The Approach About The Action Of Naodesheng Compound Recipe And It's Effective Constituent's Mechanism

OBJECTIVE This study is designed to check up Naodesheng,Sanqi and Chuanxiong's protective effect on cerebral ischemia in MCAO rats,and then compare with each other.Hydroxysafflor yellow A is one of the main active ingredients extracted from Honghua which is a constituent of Naodesheng.I design to investigate the primary molecular base underlying Hydroxysafflor Yellow A on angiogenesis of brain tissue in rats with focal cerebral ischemia with the method of Real-time PCR.METHODS The establishment of focal cerebral ischemia in rat was followed by the description of Zea Longa,et al.Healthy adult male Sprague-Dawley rats, weighting from 260g to 280g,were randomly divided into nine groups as follows:the sham-operated group(n=10),the model group(n=10),Naodesheng group(n=9), Naodesheng crude extract group(n=10),Naodesheng effective constituent group(n=10),Sanqi crude extract group(n=10),Sanqi effective constituent group(n=10),Chuanxiong crude extract group(n=10),Chuanxiong effective constituent group(n=10).rats are administered by mouth immediately after the onset of MCA occlusion and they are administered five times at the interval of every 30 minutes.At 24 hour post MCA occlusion,the behavior deficits were observed,and then infarction was stained by TTC.The establishment of focal cerebral ischemia in rat was followed by the description of Zea Longa,et al.Healthy adult male Sprague-Dawley rats,weighting from 260g to 280g,were randomly divided into three groups as follows:the sham-operated group(n=3),the model group(n=3) and HSYA group(n=3).12mg/kg of HSYA was injected via sublingular vein immediately after the onset of MCA occlusion and followed by the presence of HSYA at the dose of 12 mg/kg at the interval of every 30 minutes,respectively,adding up to 60 mg/kg dosage.At 3, 24-hour time point post MCA occlusion,Real time RT-PCR was used to detect mRNA levels of VEGF and HIF-1αgenes.RESULTS 1.The rats in the model group showed significant neurobehavioral obstacles and ischemic brain infarction at 24 hour point after the onset of MCA occlusion compared with those in the sham-operated groups.(P＜0.01)2.Naodesheng as well as Naodesheng crude extract and Naodesheng effective constituent significantly improved ischemia-induced behavioral impairment and reduced the infarct volume(P＜0.01).3.Chuanxiong crude extract and Chuanxiong effective constituent significantly improved ischemia-induced behavioral impairment and reduced the infarct volume (P＜0.05).4.Sanqi crude extract can improved ischemia-induced behavioral impairment(P＜0.05),but it can not reduced the infarct volume.Sanqi effective constituent significantly improved ischemia-induced behavioral impairment and reduced the infarct volume(P＜0.05).5.The validation of microarray implicated that down regulated expression of aFGF,HIF-1α,Flt,bFGF gene in the cortex penumbra of rats with MCAO was significantly reversed by administration of HSYA at immediate early stage(3h) post cerebral ischemia.Similarly,HSYA promoted mRNA expression levels of bFGF and VEGF at 24h point(P＜0.05).Naodesheng,as well as Naodesheng crude extract and Naodesheng effective constituent have good protective effect on cerebral ischemia,and it's effect better than Sanqi and Chuanxiong.CONCLUSION 1.Naodesheng,Sanqi and Chuanxiong all have good effect protective effect on cerebral ischemia.2.molecular basis of angiogenic effect afforded by the presence of HSYA in vivo study,might be related with the beneficial regulation of the transcriptional levels of HIF-1αand VEGF,positive growth factors on angiogenesis in rats cortical penumbra.