| Stroke is a kind of nervous system injury disease which has high morbidity, mortality, disability rate and recurrence rate. According to the World Health Organization(WHO) survey results show that mortality of stroke is the third after cancer and myocardial infarction. The latest statistics show that there are 1.5 million new cases and 1 million death case of stroke in China every year, disability rate is as high as 75% among the 6million live patients after the illness, which brings the heavy burden to family and society. Most stroke is ischemic stroke in Chinese population. In the development of mature mammalian nervous system, the discovery of neural stem cells brings the hope of repairing central nervous system(CNS)injury. Therefore, according to a variety of drugs or other ways, nerve functional recovery could be recoveried by means of activation of endogenous NSCs, enhancing or adjustment the stem cell regeneration response, promoting differentiation and migration to the target area. The endogenous NSCs mobilization treatment will be the future direction of ischemic cerebral stroke. Kangnaoye consists of astragalus membranecues,salvia miltiorrhiza, puerarin, uncaria, rhizoma ligustici wallichii, notoginseng powder and so on, studies have shown that Kangnaoye Recipe could promote the angiogenesis and antioxidant. This experiment is to observe the effect of Kangnaoye Recipe on the expression of nerve regeneration specific marker Brdu, GFAP, Nestin and Nestin mRNA expression by making cerebral ischemia reperfusion model, so as to do further research in the aspect of Kangnaoye Recipe promoting nerve regeneration after cerebral ischemia reperfusion, provide powerful basis to clinical treatment as soon as possible.The 150 rats were randomly divided into sham-operated group, model group, high-dose Kangnaoye group, middle-dose Kangnaoye group and low-dose Kangnaoye group(24,12,6 g.kg-1.d-1). The modle of cerebral ischemia reperfusion(I/R) was induced by middle cerebral artery occlu sion(MCAO) and recanalization. Neurological symptom score were evaluated in 2 h and 24 h after ischemia. HE staining of the cells was used to observe the morphology changes in hippocampus CA1 area. TTC staining in 1d after ischemia was to observe the volume change of cerebral infarction in rats. Glial fibrillary acidic protein(GFAP) protein and Nestin protein were observed by immunohistochemical method in CA1 area.Immunofluorescence assays were applied for detecting Brdu/nestin contents of necrotic area in each time point and the expression of Nestin mRNA were used to be measured by in situ hybridization.Compared with shamed group, the symptom of neurological defe ction, the volume of cerebral infarction and the interstitial edema aro und neurons were very obviously after the reperfusion in rats of cere bral ischemia in model group, the number of cells around hippocamp al CA1 area decreased, with scattered arrange, broadening the gap be tween cells, thick dying nucleus, nerve function defect score of posto perative rats were higher(P < 0.01); compared with the model group,it could be noted in Kangnaoye recipe preconditioning groups, the v olume of cerebral infarction and pathological morphological changes were decreased(P < 0.05 or P < 0.01). The groups pretreated with Kangnaoye had a lower expression of GFAP around hippocampal CA1 area, at the same time the expression of Nestin, Nestin mRNA and Brdu expression were higher, which has significant differences(P <mRNA expression at the same time. Therefore, enhancing cell proliferation as well as promoting the neural stem cells regeneration and differentiation may be the way to promote the nerve regeneration for Kangnaoye recipe. |
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