| Purpose:Exploring the protective effect and mechanism of Danshen and Tanshinone during rat spinal cord ischemia-reperfusion injury by observing the spinal cord tissue in water content,Ca2+centration, cytochrome C, caspase-3 and influence of pathological in spinal cord tissue, in order to provide new ideas and experimental basis for clinical prevention and treatment of spinal cord ischemia-reperfusion injury.Methods:①A total of 160 Sprague Dawley rats, weighing (270±10)g,of clean grade and either gender, were provided by Shanghai Slaccas. After adaptable feeding for a week, by adopting the improvement of ZHANG LI et al[1].the rats were randomly assigned to:sham-surgery (A group n=40), ischemia-reperfusion (Bgroup n=40), Danshen intervention (Cgroup n=40), and Tantanshinone intervention groups (Dgroup n=40).we prepare the model of the rat spinal cord ischemia-reperfusion injury:Danshen intervention group and Tanshinone intervention group:half an hour before surgery by intraperitoneal injection of Danshen, Tanshinone Ha sodium injection; ischemia-reperfusion group (Bgroup):no injection of any medication, the others are the same with the two groups; sham-operation group(Cgroup), anesthesia and surgical operation with the previous three groups, but no occlusion of the abdominal aorta or the termination of surgery, at room temperature, the natural wake. After modeling, at 0.5h, Ih, 4h,8h,12h, take out L2-L5 spinal cord.②Calculating the changes in spinal cord tissue water content, atomic absorption spect-rometry detection of Ca2+concentration changes by the dry method, immunohisto chemical detection of spinal cord nerve cells of Cytochrome C release, RT-PCR detection of spinal cord nerve cells in Caspase-3 protease expression and HE staining under light microscope observation of spinal cord tissue pathological changes.③Using spss15.0 statistical analysis software and image processing system,we collect, classify and analyze the data and image.Results:①Using ZHANG Li, et al. Improvements, we produce the model of the Act of spinal cord ischemia-reperfusion injury that is ischemia complete, reproducible, easy to operate, and economical and practical. ②The water content and Ca2+concentration in the sham-operated group of spinal cord tissue have significant change. There is no cytochrome C release in spinal cord nerve cells, no expression in caspase-3, The observations compared between sham-operated group and Danshen, Tanshinone intervention group at each time point have significant difference. It was statistically significant(p<0.05).The expression of Caspase-3 in ischemia reperfusion group significantly increased, the release of cytochrome C increased significantly and produced large amounts of Ca2+, ischemia-reperfusion injury in spinal cord tissue edema, the water content could be up to 74.51%, ischemia-reperfusion group compared with the sham-operati-on group at the same time has very significant differences (P<0.01),compared with Danshen intervention group or Tantanshinone intervention group at the same time has significant differences(P<0.05);The Caspase-3 expression in Danshen and Tanshinone intervention group was significantly reduced, the release of cytochrome C was significantly inhibited, int-racellular Ca2+content significantly reduced, spinal cord edema has been significantly improved, water content decreased significantly, the observations compared between sham operation group and ischemia-reperfusion group at same time points have very significant difference (P＜0.05).Conclusion:①According to ZHANG Li et al improvements, the model of spinal cord ischemia-rep-erfusion injury, which has good repeatability, complete ischemia, easy to operate, is the ideal model to study the spinal cord ischemia-reperfusion injury, worthy of clinical application.②In the spinal cord ischemia-reperfusion injury, Ca2+, cytochrome C and Caspase-3 were involved the pathological process.Danshen and Tanshinone can significantly reduce the ischemia-reperfusion injury in the spinal cord caused by spinal cord edema, maintain the intracellular calcium balance, reduce mitochondrial release of cytochrome C, and inhibit the expression of caspase-3, thereby having a very good protective effect on spinal cord ischem-ia-reperfusion injury.It may be related to reduce water content in spinal cord and Ca+ Centration, reduce mitochondrial release of cytochrome C, and inhibit the expression of caspase-3.③Confirmed by morphology and Danshen or Tanshinone can really ischemia-reperfus-ion injury of the spinal cord play a protective role. |
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