| BackgroundIschemic cerebral vascular disease(Ischemic cerebral vascular disease ICVD)under the category of "stroke" in traditional Chinese medicine is a clinical common acute critical illness.With the aging of the population in our country,the stroke incidence also increased significantly,due to diet,lifestyle changes,the proportion of young people with stroke also increased.The incidence of cerebral infarction and atherosclerosis,arterial atherosclerotic hard plaque Hyperplasia,breakage,emboli cause serious stenosis or occlusion of the vascular lumen,localized brain dysfunction.At present,a lot of research concentrated on how to save in the ischemic semi dark band in brain cells,the opening of the collateral circulation,establishment,reconstruction of blood circulation,blood vessels is newly generated the focal point of research.Endothelial progenitor cells(endothelial progenitor cells EPCs)as adult stem cells can differentiate into mature endothelial cells and is involved in vascular new,breaking the previous that postnatal neovascularization is only by endothelial cell proliferation,migration and collateral Growth and to the traditional view of .In addition,EPCs can accelerate the blood brain barrier(blood-brain barrier,BBB)repair and promote neuronal growth,so it can promote the recovery of neurological function role .Research reported mononuclear cells or CD31 cells homing to ischemic injury and selective integration into the blood capillary,change into mature endothelial cells(ECS).The clinical research in the early stage of this study showed that the prescription of invigorating the spleen and kidney and promoting blood circulation can improve the activity of daily living in patients with cerebral infarction,and the clinical effect is obvious.Therefore,this topic in previous clinical studies based on,SPF male SD rats as the research object,making rat MCAO focal cerebral ischemia reperfusion model,from the neurological score,the percentage of cerebral infarction area of rats than that in rats of peripheral blood EPCs account for foreign week blood mononuclear cells and endothelial cells of rat brain tissue specific marker CD31 antigen protein expression of discussion with strengthening the function of spleen Bushenhuoxue mobilization of EPCs to ischemic zone homing rules,clarify its endogenous neuroprotective mechanism.At the same time,through the discussion of Jianpi Bushen Huoxue Recipe on the index at different time points after cerebral ischemia injury in rats to find the best time window of intervention,provide a scientific basis for clinical medication.Objective:1 Through discussing Jian Pi Bu Shen Huo Xue Recipe mobilization of EPCs to ischemic injury zone homing law,clarify its endogenous protective mechanisms.2 By comparing the neurological function score,cerebral infarction area ratio,peripheral blood EPCs and CD31 protein expression in different time points after I/R to seek the best intervention time window.Method:There were 72 male SD rats in 250-300 g,which were randomly divided into traditional Chinese medicine group,western medicine group,model group,sham operation group,reference line of longa occlusion was used to establish rat cerebral ischemia / perfusion model(MCAO).Each group were divided into ischemia / reperfusion 3h,6h,1D,3D,7d,14 days a total of six observation time points(sham operation group at the same time point observation),6 mice in each group at each time point.before modeling 5D and modeling drug group observation during according to 1ml/100g were given gastric nimodipine suspension liquid and Bushen Jianpi Huoxue Recipe solution.According to longa grade 5 standard score method determination of nerve function Score;TTC staining method was used to detect the cerebral infarction area of rat view;by flow cytometric detection of single nuclear cell suspension was calculated in rats and peripheral blood EPCs for peripheral blood mononuclear cells percentage;the use of immunohistochemical method to detect the rat brain endothelial cell specific markers CD31 antigen protein expression.Result:Rats in each group corresponding nerve function score compared with the sham operation group the score of neural function of rats with different time points were lower than those of the other three groups(P < 0.05).I/R 14d,7d,6h,3h,1D