| Objective:Adopting the method of MCAO(middle cerebral artery occlusion) to make model, establishing the rats infarction model, to observe the effect of sanqi panax notoginseng for injection on neurologic function score, protein level and mRNA level of nerve growth inhibitory factor Nogo-A and its receptors p75 and NGR at different time after operation.Finally discuss the cerebral function remodeling, reconstruction mechanism of xuesaitong in cerebral infarction recovery from protein level and gene level.Methods:Adult health male SD rats, weight 300±20g. Rats were randomly divided into operation and non-operation group (normal group).The operation group applied improved Longa method to make the focal and permarent model of MCAO, and implemented EZ-Longa evaluation after the rats woke up for 2 hours. According to the evaluation the rats included in the standards were randomly divided into model group,normal saline group, xuesaitong small doses group (3.6mg/100g. d), xuesaitong high-dose group(7.2mg/100g. d), positive control group-nimodipine group(1.44mg/100g. d). They were administrated 5 hours after operation, and implemented EZ-Longa evaluation on 7th,14th and 28th day after operation. Each group had been raised 7days,14days and 28days before executed, and then took the brain tissue depending on the cross 2mm before and after to dehydrate, embed in paraffin routinely, slice thickness about 4um continuous coronal plane,make paraffin section. Quickly brung out infarction brain tissue about 200mg, divide into 2 tubes in liquid nitrogen to preserve, finally the expression changes of the receptors of Nogo-A p75 and NGR would be observed by motheds of immunohistochemistry, western blot and Real-time PCR.Results:1 preparation of rats MCAO model and evaluation of neurologic functionThe neurologic function had been evaluated on 7th,14th and 28th day after MCAO operation. The normal group had no neurological function defect symptoms. The scores of model group were significantly higher than normal group (p<0.01or p<0.05) at different time after MCAO. With time prolonging the neurologic function scores of operation group had been gradually reduced, compared with the model group at the same time, the treatment group had more obvious trend,especially xuesaitong high-dose group and nimodipine group. Body mass index is a kind of monitoring indicator acting as evaluating overall health of the rats after MCAO. Respectively took and recorded weight on preoperative and 7th,14th and 28th day after MCAO operation. Took preoperative weight as basis value, body mass index namely for the ratio between the weight at different time and the basic value. The result showed:the body mass index of each group appeared chiefly decreased and then increased, especially the weight of treatment group back faster and more obvious.2 effect of sanqi panax notoginseng for injection on the protein expressions of the receptors of Nogo-A p75 and NGR at different time after MCAO (the method of immunohistochemistry)Adopting the method of immunohistochemistry, we oberseved effect of sanqi panax notoginseng for injection on the protein expressions of the receptors of Nogo-A p75 and NGR, in different infarction areas(infarction area, borderline area, and the normal region), at different time(7th,14th and 28th day) after MCAO. The results showed that:In the infarction area, with time prolonging the espression of NGR and p75 gradually reduced. Xuesaitong and nimodipine group were higher than the model group at the same time;In the borderline area, the espression of NGR and p75 of model group significantly increased than normal group. With time prolonging the espression of NGR gradually increased, and reached the high rate on 28th day,while the espression of p75 reached the high rate on 14th day, and slightly reduced on the 28th day. The treatment group were lower than the model group at the same time; In the normal region, the espression of NGR and p75 of model group increased than normal group,while treatment group reduced, there was no statistically significant difference among each group.3 effect of sanqi panax notoginseng for injection on the protein expressions of Nogo-A and its receptors p75 and NGR at different time after MCAO(the method of western blot)Adopting the method of western blot,we oberseved effect of sanqi panax notoginseng for injection on the protein expressions of Nogo-A and its receptors p75 and NGR at different time (7th,14th and 28th day) after MCAO from the protein level. The result showed:To the normal group at different time, the protein expressions of Nogo-A and its receptors p75 and NGR had no statistically significant difference. On the 7th day, the expression of Nogo-A and its receptor p75 of model group increased, reached the high rate on the 14th day, and slightly reduced on the 28th day. To NGR, the model group gradually increased with time prolonging, and reached the high rate on the 28th day. To the treatment group, the protein expressions of Nogo-A and its receptors p75 and NGR were lower than the model group,and gradually decreased with time prolonging, xuesaitong high-dose group lower than xuesaitong small doses group(P<0.05 or P<0.01).4 effect of sanqi panax notoginseng for injection on the mRNA expressions of the receptors of Nogo-A p75 and NGR at different time after MCAO(the method of Real-time PCR).Adopting the method of Real-time PCR,we oberseved effect of sanqi panax notoginseng for injection on the mRNA expressions of the receptors of Nogo-A p75 and NGR at different time after MCAO from the gene level. The results showed:To the normal group at different time, the mRNA expressions of Nogo-A and its receptors p75 and NGR had no statistically significant difference. To the expression of NGR, the model group gradually increased with time prolonging, and reached the high rate on the 28th day, while p75 increased on the 7th day, reached the high rate on the 14th day, and slightly reduced on the 28"'day. To the treatment group, the mRNA expressions of p75 and NGR were lower than the model group,and gradually decreased with time prolonging, xuesaitong high-dose group lower than xuesaitong small doses group(P<0.05).Conclusion:1 Sanqi panax notoginseng for injection can reduce neurologic deficiency, promote neurological function resume after cerebral ischemia;2 Sanqi panax notoginseng for injection can decrease the protein expressions of Nogo-A and its receptors NGR and p75 in different infarct ion areas at different time after MCAO;3 Sanqi panax notoginseng for injection can decrease the mRNA expressions of the receptors of Nogo-A p75 and NGR at different time after MCAO.4 Sanqi panax notoginseng for injection can decrease the protein and mRNA expressions of the receptors of Nogo-A p75 and NGR, which may be one of the remodeling mechanisms after MCAO. |
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