| AIM: Intracerebral hemorrhage (ICH ) is dangerous category in acute cerebrovascular disease,with higher mortality rate.About 60-80% cases left over nerve dysfunction certainly degree, even though fortunately survived. Nerve dysfunction caused by ICH is because of degeneration necrosis and apoptosis of neuronal around hematoma. Neurotrophic factor(NTF) have many physiological function that accelerate neuronal survival and regulate synapse plasticit and stimulate axon growth.Neural stem cells(NSC)with multidifferentiation potency are seed cells that nerves regenerate.After brain injury,neuron. astroglia. oligodendrocyte that NSC proliferate and differentiate into could restore structure and integrate function.Althoug ICH could stimulate neural protection of CNS by accelerating release of endogenous NTF and activation of endogenous NSC,however NTF perhaps release transiently and activation NSC differentiate into mass neuroglias and a few neurons . Therefore ,to promote the expression and secretion of NTF ,and alterate differentiated direction of NSC would become action target of various drug treating ICH. Nao yi-an(NYA) is a kind of traditional Chinese medicine treating ICH (acute stage),and had valid clinical effect.On the basis of previou investigation,this experiment want to demonstrate the effects of NYA on secretion of endogenous brain-derived neurotrophic factor(BDNF) and glial cell line-derived neurotrophic factor(GDNF) and on NSC differentiation .. hypoxia impairment protection, so as to investigate the brain protective function of NYA on ICH.METHODS:ln vivo (l)The intrscerebral hemorrhage(ICH) animal models in rat have been established by injecting 0.4U type VII collagenase into the right globus pallidus (GP) by use of stereotaxic apparatus. (2)The distribution and expression difference of BDNF^GDNF in the ICH rat brains and effect of NaoYiAn(NYA) have been investigated by immunohistochemistry technique.(3) The distribution and expression difference of BDNF. GDNF in the ICH rat brains and effect of NaoYiAn(NYA) have been investigated by in situ hybridizationtechnique.In vitro (1) The cultured neural stem cells(NSC) have been taken from the newborn Sprague-Dawley(SD) rats at postborn 3-5 days.(2) Preparation of NYA serumrserum have been separated from blood of rat that have been ingested with NYA after three days.(3) Hypoxia NSC models:the NSC have been cultured for six hours at anaerobic incubator(37癪 95%N2> 5%CO2).(4) To study the effection of NYA on differentiation and existence of NSC in vitro by immunocytochemistry and MTT technique.(5) Using immunocytochemistry to study the expression of BDNF and GDNF in normal NSC and NSC that had been damaged by hypoxia.RESULTS: (1) No positive BDNFand GDNF cells presented in brain of normal controK sham-operated rats.Both BDNF and GDNF positive cells were to limited to hematom and prihematom regions after ICH, and had nothing in contralateral region of injury N cortex > hippocampus. Both BDNF and GDNF protein begun express at 2h, reached the peak at Id, and decrease at 4d, expressed weakly at 7d after ICH injury, but positive GDNF cells disappeared at 14d after ICH injury, and at the same time, positive BDNF cells were not different from that at Td.BDNF were mainly expressed in activated microglia around prehematom by BDNFand OX-42 immuno-fluorescence-dual-labelling method ,and From morphology ,GDNF were mainly expressed in actived astroglia around prehematom. The BDNF and GDNF protein expression level of NYA group were higher than model group at ld^ 4d after ICH injury(P<0.05).To sum up, NYA could up-regulated the expression of BDNF and GDNF protein. (2) Expression of BDNFmRNA and GDNFmRNA were discovered in cortex N hippocampus > striatum of normal controls> sham-operated rats, but expression of GDNFmRNA were very weak.after 2h of ICH,expression of BDNFmRNA and GDNFmRNA increased in cortex -. hippocampus ? striatum -. basis forebrah^ substantia nig^a^ corpus callosum> perihematom ,but no expression signals were found in the hematoma region, and afte...
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