The Effect Of Shennao Fuyuan Decoction Combined With NSCs Transplantation On Caspase-3 And Caspase-9 In MCAO Rats

Objective:To investigate the effect of Shennao Fu yuan decoction combined with NSCs transplantation on Caspase-3 and Caspase-9 in MCAO rats.Methods:Healthy male SD rats of clean grade,weighing 260 ±20g,were used for one week of adaptive feeding.MCAOO models of middle cerebral artery obstruction in rats were established by thread embolization,except for sham operation group(simple operation without thread insertion,infusion of distilled water of the same volume).After the successful establishment of the model,the model group was randomly divided into three groups: model group(treated with MCAO,distilled water of equal volume by gastric instillation),traditional Chinese medicine group(treated with MCAO,intragastric administration of equal volume of kidney and brain and Fuyuan decoction),and combined group(with MCAO operation).Equal volume fetal rat NSCs was injected into the ischemic lateral ventricle by lateral ventricle approach and fed with equal volume of kidney and brain Fuyuan decoction.Each group was given each of them.At the time point,6 experimental animals were guaranteed to be successful,all of them were given intragastric administration once a day until death.The above four groups were killed at 1 day,3 days and 7 days after the successful administration of the drug,and the ar ea of cerebral infarction was measured by TTC staining.The expression of Caspase-3 and Caspase-9 in brain tissue was determined by WB and immunohistochemistry,and the neurological impairment score was evaluated by m-NSS at 2 h after model and before deat h(3 times in each group,the average value was included in the result.Results:1.The neurological function score of each group decreased at each time point and before treatment,and the difference was statistically significant(P < 0.05 or P < 0.01).The nerve function defect score of MCAO rats in the Chinese medicine group and the combined group was lower than that in the model group,the difference was significant(P < 0.05 or P < 0.01).The difference was statistically significant.The effect of the combined group on reducing the neurological deficit score was more obvious than that of the traditional Chinese medicine group(P < 0.01).2.The percentage of cerebral infarction area in each group was increased in the first three days except the sham operation group.At the same time point,the percentage of cerebral infarction area in the Chinese medicine group was lower than that in the model group(P < 0.05),in the combined group was lower than that in the model group(P < 0.01),and in the tradition al Chinese medicine group was lower than that in the combined group(P < 0.05).3.The results of immunohistochemistry showed that the expression of Caspase-3 and Caspase-9 in sham operation group was significantly lower than that in model group(P < 0.01).Compared with the model group,the Chinese medicine group was lower than the model group(P < 0.01),the combined group was lower than the model group(P < 0.01),and the combined group was lower than the traditional Chinese medicine group(P < 0.01).The expression of Caspase-3 and Caspase-9 was more obvious in the combined group.4.Detection of WB in sham operation group at each time point compared with the model group,the expression of only a small amount of Caspase-3,Caspase-9(P < 0.01),suggesting that the model was successful,which were caused by MCAO in brain tissue of rats with a large number of neurons apoptosis.Other groups on the third day the expression peak,rising three days before and three days after the downward trend and immunohistochemical results generally matched.In the same time,the Chinese medicine group,the expression of combination group was lower than that of model group(P < 0.01),Chinese medicine group group,combination group expression were lower(P < 0.01).Conclusion:Renal and cerebral rehabilitation combined with NSCs transplantation can improve the neurological deficit score,reduce the volume of cerebral infarction and inhibit the expression of Caspase-3 and Caspase-9 in MCAO rats after cerebral infarction.The effect is better than that of Shennao Fuyuan decoction alone.