| OBJECTIVE: According to discuss the expression variation of IL-1α and BMP-7 in lumbar ligament flavum, we explore the function of them in the mechanisms of ligamentum flavum hytertrophy, which may provide a new treatment thought for lumbar canal stenosis caused by ligamentum flavum hytertrophy and ligamentum flavum ossification.METHODS:1. From January 2013 to October 2014, twenty specimens of ligamentum flavum were collected after surgery from Orthopedics Department of The Affiliated Hospital of North Sichuan Medicial college and were divided into two groups, the experimental group and control group. The patients in control group have some common characteristics: lumbar vertebrae fracture, aged between 18 and 30, male, having no postive past medical history, excluded from other chronic diseases, no ligamentum flavum hytertrophy in medical imaging data. In experimental group, patients were diagnosed with lumbar canal stenosis. They have characteristics as follows: 4 male and 6 female aged between 55 and 65, enjoying good health in the past, excluded from other chronic diseases, the thickness of hyperplastic lumbar ligament flavum of these poeple is more than 5 mm in medical imaging data. The process above has been approved by the ethics committee and been permitted by patients themselves and their families.2.After having been taken from vertebral plate slices, the specimens were embedded in conventional dehydration of paraffin after fixed in 10 percent neutral formaldehyde, serial section stored in freezer with 4℃,waiting for HE stain and immunohistochemical detection. Then, the specimens were saved in PCR fixed liquid and western blot fixed liquid in freezer with 4 ℃, waiting for the fluorescence quantitative PCR detection later. The experimental measurement data is expressed with mean standard deviation and analyzed with SPSS16.0 statistical software. The comparsion of every two mean adopts t test,Which is thought to have statistical signification about the difference when P is less than 0.05.RESULT:1. The comparison of specimens after surgery: the ones in control group are bright, soft, having smooth surface, with a clear boundary to surrounding tissue to be detached easily, fast retraction, curly with a good elasticity. However, the ones in experimental group are dark, hard, with rough surface, adhesion to adjacent tissue, hard to be detached. Comparing the ligamentum flavum without calcification and ossification in the control group with the ones in experimental group, the thickness of ligamentum flavum in latter is thicker, with statistical difference(P<0.05)。2.HE stain:The size of ligamentum flavum, arranged according to the long axis, is similar in control group and the surface of ligamentum flavum is smooth due to the thin and wavy elastic fibers paralleled with ligamentum flavum long axis.The fibers also has branches connected to form the reticular structure with a small amount of collagen fibers among it. On the other hand,the diameter of ligamentum flavum ranked disorderly is different. Fiber breaks with the increase of collagen fiber and structural deformation, which leads to the increace of fibroblast. Ossification or calcification is not detached. Comparing the amount of fibroblast in experimental group with which in control group, fibroblast cell number in former is obviously enhanced, with statistical difference(P<0.05).3. Immunocytochemistry: IL-1α postive cells in control group and in experimental group are 40%and 90% respectively, while BMP-7 postive cells in control group and in experimental group are 40%and 90% severally. The result above demonstrates that the expression of IL-1α and BMP-7 in experimental group is apparently higher than that in control group(P<0.05).4. Real-time fluorescent quantitative PCR detection: The gene amplification and melting curve are favourable. It is found with lumbar ligament flavum histological research that the expression of IL-1α and BMP-7 in experimental group is apparently higher than that in control group(P<0.05).5. Western blot test: the expression degree of IL-1α and BMP-7 in experimental group is apparently higher than that in control group(P<0.05).CONCLUSION:1. The enhancement of IL-1α expression may be one of the important mechanisms of ligamentum flavum degeneration.2. The enhancement of BMP-7 expression may also be one of the important mechanisms of ligamentum flavum degeneration. |
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