neurological function scores with Rats in each group were lower than I/R 3d(P < 0.05);I/R 14d,3h neurological function scores were lower than I/R 7d,1D,6h,(P < 0.05);I/R 14d neurological function score was lower than I/R 3h(P < 0.05);I/R 6h,3h neural function score were lower than I/R 1D(P < 0.05);I/R 3h neurological function score was lower than I/R 6h(P < 0.05).The nerve function score of the Traditional Chinese medicine group and Nimodipine group was lower than that of the model group(P < 0.05).The neurological function score of the Traditional Chinese medicine group was lower than that of Nimodipine group(P < 0.05).Except the sham operation group,the cerebral infarction area of I/R 3D is larger than that of the I/R 14d,7d,1D,6h,3h(P < 0.05);I/R 14d cerebral infarction area is larger than that of the I/R 7d,1D,6h,3h(P < 0.05);I/R 7d,1D cerebral infarction area is larger than that of the I/R 6h,3h(P < 0.05);I/R 7d cerebral infarction area is larger than that of I/R 1D(P < 0.05).Rats at the same time point compared:The cerebral infarction area in Traditional Chinese medicine group,Nimodipine group,Model group is larger than that of the Sham operation group(P < 0.05);The cerebral infarction area of Model group is larger than that of the Traditional Chinese medicine group,Nimodipine group(P < 0.05);The cerebral infarction area of Nimodipine group is larger than that of the Traditional Chinese medicine group(P < 0.05).The expression of EPCs in peripheral blood showed a tendency to increase,and I/R 14d decreased compared with the former.Before I/R 7d,the expression of EPCs in peripheral blood increased substantially,and the I/R 14d decreased slightly.I/R 14d,3D,1D,6h,3h,EPCs in peripheral blood of all rats in each group were lower than I/R 7d(P < 0.05);I/R 3h,6h,1D in peripheral blood were lower than I/R 3d、14d(P < 0.05);I/R3 d peripheral blood EPCs below I/R 14d(P < 0.05).Compared with the corresponding time points in each group,the expression of EPCs in the peripheral blood of Sham operation group was lower than that in the Model group,Nimodipine group,and the Traditional Chinese medicine group(P < 0.05);In Model group,the expression of EPCs was lower than that of Nimodipine group in the same period,and the Traditional Chinese medicine group(P < 0.05);The expression of EPCs in the peripheral blood of Nimodipine group was lower than that in the same period of the Traditional Chinese medicine group(P < 0.05).Each group at any time point(with the exception of I/R 14d)in brain tissue of rats with CD31 expression showed an increasing trend.Rats in each group I/R 14d,3D,1D,6h and 3H in brain tissue of CD31 were lower than I/R 7d(P < 0.05);I/R 3h,6h,1D in brain tissue of CD31 were lower than I/R 3d and 14 days(P < 0.05);CD31 in brain tissue of I/R 14d was lower than I/R3d(P < 0.05).Compared with the corresponding time points in each group,the brain tissue of CD31 expression in the Sham operation group was lower than that in the Model group,Nimodipine group,and the Traditional Chinese medicine group(P < 0.05);In the Model group,the brain tissue of CD31 expression was lower than that of Nimodipine group in the same period,and the Traditional Chinese medicine group(P < 0.05);The expression of CD31 in Nimodipine group was lower than that in the same period of the Traditional Chinese medicine group(P < 0.05).Conclusion:1 The prescription of strengthening spleen nourishing kidney and activating circulation can obviously improve ischemia/reperfusion rats nerve function score,reduce the area of cerebral infarction;2 The prescription of strengthening spleen nourishing kidney and activating circulation can significantly improve the level of EPCs in peripheral blood of rats with ischemia / reperfusion injury;3 The prescription of strengthening spleen nourishing kidney and activating circulation can significantly enhance the expression of CD31 antigen in brain tissue of rats with ischemia / reperfusion injury;4 The mechanism of improve ischemia/reperfusion rats nerve function score,reduce the area of cerebral infarction may be related to Promote angiogenesis in brain tissue and the establishment of collateral circulation by Increasing the level of EPCs in peripheral blood and enhancing the expression of CD31 antigen in brain tissue. |
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