| Low back pain(LBP) is a common disease in orthopedical clinic which puzzles such a large amount of people and effect their lives and works.It is reported that over 80% of the population will suffer from LBP.As a report received from National Center for Health Statistics, LBP is the most common factor that causes older patient limit of activity.while the degeneration of disc is the most important factor which causes LBP . Many research results on the study of the pathophysiological mechanism of the intervertebral disc degeneration have been received. As a result , we know that the biochemical alterations in the disc have comparative strong relationship with the degeneration of intervertebral disc . But there are little research on the cartilaginous endplates of discs because of all kind of limits. In our study , we use an immunohistochemical method to research the expression of HSPs ( HSP27 &HSP72 ) in the discs in order to study the pathophysiological mechanism of HSPs in the degeneration process of human intervertebral discs.Materials and MethodsAll inpatients from Jan 2003 to Jan 2004, 15 patients of disc degeneration as Model group and 10 patients of spinal column trauma as Normal group were test. The patients in the Model group have the sign and symptom of that of the tipical herniation of disc , stenosis of spinal canal or spondylolisthesis. 7of 15 patients are the segment of L4-5 , the rest are L5-S1. Aging 48 to 68,with the average is 59.8. There are 10 male and 5 femal patients. They were diagnosed as 8 of spondylolisthesis , 5 of disc herniation , 2 of Low back pain of disc source. X-ray and MRI images are available to demonstrate the degeneration of the discs. 5 of all have the evidence of endplates degeneration. While the normal group denied of recurrent low back pain, X-ray,MRI,CT have no evidence of disc herniation. 5 of LI fracture, 3 of L2 fracture, 2 of L3 fracture, aging 30-46.with the average 37.5. There are 6 male and 4 femal patients. The specimens of endplates obtained during the operation of lumbar fusion. Other waste materials are eliminated including ligments , nucleus pulposus and annulus fibrosus. Thus the specimens are stored at -86The specimens are embedded in paraffin wax and slices are made from each specimen and transected at the center of each endplate. Then a immunohistochemical method is performed.immunostaining with antihuman HSP27 monoclonal antibody (clone 2B4 , Maixin_bio , fuzhou , china ; 1:100 dilution) and antihuman HSP72 polyclonal antibody(clone Maixin_bio , fuzhou , china ; 1:100 dilution) was performed. Negative controls were prepared by substituting the primary antibodies with buffer. Ten samples of spinal column trauma or fresh bodies were used as Normal controls.ResultsThe expression of HSP27 and HSP72 are positive which are obvious brown-yellow granules. It locates in the cytoplasm or nucleus, especially in the nucleus. All patients have evidence of HSP27 and HSP72 expression. In Normal group, the expression of HSP27 and HSP72 are 40% and 20%.while the Model group are 86.7% and 73.3% which has marked difference compared with the Normal group.As a result : HSPs can be found in the cytoplasm of the chondrocytes of cartilaginous endplates of normal people and enhance in that of the degenerative disc of those Model patients. The possible mechanisum of the expresson of HSPs are as below : the chondrocytes of cartilaginous endplates are in the precess of degeneration under long term stimulation of external or internal deleterious factors. The structure of proteins in the cells change and degernative and conglomerating proteins form which stimulates stress response of cell, activates heat shock gene and codic HSPs. The HSPs produced can accelerate the synthesize , collapse , assemble and locomotion of the protein in the cell. It helps the cells come back from stress so that redeuces injury of chondrocytes.ConclusionHSP27 and HSP72 were found in nucleus and cytoplasm of thecartilaginous endplates of normal people, especially in the nucleus. |
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