| Background: Lumbar degenerative diseases is lumbar spine of the pathological process of natural aging, degradation, spine surgery is one of the more common diseases.(lumbar disease, lumbar intervertebral disc protrusion lumbar disc herniation, LDH) is the most common clinical disease. LDH due to differences in age, gender, sick time and place, its clinical manifestations and clinical symptoms are also different, according to its clinical can be classified into category of lumbago, poliomyelitis and other traditional Chinese medicine. Development in recent years, the disease is on the rise, and the trend has gradually expand from the older adults to young adults trend of lumbar disc degeneration is the pathological basis lumbar degenerative disease, lumbar disc degenerative disease characterized by the degradation of extracellular matrix(extracellularmatrix, ECM), intervertebral disc cells reduce; Lumbar mechanical property change is the cause of the extracellular matrix degradation, thus inducing hernia of lumbar disc herniation, and further cause the corresponding signs and symptoms. So far, the medicine understanding of the cause of the intervertebral disc degeneration is not fully clear, can determine its factors include a variety of, such as hereditary, abnormal cytokines, patients’ age characteristics change, matrix metalloproteinases(matrixmetall oproteinases, MMPs) activity increased abnormal,biomechanical change and other factors, are involved in the process of intervertebral disc degeneration. Is thought to play a key role in the process of intervertebral disc degeneration is the exceptional increasing on the activity of MMPs. It can reduce the function of extracellular matrix membrane composition. Since found the first kind of MMPs, to unknown at present, a total of 28 separated in MMPs family member, found the MMPs family members according to the sequence was found by naming, respectively named MMP-1 ~ MMP-28. Normal human intervertebral disc tissue, contains a small amount of MMPs and MMPs inhibitor(TIMPs), there is a certain proportion relationship between MMPs and TIMPs expression, together they maintain the balance of the synthesis and degradation of the ECM. In the degeneration of intervertebral disc tissue, MMP-3 and MMP-9 affected by many factors such as cytokines and their regulating control, active expression significantly higher, so the two members of the family of MMPs is considered to be an important enzyme in organization of lumbar disc degeneration, and degeneration of intervertebral disc tissue has a close relationship. In check diagnosis of intervertebral disc degeneration, MRI(magnetic resonance imaging, MRI) is one of the main methods of imaging examination is the examination diagnosis method has a very wide range of application in clinic. In is the classification standard of intervertebral disc degeneration, the spine surgeon Pfirrmann grading standard is the common recognition of intervertebral disc degeneration classification standard. At present, the academia has not appeared on MMPs in different Pfirrmann grading expression of intervertebral disc degeneration in the related research.Objective: Based on the above background, the purpose of this study lies in: 1) through the detection of MMP-3 and MMP-9 in normal or abnormal expression of disc nucleus pulposus tissue, observe its expression whether there is a difference, 2) through the detection of MMP-3 and MMP-9 in normal or abnormal expression of disc nucleus pulposus tissue situation, analyze the existence of correlation between them; 3) in accordance with the approved of intervertebral disc degeneration Pfirrmann gradingstandard of classification, observe Pfirrmann grading standard under different classification of intervertebral disc degeneration of MMPS-3, presence of differences in the expression of MMP-9, if there are differences, analysis of the specific condition and reason for the difference.Methods: Select the first affiliated hospital of zhengzhou university bone five families from June 2013 to December 2013 patients admitted during the period of lumbar intervertebral disc protrusion, 46 cases take the specimen of nucleus pulposus degeneration group. Specimens of 46 cases of nucleus pulposus, from the 21 cases of male patients, from 25 cases of female patients, patients aged between 30 ~ 62, the average age was(42.6±2.2), including 2 cases of L2 / L3 and 3 cases of L3 / L4, 18 cases of L4 / L5, 23 cases of L5 / S1. Degeneration group all patients according to the results of MRI, to lumbar disc type further grouping, 23 cases of outstanding type as outstanding group, 16 cases of emergence as out groups, 7 cases of free model for free. Another choice during the same period because of the lumbar spine fractures in our undergraduate course room treatment of 12 cases of normal lumbar nucleus pulposus tissue samples for the control group. 