| Part Study on Xingnaojing Oral Liquid(XOL) treating the nerve functional damage and Ⅰthe inflammatory injury of neurons caused by cerebral ischemia. 1.Estimate the effect of XOL on neuropathic symptoms and Cerebral tissue pathomorphology of MCAO rats. Objective: Estimate the protective effect of XOL on acute damage of cerebral ischemia and reperfusion through observing symptoms of neurological deficit and the extent of cerebral infarction of rats. Method: Using suture-occluded method to build the model of Middle cerebral artery occlusion(MCAO) after rats having had drugs for 5 days, the symptoms of neurological deficit, the pathological morphology change of cerebral tissue and protective effect of XOL on damage of acute cerebral ischemia and reperfusion were estimated with 5grades scores of Zea Longa. Result: Remove the filament occlusion 2hours after the Cerebral ischemia and 22 hours, 70 hours after the reperfusion apart, rats in the sham operation group had no symptoms of neural functional defect, but other groups all appeared to have symptoms of neural functional defect more or less 22 hours as well as 70 hours after the Cerebral ischemia. At the same time, every medication administration group had different effect of improving symptoms of neurological deficit in rats. The Beam-walking experiment showed that comparing with the model group, operation group showed significant difference(P<0.05), the sham operation group showed to be different as well, while the XOL group(50mg/kg) showed to be different in a certain way. 22 hours, 70 hours after the Cerebral ischemia reperfusion, there came some morphologic changes in pallium and hippocampal tissues, local edema and inflammatory reaction, obvious local pale, neuron ranked few and far between and large amount of whom became necrotic tissues, XOL could promote these situations in certain extents. Conclusion: 22 hours and 70 hours after the rats had suffered acute Cerebral ischemia reperfusion, model group had obvious symptoms of neurological deficit, all XOL groups could improve neurological deficit in different degrees and reduce the necrosis ranges.2.Effects of XOL on activity of NSE and content of S100 B in blood serum of MCAO rats.Objective: Study how the drugs effect the neurocytes and the injury degree of neurocytes after Cerebral ischemia and reperfusion through testing activity of NSE and content of S100 B in blood Serum of MCAO rats with nerve damaged. Method: Copy rats MCAO models with suture-occluded method after rats having had drugs for 5 days, remove the filament occlusion 2hours after the Cerebral ischemia and execute the rats 22 hours, 70 hours after the reperfusion respectively, test the activity of NSE and content of S100 B in blood serum with enzyme-linked immunosorbent assay(ELISA). Result: Remove the filament occlusion 2hours after the Cerebral ischemia and 22 hours, 70 hours after the reperfusion apart, comparing with the model group at the same time, activity of NSE and content of S100 B in rats blood serum was improved in different degrees. Medication administration groups showed a depressing(P<0.1) in activity of NSE 22 hours and different degrees of depressing in content of NSE 70 hours after reperfusion. Comparing with the model group at the same time, XOL group(50mg/kg) and Zileuton group showed significant depressing in content of S100 B 22hours after reperfusion, XOL group(50mg/kg) group showed obvious depressing(P<0.05) in content of S100 B 70hours after reperfusion. Conclusion: There was promoting in content of S100 B and NSE in MCAO rats blood serum, then there was depressing in activity of NSE and content of S100 B in rats blood serum of XOL group, what suggested that XOL could be helpful in mitigating damage of the Cerebral ischemia through lowering the activity of NSE and content of S100 B in rats blood serum.3.Effect of XOL in content of inflammatory factor IL-8 and activity of MPO in MCAO rats’ cerebral homogenate. Objective: Detect the content of IL-8 and activity of MPO in MCAO rats cerebral homogenate, check out the changes in Neutrophil function and activity and cytokines, and investigate the depressing effect of XOL on experimental Cerebral ischemia and reperfusion. Method: Copy rats MCAO models by suture-occluded method after they having had drugs for 5days, remove the filament occlusion 2hours after the Cerebral ischemia, execute rats and test the content of IL-8 and activity of MPO in rats 22 hours and 70 hours respectively after the reperfusion with ELISA. Result: ①22hours after the reperfusion, comparing with the model group, sham-operation groups showed an obvious depressing(P<0.05) in activity of MPO in cerebral homogenate of the disease half brain; groups having drugs all showed depressing in activity of MPO in cerebral homogenate in a certain extent(P<0.05). There was depressing(P<0.1) in activity of MPO of XOL(50mg/kg, 25mg/kg)groups and Zileuton group 70 hours after reperfusion comparing with modelgroup. ②22hours after reperfusion, there was improving in content of IL-8 in cerebral homogenate of the disease half brain of rats in model group, comparing with which the sham-operation group, XOL(50mg/kg, 25mg/kg)groups and Zileuton group all showed obvious depressing in content of IL-8. Conclusion: There was obvious improving in activity of MPO and content of IL-8 in the disease half brain of MCAO rats, there was obvious depressing(P<0.1 or P<0.05) of Neutrophil function and activity and content of cytokines in XOL group, which suggested XOL could reduce damage caused by Cerebral ischemia through monitoring the activity of MPO and content of IL-8.PartⅡEffect of XOL in regulating the pathways of leukotrienes and 5-Lox in MCAO rats brain tissues. 