogs: an in vivo study. Vet Surg 2003; 32: 46-51. https://doi.org/10.1053/jvet.2003.50009
  17. Quaye I. Haptoglobin, inflammation and disease. Trans R Soc Trop Med Hyg 2008; 102: 735-742. https://doi.org/10.1016/j.trstmh.2008.04.010
  18. Seol J, Lee H, Lee Y, Lee Y, Kang H, Kim I, Kim N, Park S. Hypoxic resistance to articular chondrocyte apoptosis-a possible mechanism of maintaining homeostasis of normal articular cartilage. FEBS J 2009; 276: 7375-7385. https://doi.org/10.1111/j.1742-4658.2009.07451.x
  19. Slatter D. Hip Dysplasia: Diagnosis and Nonsurgical Treatment. In: Textbook of small animal surgery, 3rd ed. Philadelphia: Saunders. 2002: 2009-2018.
  20. Vos P, Interma F, van El B, DeGroot J, Bijlsma J, Lafeber F, Mastbergen S. Does loading influence the severity of cartilage degeneration in the canine Groove-model of OA? J Orthop Res 2009: 27: 1332-1338. https://doi.org/10.1002/jor.20897
  21. Wakitani S, Okabe T, Kawaguchi A, Nawata M, Hashimoto Y. Highly sensitive ELISA for determining serum keratan sulphate levels in the diagnosis of OA. Rheumatology 2010: 49: 57-62. https://doi.org/10.1093/rheumatology/kep351
  22. Walker J. Growth characteristics of the fetal ligament of the head of femur: significance in congenital hip disease. Yale J Biol Med 1980; 53: 307-316.
  23. White P, Cooke N. The multifunctional properties and characteristics of vitamin D-binding protein. Trends Endocrinol Metab 2000; 11: 320-327. https://doi.org/10.1016/S1043-2760(00)00317-9
  24. Williams R, Henderson L, Naidoo Y, Cassim B, Elson C, Bhoola K. Immunocytochemical analysis of tissue kallikrein and the kinin moiety in rheumatoid synovial fluid neutrophils. Rheumatology 1997; 36: 420-425. https://doi.org/10.1093/rheumatology/36.4.420
  25. Wilson R, Whitelock J, Bateman J. Proteomics makes progress in cartilage and arthritis research. Matrix Biol 2009; 28: 121-128. https://doi.org/10.1016/j.matbio.2009.01.004
  26. Yi J, Chang J, Han W, Lee J, Ko E, Kim D, Bae J, Yu J, Lee C, Yu M. Autoantibody to tumor antigen, alpha 2-HS glycoprotein: a novel biomarker of breast cancer screening and diagnosis. Cancer Epidemiol Biomarkers Prev 2009; 18: 1357-1364. https://doi.org/10.1158/1055-9965.EPI-08-0696
  27. Zannis V, Kurnit D, Breslow J. Hepatic apo-AI and apo-E and intestinal apo-AI are synthesized in precursor isoprotein forms by organ cultures of human fetal tissues. J Biol Chem 1982; 257: 536-544.