12 cases of lumbar nucleus pulposus tissue samples from the male 7 cases, 5 cases from women, aged between 17 ~ 36 years old, the average age was(26.8±1.8), among them 6 cases of L1 / L2, L2 / L3 in 4, 2 cases of L3 / L4. Degeneration of nucleus pulposus tissue group and control group two groups of patients are in operation by the same set of surgery doctors take out, take out after wash with physiological saline in formaldehyde solution concentration(10%) in the soaking, immersion time of 24 h, department of pathology, the first after soaking send, fixation, dehydration, paraffin slice, the conventional processing, the nucleus pulposus tissue samples made into slices, each nucleus pulposus tissue samples made six slices, each slice thickness for 5 μm), slice in 0 ~ 4 ℃ refrigerator save for later use. Prepare good section line HE stain, differences in two groups of specimens of psychology, and carries on the classification according to the Pfirrmann grading standard, and use the mouse anti MMP-3 and MMP-9monoclonal antibody, using immunohistochemical method to detect intervertebral disc tissue expression in two groups of specimens and comparative analysis. Positive expression of indications for detect the tan or yellow are observed in the cytoplasm.Results: 1. The control group(normal lumbar nucleus pulposus tissue) : slice is visible in the extracellular matrix with cartilage cells and nucleus pulposus cells scattered distribution, clear contour, good coated structural integrity, cell morphology more short for polygon or fusiform, cytoplasm are evenly distributed, the nucleus of violet, and the number is more, the number of cartilage in the pit distribution more, collagen red, around a whole order rules, presents the network structure. Degeneration group(patients with lumbar disc prolapse of lumbar nucleus pulposus tissue) : slice shows cartilage cells and nucleus pulposus cells are evenly distributed, the less the number of cells distributed in cartilage in the pit, shape of spindle, and outline is not clear, the mitochondria in cell number is less, and lysosome gathers apparent, collagen tissue around not neat, to red, to present a sample cavity structure as a whole. 2. Immunohistochemical detection of HE staining results: control group, and organization of nucleus pulposus of lumbar disc degeneration group in MMP-3 and MMP-9 are distributed in the cytoplasm, immunohistochemical detection showed positive for lumbar nucleus pulposus tissue of MMP-3 and MMP-9 under a microscope, the nuclei are pale blue, and chondrocytes brown or tan can be seen in the cytoplasm. 2.1.Control nucleus pulposus cells positive expression of MMP-3 rate was(6.76 ±2.32) %,(27.85±7.52) % for outstanding group, hernia group(61.64±9.74) %,(76.52±6.72) % for free group, visible in the cell nucleus pulposus degeneration group of MMP- 3 positive expression rate was significantly higher than the control group, and had obvious statistical significance difference(P < 0.05), and the degeneration group in each subgroup(different types) in nucleus pulposus cells of MMP-3 positive expression rate also has significant statistical difference(P < 0.05).2.2.The control group in the cell nucleus pulposus positive expression of MMP-9 rate was(7.74±2. 36) %,(45.06±4.43) % for outstanding group, hernia group(81.43±3.41) %,(84.35±4.34) % for free. Visible in the cell nucleus pulposus degeneration group of MMP-9 positive expression rate was significantly higher than that of control group, and had obvious statistical significance difference(P < 0.05), while out and free group nucleus pulposus cells compared to positive expression rate of MMP-9, there was no significant difference statistically significant(P > 0.05). 2.3.Under Pfirrmann grading standard of class II is divided into three parts: the nucleus pulposus cells, the positive expression of MMP-3 rate was(8.78±2.62) %, III(33.47±6.79) %, IV for(51.24±10.59) %, V level(66.73±9.84) %, visible under the Pfirrmann grading standard, between the different levels of MMP-3 the positive expression rate of comparative differences have significant statistical significance(P < 0.05). 2.4.Under Pfirrmann grading standard of class II is divided into three parts: the nucleus pulposus cells, the positive expression of MMP-9 rate was(10.53±3.43) %, the level III for(52.47±10.42) %, level IV for(67.46±9.64) %, V level for(83.36±6.68) %, visible under the Pfirrmann grading standard, between the different levels of MMP-3 the positive expression rate of comparative differences have significant statistical significance(P < 0.05). 2.5.By Pearson correlation analysis results show that the degeneration of nucleus pulposus cells positive expression of MMP-3 and nucleus pulposus cells positive expression of MMP-9 was positively related to(0 < r < 1).Conclusions: 1. In the degeneration of intervertebral disc nucleus pulposus tissue MMP-3, the positive expression of MMP-9 is significantly higher than normal intervertebral disc tissue MMP-3, the positive expression of MMP-9, MMP-3 and MMP-9 expressed in lumbar organization change may participate in the process of organization of lumbar disc degeneration.2. MMP-3 and MMP-9 in degenerative lumbar nucleus pulposus tissue becomes clear positive correlation, speculated that MMP-9 May participate in the MMP-3 activation process. 3. In Pfirrmann grading, different levels of MMP-3 in lumbar disc degeneration, the difference of the positive expression of MMP-9 obviously, confirmed that the two can reflect the process of intervertebral disc degeneration. |
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