The Expression of Proinflammatory Cytokines at the Early Stage of Cervical Spondylotic Myelopathy
|School||Hebei Medical University|
|Keywords||cervical vertebrae intervertebral disk tumor necrosis factor interleukin-1 interleukin-6 stability spine|
Objective: As everyone knows that the cervical spondylotic, CS is for short. The syndrome of the CS varies from one patient to another. And the classification system also presents a more and more varieties. According to the causes of the cervical spondylotic, we can have it as myelopathy, nerve root type, vertebral artery type, and mixed type. The myelopathy cervical spondylotic is the disease which caused by the herniated discs or the bone crest and have the myelopathy compressed and some related symptoms and signs. The most common sign is the weakness of the hands and legs. The feeling of the legs were abnormal, walking with instability, can’t move so fast, the hand grasp force weaken, easy to drop when holding the objects, paralysis of the both legs and hands. Weakness of the legs are also very common, in some severe patients we can also find some difficulties in the walking, urinary and fecal incontinence or urine retention, paralysis, et al. This type of cervical diseases has some difference between the other types, the mainly one is the radiation pain which can be common in the nerve root type, while it is not so common in other types. The common known causes to the cervical or the myelopathy cervical disease was that the degenerative changes of the cervical spondyle and the bone osteophyte formation in the back part of the cervical spine, or some other type of disc herniation, causing the myelopathy be compresed. Some researchers noted that the cytokines which produced in the procedures of the cervical spondylotic can be made this disease much more difficult to treat and distinguished from others. Others concluded that these cytokines and its related effects could cause the nerve root have the inflammatory changes or the level of the spondylotic, avagulate the symptoms of the patients, but the more details of this result was not known yet. The recent result shows a related relationship among the TNF-α、IL-1β、IL-6 and so on will have an effect on mediating the inflammatory reactions. These cytokines can be found a high expression of the cytokines, these are the common known as“Proinflammatory cytokines”. Among all these cytokines the TNF-αis the one which have the most important role in the reactions. With more and more research result come into been, the work effect have been known to both the doctors and the researchers. These days some drugs regarding to the TNF-α,the single-clone antibody Infliximab, have got the permission to be used in the clinical daily use, which was authorized by the Food and Drug Admission(FDA), the common suitable situation was the Rheumatic Arthritis, and so on. At present, there are some researchers want to present this kind of medicine in the treating of the disease such as disc degeneration, and some important results shows the good results of this kind of disease treating with the cytokines drug can be effectiveness. In our test, we want to discuss the expression change of proinflammatory cytokines at the early stage of cervical spondylotic myelopathy and the relationship with early disc degeneration. And make clear of the relationship between the clinical diagnosis, treatment both in the animal model or the clinical use. This research contains two parts, the first one is the animal model tests, while the other part of the research is the clinical case reports, and I will give a description below:The first part: animal test.Methods:we choose SD rat, in the back part of the cervical disc, we choose to plant the BMP, which can be used to introduce the disc and its related ligament calcification, the bone formation in the back part of the bone, and which can cause some symptoms of the cervical compression. In this test, we choose 36 SD rats, and separate them into the 2 groups randomly. Group A, 18 rats, the animal model using the method mentioned above. Group B, the contrast group, contains the same number of the same kind of rats, which was feed the same and living in the same environment as the testing group. We choose to kill the rats at the time point of 4, 8 and 16 weeks post-operation. Using the immune-history techniques to analysis the two group rats, and using these kinds of techniques to quantities the relations and the groups.Results:1. In group A, the animals showed that the moving ability was decreased in 8 weeks, 4 weeks post operation, we could find that the bone matrix was forming, which occupies the vertebral canal, and a lot of symptoms caused by the spinal cord compression and demyelination, or neuron deficit changes; which suggested the at the time point of 4 weeks, the myelopathy cervical spondylotic was formed, and at the time point of 8 weeks, this kind of disease was formed completely.2. The immune-histology results showed that the TNF-αwas mainly separated by the fibroblast,chondrocyte and lymphocyte, and at 4 weeks post-operation, there would not be that high level expression, which suggested the myelopathy cervical disease model was formed in the effects of the inflammatory as well as the cytokines. The quantities analysis of this results showed the secreting rates reached the highest points. TNF-αpositive number was 4, 6, 6 separately at the time point of 4,8 and 16 weeks. The immune-histology stain shows the TNF-αhas some relationships with the disc degeneration and its content level. It can give us a more detailed information this kind of cytokines were playing an important role in the inflammatory reaction.3. IL-1βalso lived mainly on the fibroblasts , chondrocyte and lymphocytes. But the difference was that it lived mainly on the lymphocytes. Nearly at 4 weeks this kind of the cytokines contained a much less level than the TNF-α, which showed a high levels in the 8 weeks or 16 weeks post-operation.4. IL-6 can play an important role in the reaction, which can cause the systematic symptoms. There were differences between the two groups, and the statistics difference existed. The second part:Clinical Case Study.Methods:we choose the patients information in the daily work experience in the 3rd hospital of Hebei Medical University. Thirty-two discs from 20 patients of cervical spondylotic myelopathy with course less than one month treated from May 2004 to January 2005 were collected and studied as experiment group, these patients all have a disease history less than 1 months, which is to say the patients will have a treatment both congenital or surgical interaction by the surgeon in less than one month, since the first time he or she got the uncomfortable feelings. The special one person, who performing this kind of work underguide by the same rules, harvested all the surgical spice man. And during this period 15 cervical discs from fresh cadavers with death time less than 24 hours were also collected and studied as control group. We collected the disc which contains both in these two groups. We choose to got the double-blinded principles, and have the 4% poly-paraformaldehyde; then according to the immuno-histochemistology stain to examine the cytokines contents, The expression of TNF-α, IL-1βand IL-6 was tested by immuno-histochemistology. Using the image procedure software to do the results, compare each of these groups whether the differences exists.Results:1. In the experiment group, the positive cases for TNF-α, were 15, In the control group, there were less positive cells in all the 15 cases, the positive cells shows a very limited number of cells. There were differences between the two groups by using SPSS 11. 5 statistics software (P<0. 05).2. In the experiment group, the positive cases for IL-1β, were12, In the control group, there were less positive cells in all the 15 cases, the positive cells shows a very limited number of cells. There were differences between the two groups by using SPSS 11. 5 statistics software (P<0. 05).3. In the experiment group, the positive cases for IL-6, were 11, In the control group, there were less positive cells in all the 15 cases, the positive cells shows a very limited number of cells. There were differences between the two groups by using SPSS 11. 5 statistics software (P<0. 05).4. In the experiment group, the positive cases for TNF-α, IL-1β, IL-6 were both positive, and among them all there were 12 patients which shows a very positive results in all the 3 groups. While in the control group the positive cells shows a very limited number of cells. There were differences between the two groups by using SPSS 11. 5 statistics software (P<0. 05).5. The results of the immune-chemistry showed a very good statistics differences exists among all the groups. Conclusion:The animal model of myelopathy cervical spondylotic can be reached by injecting the BMP at the disc level and the back part of it.The tissues of herniated disc can produce TNF-α, IL-1βand IL-6 cytokines which are mainly found in fibroblasts, cartilage cells and lymphocytes. These cytokines may play a role in the degeneration of cervical disc.