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补肾强肝活血法治疗缺血性脑卒中的实验研究
Experimental Study on the Theory of Tonifying Liver and Kidney and Activating Blood Circulation for Cerebral Ischemic Stroke
【作者】 刘骏;
【导师】 张发荣;
【作者基本信息】 成都中医药大学 , 中医内科学, 2009, 博士
【摘要】 目的:通过建立缺血性中风后动物模型,探讨补肾强肝活血法思想指导下的回春偏瘫方对缺血性中风后神经再生作用的影响,为其修复缺血性脑卒中后的神经功能提供实验依据。方法:用线栓法造成大脑中动脉阻塞(middle cerebral artery occlusionMCAO)再灌注模型,造模成功后随机分为假手术组,模型组、回春偏瘫方高、中、低剂量(回高组/回中组/回低组)、西药尼莫地平组(西药组),连续给药14天,治疗结束后观察大鼠一般情况和体重改变,按照改良Berdson评分分级法观察神经行为学的改变,TTC染色观察大鼠脑梗死体积变化和脑细胞形态学改变,开颅取脑制成10%脑匀浆溶液,测定脑内SOD、MDA、Glu的水平,用酶联免疫吸附法测定缺血再灌注大鼠颅内NGF水平,免疫组化染色观察脑源性神经营养因子(BDNF),生长相关蛋白-43(GAP-43),突触素p38(SYN)在颅内的表达。结果:经过MCAO再灌注造模后,大鼠均出现体重下降,神经功能缺失评分明显升高(p<0.01);实验结束后,与模型组比较,治疗各组体重有所恢复,神经功能缺失情况得到不同程度改善,各治疗组脑梗死体积和脑梗死体积百分比均有不同程度的减少,有极显著的差异(P<0.01):各治疗组脑组织中Glu水平均有所降低,有明显差异性(P<0.05),中剂量组或高剂量组脑内SOD水平明显升高(P<0.01),MDA水平明显降低(P<0.01),但治疗低剂量组仅对SOD的升高有明显效果(P<0.05),但对MDA的改变效果不明显(P>0.05);治疗各组脑内NGF水平均高于模型组(P<0.01),尤其是中剂量组效果最佳,疗效与西药组相当(P>0.05);治疗各组与单纯模型组病理切片结果相比有不同程度改善,其中以中剂量治疗组改善略明显;缺血性卒中大鼠脑内BDNF表达显著增强(P<0.01),其中尤以中剂量组疗效最佳,与西药组无明显差异性(P>0.05);各剂量组脑内GAP-43表达增强(P<0.01),其中高剂量和中剂量组表达水平与西药组相当,无明显差异性(P>0.05);但就脑内SYN表达而言,仅中剂量组和高剂量组脑内SYN表达显著增强(P<0.05),与西药组相当(P>0.05)。结论:回春偏瘫方作为体现补肾强肝活血法的有效方剂,具有促进MCAO再灌注后大鼠神经功能恢复的作用。其作用机理与减小大鼠脑梗死体积,降低脑内Glu、MDA水平,升高脑内SOD、NGF水平,有效改善脑病理形态学,提高颅内BDNF、GAP-43和SYN的表达。
【Abstract】 Objective:To investigate the effects of Huichunpiantan Decoction(HD) under guidance of the theory of tonifying liver and kidney and activating blood circulation on treating stroke models of rats after experimental cerebral ischemia (CI).Methods:The rats after successful modeling the models of middle cerebral artery occlusion (MCAO) were randomized into six groups:sham operation group, model group, high dose group, middle dose group, low dose group and control group. After 14 days treatment course, the changes of weight, behavior, infarct size, the structures of brain tissue, the contents of SOD, MDA, GIu, NGF in brain, and the expressions of BDNF, GAP-43, SYN were investigated.Results:①HD decreases neural functional deficits in the evaluation of rats and the score with long medication was lower, suggesting that medication can even better improve neural functional deficits;②HD could obviously reduce the infarct size;③HD could increase SOD activity, reducing MDA and Glu content in brain;④HD could markedly increase the content of NGF in brain;⑤HD could protect the structures of brain tissue in experimental cerebral ischemia rats;⑥HD could significantly promote the expression of BDNF in brain;⑦HD could promote the expressions of Gap-43 and SYN.Conclusions:The experiment shows that HD as an effective decoction by tonifying liver and kidney and activating blood circulation can improve neuronal functional recovery in CI.