|
|
|
Adenovirus-mediated tissue inhibitor of metalloproteinase-3
gene transfection inhibits rabbit intervertebral disc degeneration in vivo |
| Xudong YU MM,Zengwu SHAO MD,Liming XIONG MD,Weiwei XU MM,Hezhong WANG MM,Huifa XU MM, |
| Department of Orthopedics,
Union Hospital, Tongji Medical College, Huazhong University of Science
and Technology, Wuhan 430022, China; |
|
|
|
|
Abstract The aim of this study was to investigate the inhibitory effects of recombinant adenovirus vector carrying tissue inhibitor of metalloproteinase-3 (RAdTIMP-3) against degeneration of rabbit intervertebral disc. Thirty Japanese white rabbits of 4 months old were randomly divided into 5 groups. Mild or moderate rabbit lumbar disc degeneration model was constructed with the controllable axial loading device by imposing 98N pressure at the discs for 2 weeks. Various doses of virus were injected into the degenerated discs as follows: 20μL of normal saline in group 1; 20μL of RAd66 (an empty adenovirus vector, 1.0×1010OPU/mL) in group 2; and 20, 10, and 5μL of RAdTIMP-3 (1.0×1010OPU/mL) in groups 3, 4, and 5, respectively. Two weeks after the injection, the discs were collected for investigations, including assessment of degeneration degrees according to the Thompson’s grading system, reverse-transcription polymerase chain reaction (RT-PCR) assay for TIMP-3 gene, Safranin O-Fast green staining, and immunohistochemical staining for TIMP-3 and type II collagen. According to Thompson’s criteria, the degeneration of groups 3, 4, and 5, especially group 3, was alleviated as compared with groups 1 and 2. RT-PCR revealed that the expression of TIMP-3 in groups 3, 4, and 5, especially in group 3, was significantly enhanced as compared with group 1 (P<0.01). Both Safranin O-Fast green staining and type II collagen staining demonstrated better reserved integrity of disc matrix in groups 3, 4, and 5 than in groups 1 and 2. TIMP-3 staining exhibited an obvious increase of positive-staining rate in groups 3, 4, and 5 as compared with group 1. The positive-staining rate in group 3 (79.42%±1.35%) was about 3times that of group 1 (25.47%±5.46%, P<0.01). RAdTIMP-3 can effectively protect the matrix of rabbit intervertebral disc against overloading-induced degeneration in a dose-dependent manner, resulting in the alleviation of disc degeneration.
|
| Keywords
tissue inhibitor of metalloproteinase-3, intervertebral disc
rabbit
gene therapy
|
|
Issue Date: 05 December 2009
|
|
|
Arner E C, Pratta M A, Decicco C P, Xue C B, Newton R C, Trzaskos J M, Magolda R L, Tortorella M D. Aggrecanase: a target for the design of inhibitors ofcartilage degradation. Ann New York Sci, 1999, 878: 92―107
doi: 10.1111/j.1749-6632.1999.tb07676.x
|
|
Westling J, Fosang A J, Last K, Thompson V P, Tomkinson K N, Hebert T, McDonagh T, Collins-Racie LA, LaVallie E R, Morris E A, Sandy J D. ADAMTS4 cleavesat the aggrecanase site (Glu373-Ala374) and secondarily at the matrixmetalloproteinase site (Asn341-Phe342) in the aggrecan interglobulardomain. J Biol Chem, 2002, 277(18): 16059―16066
doi: 10.1074/jbc.M108607200
|
|
Little C B, Hughes C E, Curtis C L, Jones S A, Caterson B, Flannery C R. Cyclosporine A inhibition of aggrecanase-mediated proteoglycan catabolismin articular cartilage. Arthritis Rheum, 2002, 46(1): 124―129
doi: 10.1002/1529-0131(200201)46:1<124::AID-ART10121>3.0.CO;2-X
|
|
Sztrolovics R, Alini M, Roughley P J, Mort J S. Aggrecandegradation in human intervertebral disc and articular cartilage. Biochem J, 1997, 326(Pt 1): 235―241
|
|
Kashiwagi M, Tortorella M, Nagase H, Brew K. TIMP-3 is apotent inhibitor of aggrecanase 1 (ADAM-TS4) and aggrecanase 2 (ADAM-TS5). J Biol Chem, 2001, 276(16): 12501―12504
doi: 10.1074/jbc.C000848200
|
|
Malfait A M, Liu R Q, Ijiri K, Komiya S, Tortorella M D. Inhibition of ADAM-TS4 andADAM-TS5 prevents aggrecan degradation in osteoarthritic cartilage. J Biol Chem, 2002, 227(25): 222201―222208
|
|
Roberts S, Caterson B, Menage J, Evans E H, Jaffray D C, Eisenstein S M. Matrix metalloproteinases and aggrecanase: their rolein disorders of the human intervertebral disc. Spine, 2000, 25(23): 3005―3013
doi: 10.1097/00007632-200012010-00007
|
|
Roughley P J, Alini M, Antoniou J. The role of proteoglycans in aging, degeneration andrepair of the intervertebral disc. BiochemSoc Trans, 2002, 30(Pt 6): 869―874
|
|
Tsuji T, Chiba K, Imabayashi H, Fujita Y, Hosogane N, Okada Y, Toyama Y. Age-relatedchanges in expression of tissue inhibitor of metalloproteinases-3associated with transition from the notochordal nucleus pulposus tothe fibrocartilaginous nucleus pulposus in rabbit intervertebral disc. Spine, 2007, 32(8): 849―856
