【作者】 杜可；

【导师】 蔡光先；

【作者基本信息】 湖南中医药大学 ， 中医内科学， 2010， 博士

【摘要】 目的：探讨超微脑得健对脑缺血大鼠脑内神经再生相关信号GAP-43、Rho的影响,揭示其可能分子机制。方法：1.通过古代和现代文献整理,复习缺血性脑血管疾病的中西医认识和防治进展。学习经典补阳还五汤的现代药理作用。初步认识一些调节中枢神经再生的信号分子。2.采用MCAO法复制大鼠局灶性脑缺血模型,将140只成年SD雄性大鼠随机分为空白对照组(即正常组)、假手术组、模型组、补阳还五汤组、脑得健原方组,超微脑得健全量组,超微脑得健1/3剂量组。分别给予不同处理,在1d、3d、7d、14d不同时间点处死动物,TTC染色法观察大鼠脑梗死情况并进行神经功能评分,采用免疫组化、RT-PCR法检测超微脑得健对GAP-43、Rho蛋白表达的影响。结果：1.文献研究缺血性脑血管疾病是中老年常见疾病。1.1中医常见气虚血瘀型,气虚是本,血瘀为标；常用益气活血、祛瘀生新法治疗。1.2西医多见血栓型,多以物质堵塞腔管,血液流变学障碍为表现。现代西医认为促进中枢神经再生为脑缺血后主要治疗目标。多种内外科治疗效果不佳或存在明显使用缺陷。1.3补阳还五汤具多种现代药理学作用从而促进脑缺血后神经功能恢复。1.4 GAP-43是目前国际公认的促中枢神经元再生的标记性蛋白,小G蛋白Rho则为新近的抑制中枢神经再生分子信号之一2实验研究2.1.TTC法检测造模成功的大鼠脑组织梗死面积大于健侧,呈苍白梗死灶。2.2脑得健干预的各实验组脑组织病理切片呈好转状态。2.3给药3天后,脑得健干预组各时间点神经功能积分均显著低于模型组(P<0.05),超微脑得健1/3剂量组各时间点神经功能积分和补阳还五汤及脑得健各组比(P>0.05)。2.4免疫组化结果示各治疗组GAP-43表达水平在各时间点均高于对照组,两者之间差异有统计学意义(P<0.05)2.5给药3d后各治疗组动物脑组织中GAP-43 mRNA含量较同时段模型组比较明显增加(p<0.05),脑得健方组和补阳还五汤组比较差异无统计学意义(p>0.05),各药物组比较差异无统计学意义(p>0.05)。2.6给药3d后各治疗组动物脑组织中Rho mRNA含量较同时段模型组比较明显降低(p<0.05),脑得健方各组和补阳还五汤组比较差异无统计学意义(p>0.05)。结论:1脑得健方能改善大鼠局灶性脑缺血神经功能缺失症状。2脑得健能促进MCAO大鼠脑缺血损伤后GAP43蛋白表达同时抑制Rho蛋白表达。可能是其促脑缺血后神经再生和功能恢复的治疗机制之一。3超微脑得健1/3剂量和传统脑得健饮片以及传统补阳还五汤作用相当。更多还原

【Abstract】 Objectives:Study on the effect of UNDJ on neural regeneration signaling molecule GAP-43、Rho during cerebral infarction convalescence to provide the possible anti-ischemic mechanism.Methods:1. Through the collection of ancient and modern literature, review the process of prevention and treatment on ICVD both in Western and Chinese medicine. Study the modern pharmacological action of classical decoction Buyang Huanwu. Have a preliminary grasp of some signaling molecules on neural regeneration.2. Rat focal cerebral ischemia was induced by MCAO, After Rats model of focal cerebral ischemia induced, All adult male SD rats were randomly divided into 7 groups including naive, shame-operated, ischemic, ischemic with BYHW, ischemic with NDJ, ischemic with UNDJ and ischemic with 1/3UNDJ groups. Each group have 20 rats. ALL the rats were given different treatment and were killed in 1,3,7or14day after operation. To observe the infarct size by TTC and evaluation the nerve function, the expressions of GAP-43、Rho were measured by immunohistochemical staining both with RT-PCR.Results:1. Literature Study:ICVD is a common complaint among the elderly.1.1 Pattern of qi deficiency and blood stasis is often seen in traditional Chinese medical science, qi deficiency is etiological factor while blood stasis is symptomatic factor. We replenishing qi and activating blood in order to dispelling stasis to promote regeneration.1.2 Cerebral thrombosis is often seen in western medicine, Obstruction of a blood vessel and hemorheology by an embolus usually can be seen. Modern medicine shows that the main therapeutic goal during cerebral infarction convalescence is to help the centrum neural regeneration.But now all the treatment effects are have deficient and below the mark.1.3 BYHW have many modern pharmacological action to improve the neurological deficit.1.4 GAP-43 is a internationally recognized neural regeneration signaling protein,and small G-protein Rho is one of the latest rejection capability neural regeneration signaling molecule. 2. Experimental Study:2.1 After ischemia, the pale infarction in hemorrhage brain can be seem greater than the brain offside by TTC detection。2.2 All the pathological sections with NDJ treatment groups show a improved tendency.2.3 All the NDJ treatment groups can significant improve neurological deficits at 3,7,14days (P< 0.05), the 1/3UNDJ group,BYHW group and decoction NDJ group have no obvious differences at each time point (P>0.05)2.4 Immunohistochemical staining showed that the GAP-43 expressions level in all treatment groups is higher than the ischemic groups at each time point, there was a significant difference(P<0.05).2.5 RT-PCR showed that the GAP-43 mRNA level in all treatment groups is higher than the ischemic groups after 3d time point, there was a significant difference(P<0.05). the 1/3UNDJ group,BYHW group and decoction NDJ group have no obvious differences at each time point (P>0.05)2.6 RT-PCR showed that the Rho mRNA level in all treatment groups is lower than the ischemic groups after 3d time point, there was a significant difference(P<0.05). the 1/3UNDJ group,BYHW group and decoction NDJ group have no obvious differences at each time point (P>0.05) Conclusions:1. Decoction NDJ can improve the neurological deficits.2. NDJ can promote GAP43 expression while resistant Rho expression after cerebral ischemia of MCAO rats,which maybe one of anti-ischemic mechanism of the drug.3.1/3 UNDJ is affected with a considerable amount of the role of traditional pieces.更多还原