| Intracerebral hemorrhage is a complex disease which has the features of urgent attaching, fast change, critical condition, and it accounts for about10%-25%of the cerebrovascular disease,70%-80%of the morbidity,38%-43%of the mortality,1.8%-11%of the recurrence rate. Intracerebral hemorrhage not only causes serious influence on the patients, and also brings heavy burden to the society and family, which must be treated emergently to save lives and improve the quality of life. After Intracerebral hemorrhage, the brain tissue damage is directly derived from the hematoma mechanical oppression, and the main damage is secondary brain injury, brain edema and inflammatory reaction which are the main culprit of secondary brain injury. Thrombin occupies an important position in the secondary injury, and NF-KB, MMP-9and NMDA receptors play an important role in the secondary brain damage. The mechanism of brain nerve damage in this disease has not been fully elucidated, and the treatment also lack of breakthrough. Most patients has neurological functional defect. TCM has its advantages and from the perspective of the poison and blood stasis to explore its mechanism. Panax Notoginseng Saponins is extracted from Chinese herb pseudo-ginseng, which has the effect of reducing brain edema and inflammatory reaction, promoting the absorption of cerebral hemorrhage of intracranial hematoma.Objective:This study used experimental animals to create intracerebral hemorrhage model, and selected Chinese medicine notoginseng extracts Panax Notoginseng Saponins to observe the nerve function defect change, brain tissue morphology changes by examining the brain water content and immunohistochemical experimental and the protein and mRNA expression by the western blot and PCR methods, to explore the mechanism of Panax Notoginseng Saponins on the effects of secondary brain injury and brain protection.Methods:Healthy homologous male SD rats,230-250g, were randomly divided into control group, model group and treatment group, and observe6h,24h,48h,72h and7days respectively. Using rat stereotaxic instrument and microsyringe to inject2uL â…£ collagenase into the caudate putamen (model group and treatment group), while the control group was injected saline instead. Saline and Panax Notoginseng Saponins were treated by intraperitoneal injection.6hours time points, All groups were treated immediately after surgery and the dose depend on the weight. The rats were treated everyday and acess the nerve function score, and the brain water content by Billiot formula, and the protein and mRNA expression by the western blot and PCR methods at each time point. Experimental data were analyzed by the single factor analysis of variance (One-Way ANOVA) in SPSS17.0statistical software package, and P<0.05as the boundaries that the difference was significant.Results:1. Neural function defect scale:there is a significantly difference between model group and control group. mNSS score in the model group was significantly higher, from6h to48h, and increased to peak at48h and then began to decrease slowly. In the treatment group, the score was significantly decreased comparing with the model group at movement, sensation, balance and reflect nerve function at each time point except at6h time point(P=0.228).2. Brain water content:the brain water content in the model group showed a trend of increase first at6time point, and peaked at48h, and declined from72h and then back to the normal level and there is significantly difference at each time point(P<0.05). Cerebral edema has been reduced in the treatment group at each time point and there is a significant difference at24h,48h, and72h time points comparing with the model group,(p<0.05). At6h and7days time points, the difference are not significant, medication group compared with model group, while reduced cerebral edema, but there was no statistical difference (p=0.062and p=0.41respectively).3. HE and immunohistochemitry:It showed that the cell morphology is normal, neatly, without edema and inflammatory cells in caudate putamen of the sham group by the HE staining. In the model group, the nerve cells are swelling; the gap between the cells increases, and the cell begin pycnosis and necrosis, beginning from6h, and peaking at48h and then gradually decrease. Immunohistochemical results:the expression of PAR-1, NMDA, NF-KB and MMP-9in the model group showed a trend of increase first at6time point, and peaked at48h, and then declined and there is significantly difference between model group and sham group at each time point(P<0.05). the expression of PAR-1, NMDA, NF-KB and MMP-9reduced in the treatment group at each time point and there is a significant difference at each time points comparing with the model group (p<0.05).4. Western Blot test:the expression of PAR-1in the model group showed a trend of increase first at6time point, and peaked at48h, and then declined and there is significantly difference between model group and sham group at each time point except at6h time point(p=0.074). The expression of NMDA reduced in the treatment group at each time point and there is a significant difference at each time points comparing with the model group except at6h(p=0.131). The expression of NMDA in the model group showed a trend of increase first at6time point, and peaked at48h, and then declined and there is significantly difference between model group and sham group at each time point. The expression of NMDA reduced in the treatment group at each time point and there is a significant difference at each time points comparing with the model group except at6h(p=0.072) and7d time point(p=0.145).5. Rt-pcr test:the expression of PAR-1in the model group showed a trend of increase first at6time point, and peaked at48h, and then declined and there is significantly difference between model group and sham group at each time point. The expression of PAR-1reduced in the treatment group at each time point and there is a significant difference at each time points comparing with the model group. The expression of NMDA in the model group showed a trend of increase first at6time point, and peaked at48h, and then declined and there is significantly difference between model group and sham group at each time point. The expression of NMDA reduced in the treatment group at each time point and there is a significant difference at each time points comparing with the model group. The expression of NF-KB in the model group showed a trend of increase first at6time point, and peaked at48h, and then declined and there is significantly difference between model group and sham group at each time point. The expression of NF-KB reduced in the treatment group at each time point and there is a significant difference at each time points comparing with the model group. The expression of MMP-9in the model group showed a trend of increase first at6time point, and peaked at48h, and then declined and there is significantly difference between model group and sham group at each time point. The expression of MMP-9reduced in the treatment group at each time point and there is a significant difference at each time points comparing with the model group.Conclusion:1. Panax Notoginseng Saponins promotes the recovery of neurologic deficits in rats after cerebral hemorrhage.2. The abnormal expression of NMD A, PAR-1, NF-KB and MMP-9play important roles in secondary brain injury after intracerebral hemorrhage.3. Panax Notoginseng Saponins has a decrease effect on the expression of PAR-1, NMDA, NF-KB and MMP-9.4. Panax Notoginseng Saponins has a protection effect after cerebral hemorrhage in rats. |
