回医扎里奴思方对痰热腑实证脑缺血再灌注大鼠肿瘤坏死因子-α和内皮素-1表达的影响

Effect of Hui medicine Zhali Nusi Fang on expression of TNF-α and ET-1 in the heat-phlegm and sthenic-fu syndrome rats with cerebral ischemia-reperfusion

目的探讨扎里奴思方对痰热腑实证脑缺血再灌注大鼠脑内肿瘤坏死因子(TNF)-α和内皮素(ET)-1表达的影响。方法将60只雄性SD大鼠随机分为假手术组、模型组、尼莫地平组、扎里奴思方组,每组15只。除假手术组外,其余各组均给予高脂喂养4 w后,再给予10%的自体粪便1 ml/100 g灌胃,1次/d,连续3 d,造成痰热腑实证模型;再采用线栓法制备大鼠大脑中动脉阻塞(MCAO)。大鼠给药剂量为尼莫地平组10.8 mg/kg、扎里奴思方组1.54 g/0.1 kg,制备MCAO模型前3 d灌胃给药。大鼠分别于缺血再灌注后1、3、7 d取脑;取材前进行脑组织含水量测定,采用免疫组化法检测脑缺血侧海马区TNF-α、ET-1表达,RT-PCR检测TNF-α mRNA和ET-1 mRNA的表达。结果与假手术组比较,模型组脑组织含水量增高(P<0.01)、TNF-α、ET-1阳性细胞和TNF-α mRNA、ET-1 mRNA表达明显增高(P<0.01);与模型组比较,尼莫地平组和扎里奴思方组脑组织含水量降低、TNF-α、ET-1阳性细胞数减少,TNF-α mRNA、ET-1 mRNA表达降低(P<0.01)。与尼莫地平组比较,扎里奴思方7 d组脑组织含水量降低、TNF-α、ET-1阳性细胞数减少和TNF-α mRNA、ET-1 mRNA表达降低(P<0.05)。结论扎里奴思方对痰热腑实证脑缺血再灌注大鼠脑神经元起保护作用,其机制可能与抑制大鼠脑内TNF-α和ET-1的表达有关。

Objective To investigate the effect of Hui medicine Zhali Nusi Fang on expression of TNF-α and ET-1 in the heat- phlegm and sthenie-fu syndrome rats with cerebral ischemia-reperfusion. Methods 60 male SD rats were randomly divided into： sham, mod- el, nimodipine and Zhalinusi Fang groups （n = 15 ）. Except sham group, the other groups were fed with high fat-diet for 4 weeks. Then, rat stools were administered intragastrically once a day for three days to make the heat-phlegm and sthenic-fu syndrome, then focal cerebral is- chemia models were established by middle cerebral artery occlusion with nylon thread. Rats were given with nimodipine （10.8 mg/kg） , Zhali Nusi Fang （ 1.54 g/0. 1 kg） by i.g. before the model preparation. Brain were taken at 1, 3, 7 days respectively to determine the brain water content, and then the expression of TNF-ct and ET-I was detected by immunohistochemistry and RT-PCR. Results Compared with sham group, brain water content of model group rats were increased（ P〈0.01 ） , TNF-α and ET-1 positive cells and the expressions of TNF-α and ET-1 mRNA were up-regulated （P〈0.01）. Compared with model group, brain water content of Zhali Nusi Fang and nimodipine groups were reduced, TNF-α and ET-1 positive cells and the expressions of TNF-α and ET-1 mRNA were decreased （P〈0.01）. Compared with ni- modipine group, TNF-α and ET-1 positive cells and the expressions of TNF-ot and ET-1 mRNA of Zhali Nusi Fang 7 d group were decreased （P〈0.05）. Conclusions Zhali Nusi Fang has protective effects against the heat-phlegm and sthenic-fu syndrome rats after cerebral ische- mia-reperfusion, which may be related with inhibiting the expression of TNF-α and ET-1.