【摘要】 目的:观察碱性成纤维细胞生长因子及其受体在体外模拟脑缺血再灌注损伤中的表达变化及以天麻和钩藤为主的中药制剂——抗呆Ⅰ号对其的影响。方法:实验于2002-03/2004-04在科学实验中心完成。先分别进行大鼠大脑皮质星形胶质细胞和神经元的分离纯化培养及体外模拟脑缺血再灌注损伤模型的建立,然后分别进行下列实验:①按随机数字表法将传代后培养5d的星形胶质细胞分为正常对照组、缺血再灌注模型组和缺血用药组(应用抗呆Ⅰ号),在体外模拟脑缺血4h和再灌注3h,18h,24h,36h,48h和72h后行碱性成纤维细胞生长因子的免疫细胞化学染色。②用再灌注18h后收集的星形胶质细胞条件培养液、经抗呆Ⅰ号作用的星形胶质细胞条件培养液及星形胶质细胞条件培养液与抗呆Ⅰ号联合应用以1∶5的浓度来培养损伤后的神经元,再对其培养的神经元行碱性成纤维细胞生长因子受体的免疫组化染色。结果:①体外培养大鼠的大脑皮质星形胶质细胞在模拟脑缺血再灌注损伤后各时相点分泌碱性成纤维细胞生长因子的能力均显著高于正常对照组(P<0.05～0.01);除再灌注72h外,其余各时相点缺血用药组碱性成纤维细胞生长因子的表达水平均显著高于缺血再灌注模型组(P<0.05～0.01)。②星形胶质细胞条件培养液组、经抗呆Ⅰ号作用的星形胶质细胞条件培养液组和联合应用组脑缺血再灌注损伤后各时相点的碱性成纤维细胞生长因子受体表达水平大多显著高于缺血再灌注模型组(P<0.05～0.01),均在再灌注18h时达到高峰,其作用强度为经抗呆Ⅰ号作用的星形胶质细胞条件培养液>联合应用>星形胶质细胞条件培养液。结论:体外模拟脑缺血再灌注损伤使星形胶质细胞表达碱性成纤维细胞生长因子的能力增强,星形胶质细胞条件培养液能促进受损神经元碱性成纤维细胞生长因子受体的表达。抗呆Ⅰ号可增强受损神经组织的分泌功能,促进其损伤后的修复。更多还原

【Abstract】 AIM: To observe expression of basic fibroblast growth factor (bFGF) and its receptor in establishing cerebral ischemia-reperfusion models in vitro and the influence of Chinese herb compounds of gastrodia rhizome and gambir plant (named Kangdai Ⅰ). METHODS: The experiment was performed at Scientific Experimental Center from March 2002 to April 2004. Firstly, astrocyte and neurons of cerebral cortex of rats were isolated, purified and cultured; Cerebral ischemic reperfusion injury models were established in vitro, and then the followed trials were conducted, respectively: ①Astrocytes cultured for 5 days after passage were randomly assigned into normal control group, ischemia-reperfusion model group and ischemia medication group (Kangdai Ⅰ). The bFGF was determined with immunocytochemical stain after modeling cerebral ischemia for 4 hours and reperfusion for 3, 18, 24, 36, 48 and 72 hours in vitro. ②Injured neurons were cultured with astrocyte conditioned medium collected after 18-hour reperfusion, astrocyte conditioned medium affected by Kangdai Ⅰ and compound of astrocyte conditioned medium and Kangdai Ⅰ with the 1∶5 concentration, and then the bFGF receptors of cultured neurons were determined with immunocytochemical stain. RESULTS: ①Ability of astrocytes of cerebral cortex of rats cultured in vitro in secreting bFGF after modeling cerebral ischemia-reperfusion injury at each time phase was significantly higher than that in the normal control group (P < 0.05-0.01). Except reperfusion for 72 hours, expressive level of bFGF in ischemia medication group was markedly higher than that in the ischemia-reperfusion model group (P < 0.05-0.01). ②Expressive levels of bFGF receptors at each time phase after cerebral ischemia-reperfusion injury in the astrocyte conditioned medium group, astrocyte conditioned medium affected by Kangdai Ⅰ group and compound group were remarkably higher than that in the ischemia-reperfusion model group(P < 0.05-0.01). It reached the peak at hour 18 after reperfusion. The effect intensity from high to low was astrocyte conditioned medium affected by Kangdai Ⅰ, compound and astrocyte conditioned medium. CONCLUSION: The bFGF expression of astrocytes increases in the damage induced by simulating cerebral ischemia and reperfusion in vitro. Astrocyte conditioned medium can accelerate the expression of bFGF receptors of injured neurons. Kangdai Ⅰ can enhance secretary function of injured nervous tissue and accelerate post-trauma reparation.更多还原