doi: 10.1097/01.brs.0000259804.39881.62
|
|
Patel K P, Sandy J D, Akeda K, Miyamoto K, Chujo T, An H S, Masuda K. Aggrecanasesand aggrecanase-generated fragments in the human intervertebral discat early and advanced stages of disc degeneration. Spine, 2007, 32(23): 2596―2603
doi: 10.1097/BRS.0b013e318158cb85
|
|
Pockert A J, Richardson S M, Le Maitre C L, Lyon M, Deakin J A, Buttle D J, Freemont A J, Hoyland J A. Modified expression of the ADAMTS enzymes and tissueinhibitor of metalloproteinases 3 during human intervertebral discdegeneration. Arthritis Rheum, 2009, 60(2): 482―491
doi: 10.1002/art.24291
|
|
Xiong X Q, Shao Z W, Pei H, Zhan Z R, Yu X D, Wang H Z. Construct and estimate of rabbit lumbar disc degeneration model withthe controllable axial loading. ZhongguoBingli Shengli Zazhi, 2008, 24(10): 2077―2080 (in Chinese)
|
|
Hu X J, Shao Z W. The in vivo effects of tumornecrosis factor-α on the degeneration of rabbit lumbar intervertebraldisc. Zhongguo Jizhu Jisui Zazhi, 2006, 16(7): 541―544 (in Chinese)
|
|
Thompson J P, Pearce R H, Schechter M T, Adams M E, Tsang I K, Bishop P B. Preliminary evaluation of a scheme for grading the gross morphologyof the human intervertebral disc. Spine, 1990, 15(5): 411―445
doi: 10.1097/00007632-199005000-00012
|
|
Xiong X, Yang S, Shao Z, Liu X, Zhan Z, Duan D. Up-regulationof niacinamide in intervertebral disc aggrecan in vitro. J Huazhong Univ Sci Technolog Med Sci, 2006, 26(1): 89―92
|
|
Xiong L M, Guo B, Shao Z W, Yang S H, Xie M, Wang H Z. Recombinant adenovirus carrying tissue inhibitor of metalloproteinase-3gene regulates the matrix of rabbit intervertebral disc in vivo. ZhongguoJiaoxing Waike Zazhi, 2009, 17(5): 356―360 (in Chinese)
|
|
Arner E C. Aggrecanase-mediated cartilage degradation. Curr Opin Pharmacol, 2002, 2(3): 322―329
doi: 10.1016/S1471-4892(02)00148-0
|
|
Okuda S, Myoui A, Ariga K, Nakase T, Yonenobu K, Yoshikawa H. Mechanisms of age-related decline in insulin-like growthfactor-I dependent proteoglycan synthesis in rat intervertebral disccells. Spine, 2001, 26(22): 2421―2426
doi: 10.1097/00007632-200111150-00005
|
|
Cs-Szabo G, Ragasa-San Juan D, Turumella V, Masuda K, Thonar E J, An H S. Changes in mRNA and proteinlevels of proteoglycans of the annulus fibrosus and nucleus pulposusduring intervertebral disc degeneration. Spine, 2002, 27(20): 2212―2209
doi: 10.1097/00007632-200210150-00006
|
|
Gendron C, Kashiwagi M, Hughes C, Caterson B, Nagase H. TIMP-3 inhibits aggrecanase-mediatedglycosaminoglycan release from cartilage explants stimulated by catabolicfactors. FEBS Lett, 2003, 555(3): 431―436
doi: 10.1016/S0014-5793(03)01295-X
|
|
Tsuji T, Chiba K, Imabayashi H, Fujita Y, Hosogane N, Okada Y, Toyama Y. Age-relatedchanges in expression of tissue inhibitor of metalloproteinases-3associated with transition from the notochordal nucleus pulposus tothe fibrocartilaginous nucleus pulposus in rabbit intervertebral disc. Spine, 2007, 32(8): 849―856
doi: 10.1097/01.brs.0000259804.39881.62
|
|
Le Maitre C L, Freemont A J, Hoyland J A. Localization of degradative enzymes and their inhibitorsin the degenerate human intervertebral disc. J Pathol, 2004, 204(1): 47―54
doi: 10.1002/path.1608
|
|
Johnstone B, Bayliss M T. The large proteoglycans ofthe human intervertebral disc. Changes in their biosynthesis and structurewith age, topography, and pathology. Spine, 1995, 20(6): 674―684
|
|
Moon S H, Gilbertson L G, Nishida K, Knaub M, Muzzonigro T, Robbins P D, Evans C H, Kang J D. Human intervertebral disc cells are geneticallymodifiable by adenovirus-mediated gene transfer: implications forthe clinical management of intervertebral disc disorders. Spine, 2000, 25(20): 2573―2579
doi: 10.1097/00007632-200010150-00006
|
|
Zhan Z, Shao Z, Xiong X, Yang S, Du J, Zheng Q, Wang H, Guo X, Liu Y. Ad/CMV- hTGF-beta1treats rabbit intervertebral discs degeneration in vivo. J Huazhong Univ SciTechnolog Med Sci, 2004, 24(6): 599―601, 624
|
|
MacLean J J, Roughley P J, Monsey R D, Alini M, Iatridis J C. Invivo intervertebral disc remodeling: kinetics of mRNA expressionin response to a single loading event. J Orthop Res, 2008, 26(5): 579―588
doi: 10.1002/jor.20560
|
|
Adams M A, Dolan P, Hutton W C, Porter R W. Diurnal changes in spinal mechanics and their clinical significance. J Bone Joint Surg, 1990, 72(2): 266―270
|
|
Gendron C, Kashiwagi M, Hughes C, Caterson B, Nagase H. TIMP-3 inhibits aggrecanase-mediatedglycosaminoglycan release from cartilage explants stimulated by catabolicfactors. FEBS Lett, 2003, 555(3): 431―436
doi: 10.1016/S0014-5793(03)01295-X
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
