Expression And Significance Of Caspase-3, IL-1 β And C-fos In The Cervical Cord And Stomach Of Rats With Cervical Spondylosis

Study Background: (1)Anatomic foundation (1)sympathetic nerve is distributed in Luschka , articular capsule, cervical miniarticular, posterior longitudinal ligament, posterior annulus fibrosus of intervertebral disk, neuronal periphery of dorsal root ganglion and arteriae vertebralis. Other enthetic nerves interreact with dorsal root ganglion by non-synapsis. (2)Both with sensation and movement, gastrointestinal tract is the only organ dominanted by central nervous system, enteric nervous system and autonomic nerve system.(3) Need successful models ofcervical spondylosis.(2)Advancement Neurogastroenterology was proposed in Rome II system in 1999,the core was pathogenic mechanism of gastrointestinal dysfunction, clinical manifestation and ethological formation of living being-society- psychology. The new classification of the World Gastroenteropathy Meeting in Bangkok in February 2002, Neurogastroenterology is neurology of motive disease, researching function and dysfunction of digestive tract, function and dysfunction and structural abnomalities of brain and cord, domination of sympathesis, parasympathesis and autonomic nerve of intestine and the interaction between centrol nerve system and nerves of digestive tract.(3) Experiment foundation IL-1 β is an important inflammatory cytokine. Caspase-3 is one of key members of interleukin 1β convertiny enzyme family. Not only in acute injury and but also in chronic degeneration, IL-1β induced neuron apoptosis of centrol nerve system through activating Caspase-3, and Caspase-3 is a potential mediator in apoptosis. Regulated by IL-1 β , c-fos is expressed as third messenger control ing target gene.Objective: To establish cervical spondylosis of muscle imbalance and posteriorcolumn instability in rats, then study expression of Caspase-3, IL-1β and c-fos in cervical cord and stomach of the rats with cervical spondylosis, and the co-expression of them in cervical cord and stomach. Analyze the relationship among them and study the mechanism of neck-stomach syndrome, desire to explain the unknown pathogenesy of functional dyspepsia (FD) and irritable bowel syndrome( IBS) from spondylotic speciality.Methods: Sixty SD rats (four months old) were randomly divided into control, muscle imbalance and posterior column instability groups which were subdivided into groups of 2 and 4 months, ten rats in each group. The X-ray films and motion function of oblique board test were evaluated. C3-7 discs were collected, dealt with decalcification and dehydration, then with paraffin imbedding, sagittal microtome section and HE dyeing. Study morphologic changes of disc according to Miyanmoto methods. Other 96 SD rats were destroyed the stability of cervical posterior column and established cervical spondylosis models, the cord segments C₄-6 and stomaches were collected 3 months and 5 months after the operation. Rats with sham operation were used as controls. Determined the expression of Caspase-3 and IL—1 in cervical cord by immunohistochemistry and Western Blot methods, Countered the positive cells under microscope of 200 times at random with 10 fields, then the data were analyzed statistically. The results of densitometry readings of gel bands were expressed as arbitrary units. The changes of c-fos expression in cord andgastric myenteric plexus were analyzed by immunohistochemistry methods. Results: In the control group, the J-ray films showed nature curve, normal vertebralclearance, no osteosis spur and ossify in Luschka joint and joint process. But in muscle imbalance and posterior column instability groups, showed disappeared or stiff nature curve, stenosis vertebral clearance, osteosis spur formed, ossify or sclerosis emerged in Luschka joint and joint process. Annulus fibrosus aligns acceptable in degenerated normally discs, light hyaline degeneration in it under microscope, there was significant difference in the grade score according to Miyanmoto standard. Compared with control group in oblique board test, the muscle force decreased in muscle imbalance and posterior column instability groups. Therewas no statistic significant difference between control groups of 2 months and 4 months, and also in muscle imbalance and posterior column instability groups, but there was significant difference in muscle imbalance and posterior column groups of 2 months, 4 months. The expression of Caspase-3 and IL-1P determined by immunohistochemistry methods in spinal cord, there was significant difference in control and model groups, and also in 3 months and 5 months model groups (P < 0. 05). The expression of Caspase-3 and IL-1 P determined by immunohistochemistry methods in stomach, there was significant difference in control and model groups (P < 0. 05 ) , and no significant difference in 3 month and 5 month model groups (P > 0.05) . Western Blot analysis showed time-dependent changes of Caspase-3 and IL-1 {3 protein in the cervical cord of rats with cervical spondylosis. Sample from control and model cords were prepared for Western Blotting at 3 months and 5 months. There was no significant expression of Caspase-3 and IL-13 protein in the control groups of 3 months and 5 months after sham operation. However, there was significant different expression between rats kept for 3 months and those for 5 months in the model groups (P < 0.05 ). There were a little c-fos positive neurons in cervical cord and gastric myenteric plexus by immunocytochemical analysis in control groups, More importantly, there was significant difference in c-fos coexpression between rats followed up for 3 months and those for 5 months in the model group ( cervical cord: 11.20 + 2.26, vs 27.68 + 4.36, P < 0. 05 ) ; and gastric antrum: 11.3 + 2.3, vs. 29.3 + 4.6, P < 0.05 ). There was no significant difference between rats followed up for 3 months and those for 5 months in the control group.Conclusion: In this study, Cervical posterior column instability and muscle imbalance can all result in cervical spondylosis, both muscle force balance and posterior column were important in maintaining cervical stability. We regarded c-fos regulated by IL-1 3 in vivo and confirmed that the simultaneous c-fos expressions in cervical spinal cord and gastric myenteric plexus were significantly different in the cervical spondylosis groups at 3 months and 5 months after operation. Therefore, there is a relationship between the neck and stomach, andthe gastrointestinal function may be affected by cervical spondylosis, if this hypothesis is confirmed by further studies, functional gastrointestinal diseases such as functional dyspepsia and irritable bowel syndrome could be explained by neurogastroenterology.