4.Effect of XOL on contents of Cys LTs, LTD4, LTB4, 5-Lox in MCAO rats brain tissues. Objective: Expect to find changes of contents of Cys LTs, LTD4, LTB4 and 5-Lox in MCAO rats brain tissues and estimate the curative effect of XOL through detecting changes of contents of Cys LTs, LTD4, LTB4 and 5-Lox in rats brain tissues 2hours after the cerebral ischemia, 22 hours and 70 hours after the reperfusion respectively. Method: Copy SD rats MCAO models by suture-occluded method after rats having had drugs for 5days, remove the filament occlusion 2hours after the Cerebral ischemia, execute rats and test the content of Cys LTs, LTD4, LTB4 and 5-Lox in rats’ brain tissues 22 hours and 70 hours after the reperfusion with ELISA. Result: ①22hours after the reperfusion, comparing with the sham-operation group, there was obvious improving(P<0.05) in content of 5-Lox in rats cerebral homogenate of model group and there was improving in content of 5-Lox in rats cerebral homogenate at 70 hours as well, there had no difference comparing 22 hours with 70 hours after the reperfusion in model groups; comparing with model group, XOL group(25mg/kg) showed an obvious depressing(P<0.05) 22 hours after the reperfusion, Zileuton group showed a significant depressing(P<0.01). Zileuton group showed depressing as well 70 hours after the reperfusion. ②22hours after the reperfusion there was improving(P<0.1) in content of LTB4 in rats cerebral homogenate of model group comparing with sham-operation group, comparing with model group XOL group(25mg/kg) as well as Zileuton group both showed depressing(P<0.1) in content of LTB4 in rats cerebral homogenate; 70 hours after the reperfusion there was improving(P<0.1) in content of LTB4 in rats cerebral homogenate of model group comparing with sham-operation group, comparing with model group Zileuton group showed depressing(P<0.1) in content of LTB4 in rats cerebral homogenate, there had no difference in model groups comparing 22 hours with 70 hours after the reperfusion.③22hours after the reperfusion there was improving(P<0.1) in content of LTD4 in rats cerebral homogenate of model group comparing with sham-operation group, comparing with model group the medication administration groups all showed obvious depressing(P<0.05) in content of LTD4 in rats cerebral homogenate; 70 hours after the reperfusion there was obvious improving(P<0.05) in content of LTD4 in rats cerebral homogenate of model group comparing with sham-operation group, comparing with model group Zileuton group showed obvious depressing(P<0.05) in content of LTD4 in rats cerebral homogenate, there had no difference comparing 22 hours with 70 hours after the reperfusion in model group. ④22hours after the reperfusion there was significant improving(P<0.01) in content of Cys LTsin rats cerebral homogenate of model group comparing with sham-operation group, compering with model group XOL group(50mg/kg) showed obvious depressing(P<0.05) while XOL group(25mg/kg) and Zileuton group showed depressing(P<0.1) in content of Cys LTsin rats cerebral homogenate; 70 hours after the reperfusion there was significant improving(P<0.01) in content of Cys LTsin rats cerebral homogenate of model group comparing with sham-operation group, comparing with model group XOL group(50mg/kg) as well as Zileuton group both showed depressing in certain extents(P<0.1 or P<0.05) in content of Cys LTs in rats cerebral homogenate, there had no difference comparing 22 hours with 70 hours after the reperfusion in model group. Conclusion: There were improving in contents of Cys LTs, LTD4, LTB4 and 5-Lox in certain extents in MCAO rats cerebral homogenate of model group. All medication administration groups reduced contents of Cys LTs, LTD4, LTB4 and 5-Lox in certain extents in rats cerebral homogenate, which suggested that XOL may have curative effect on Cerebral ischemia and reperfusion through restraining the pathway of arachidonic acid-5-Lox.5.Effect of XOL in intervening proteins expressions of Cys LT1 and CysLT2 in MCAO rats cerebral tissues. Objective: Expect to discover the target on which XOL works to fix inflammatory injury caused by cerebral ischemi through detecting proteins expressions of Cys LT1 and Cys LT2 in rats cerebral tissues harmed by experimental Cerebral ischemia and reperfusion. Method: Copy SD rats MCAO models by suture-occluded method after they having had drugs for 5days, remove the filament occlusion 2hours after the Cerebral ischemia to give reperfusion to rats. Test protein expressions of Cys LT1 and Cys LT2 in rats pallium and hippocampus with western blotting 22 hours and 70 hours after the reperfusion respectively. Result: 22 hours after the reperfusion, rats from model group expresses more proteinCys LT1 and Cys LT2 in certain extent(P<0.01 and P<0.05) than those from the sham-operation group, each dose group of XOL could lower proteins expressions of Cys LT1 and Cys LT2 in rats pallium and hippocampus. Conclusion: Proteins Cys LT1 and Cys LT2 express obviously more in disease half pallium and hippocampus of MCAO rats, Cys LT1 and Cys LT2 may be the targets on which XOL works to regulate and control inflammatory injury caused by cerebral ischemi. Conclusion: XOL could reduce Neuropathic injury in MCAO rats cerebral tissues. Related links include: ①Restrain the activity of NSE and content of S100 B, ②Monitor the inflammatory factors and activity of MPO, ③Restrain the content of Cys LTs, LTD4, LTB4, 5-Lox and the proteins expressions of related receptors such as Cys LT1 and Cys LT2. Cys LT1 and Cys LT2 may be the targets on which XOL works to regulate and control inflammatorymetabolic disturbance to get the point of preventing and treating cerebral damage of Cerebral ischemia and reperfusion. |
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