| PrefaceIntervertebral disc degeneration ( IVDD) is known as that the intervertebral discs lost the normal structure and have a progressing fibrosis, displaying as disappearing of the gel like vertebral pulp , disappearing of the limes between thev-ertebral pulp and the fibrousring gradually , roughing and progressive fibrosisof the surrounding lamellae of the annulus fibrosus, finally appearing of the fissure and age pegment depositing. Pathological investigation of intervertebral disc degeneration has shown that the main change is the alteration of matrix, including structure , function, content and type of the proteoglycan, collagen, elastin . Intervertebral disc degeneration is one of main inducements of low back pain and intervertebral disc herniation. The data indicate the probability of low back pain is 60% -80% , low back pain is the third factor in the reason of adults losingla-bour force. But the investigation about the mechanism of intervertebral disc degeneration is not very clear.It has been noted that there are many factors synergistic effecting to result in the occuring of the interverlebral disc degeneration, including the lowernutri-tion of the vertebral disc, biomechanical factors, genetic factors, biochemistry-factors and etc. Effect of cytokines on IVDD are wildly mentioned, and the investigation is progressing currently. The purpose of our study is to detect the changes of the MMP-3,IL-1 contents in the degenerated vertebral discs and the fibrous rings , and then to illuminate the role of MMP-3,IL-1 during intervertebral disc degeneration and their interrelation.Materials and Methods1. Basic study1. 1 Animal grouping and sample gathering1.1.1 Animal selecting: we choose the Wistar rats at different age of 1 m, 3 m, 6 m, 12 m, 18 m and older, and 10 rats at the each age.1.1.2 Animal grouping: we choose the Wistar rats at different age of 1 m, 3ra,6m, 12 m, 18m and older, and 10 rats at the each age, so we have five trial groups.1.1.3 Sample gathering: after executing the rat by aeropleura we obtained the L4 - 5, L5 - 6 intervertebral discs as quickly as possible and fixed the discs in formalin and the 2.5% glutaral.1.2 Observation index and detection method1.2.1 Morphologic assessment the structure and appearance of the intervertebral disc cell in different age rats: HE and TEM1.2.2 The expression of MMP3 in the intervertebral disc cell at different age rats; SABC immunohistochemical method.1.3. Agent and instrumentMMP-3 monoclonal antibody,SABC immunohistochemical kit, DAB coloration fluid, polylysine(Poly-L-Lysine) ^deionized water^Leitzl512 slicer (Leitz, German) NBX60F/PM30 -3 H micrograph system(Olympus, Japan) ^Metamor-ph(NIC,U. S. A. ) /DP10/BX51(Olympus,Japan) color pathograph analytical system.Clinical Study2. 1. Selection and grouping of the sample;all the sample obtained from the secondary affiliated hospital of emu and FengTian hospitalNormal group: from the patient without PLCDAnnulus fibrosus incompletely disrupted group; the rupture of inner-layer of fibrous ring, the outer layer bulge by the Pressure from the vertebral pulpAnnulus fibrosus completely disrupted group: the rupture of full-layer of fibrous" ring, vertebral pulp protrude under the posterior longitudinal ligamentLiberation group: the rupture of full-layer fibrous ring, vertebral pulp in the vertebral canal2. 2. Observation index and detection methodthe expression of MMP3 and IL-1 in the intervertebral disc cell: Elisa method2. 3 Agent and instrumentElisa kit for MMP3^L-1 ^extration fluids electronic balance (Sartorius As) Ncentrifugal machine microipetResults1. Baisc study1. 1 Histology1.1.1 Oight microscope showing; Structural disorder of rat intervertebral disc increases gradually with age, viable count decreases gradually, collagen fibers appear hyperplasty, enlargement ,arrang disorder and break.1.1. 2Electron microscope: the number of the funicular cell and cartilage cell increases in the degenerated or necrosis intervertebral disc with the growth, there are many lipid droplet and glycogen deposition intracytoplasm, it can been find incomplete of cytomembrance, nuclear fragmentation, the formation of gran-ula-like mass in the exterior and interior of cell. The fibers are in hyperplasty in the exterior and interior of cell, and the fissure between those fiber can be find.1. 2Expression of MMP3 in the intervertebral disc of rat1.2. 1.Expression of MMP3 in the fibrous ring:The expression of the MMP3 in the fibrous ring of rat increases with the growth, the tendency has prominent statistical significance(P < 0. 01).1.2. 2 Expression of MMP3 in the vertebral pulp; The expression of the MMP3 in the vertebral pulp of rat increases with the growth, the tendency has promi-nent statistical significanee( P <0.01).2. Clinical studyExpression of MMP3NIL-1 in the intervertebral disc of human The expression of the MMP3 NIL-1 in intervertebral disc of PLCD patient is higher than the non-PLCd patient, the discrepancy has prominent statistical sig-... |
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