脑心通对局灶性脑缺血大鼠行为学和脑组织肿瘤坏死因子α的影响

Effect of naoxintong on the behavioral changes and tumor necrosis factor-alfa in rats with focal cerebral ischemia

目的:探讨脑心通对大鼠局灶性脑缺血损伤后行为学及脑组织肿瘤坏死因子α表达的影响。方法:实验于2004-03/11在河北医科大学第二医院神经内科实验室进行。取成年健康雄性SD大鼠150只,制造大鼠局灶性大脑中动脉阻断造模手术后,将大鼠随机分为5组,即脑心通4g/kg组、2g/kg组、1g/kg组、生理盐水和假手术组,各组又分为6,24,48,72h,7d5个亚组,每亚组均为6只大鼠。脑心通各剂量组均为灌胃给药,生理盐水组给同体积的生理盐水,于相应时间点进行行为学评分后处死动物快速取脑,观察脑含水量的变化,病理切片苏木精-伊红染色观察组织病理学改变,免疫组织化学的方法测定肿瘤坏死因子α的动态变化。结果:实验中有8只大鼠死亡,后随机补充,进入结果分析仍为150只大鼠。①行为学评分:大鼠在大脑中动脉阻断后手术对侧肢体存在不同程度瘫痪,在术后48和72h时神经功能缺损最为明显,7d时神经功能基本恢复正常。②病理观察:除假手术组外,其余各组在脑梗死后6h局部可见少量散在炎性细胞浸润;梗死后48h,炎性细胞浸润明显增多;并持续到7d。③脑组织含水量:与假手术组相比,其他各组在各时间段均增加。术后24,48h脑心通4g/kg组为(78.95±1.36)%,(79.44±1.01)%,脑心通2g/kg组为(79.03±0.87)%,(80.46±1.54)%,低于同期的脑心通1g/kg组和生理盐水组[(79.21±0.68)%,(82.02±1.76)%,(79.30±1.11)%,(82.68±0.91)%,P<0.05]。④肿瘤坏死因子α阳性细胞表达:与假手术组相比,其他各组在各时间段均增加,梗死后6h开始增多,48h达高峰。术后24,48h脑心通4g/kg组为(27.6±2.1),(18.4±1.5)个,脑心通2g/kg组为(38.4±2.1),(23.2±3.3)个,低于同期的脑心通1g/kg组和生理盐水组[(50.4±5.6),(30.8±5.3),(52.8±3.7),(33.0±4.7)个,P<0.05]。结论:脑心通可能通过减轻脑水肿,降低脑内肿瘤坏死因子α的表达减少炎性细胞的浸润,从而对大鼠缺血脑组织损伤产生保护作用。

AIM:To study the effect of naoxintong on the behavioral changes and the expression of tumor necrosis factoralfa in rats with focal cerebral ischemia.METHODS:The experiment was performed in the Department of Neurology,Second Hospital of Hebei Medical University between March and November 2004.A total of 150 SD male rats were selected to establish focal cerebral ischemia models by middle cerebral artery occlusion.Rats were divided randomly into five groups:sham operation group,saline group,4 g/kg naoxintong group,2 g/kg naoxintong group and 1 g/kg naoxintong group.Rats in each group were divided into groups of 6,24,48,72 hours and 7 day after middle cerebral artery occlusion,with 6 rats in each group.Rats in naoxintong and saline groups were injected intragastrically with the same dose of naoxintong and saline,respectively.The rats were killed to extract brain tissue following praxiology scoring;the changes of brain water content were measured;the changes of pathology were observed with hematoxylin and eosin staining method;immunohistochemistry method was used to detect the dynamic changes of tumor necrosis factoralfa. RESULTS:Eight rats died in the experiment,and another eight were selected;finally,a total of 150 rats were involved in the result analysis.①The score on praxiology:Palsy in different degree existed in limbs after middle cerebral artery occlusion.The neurologic impairment reached to the top at hours 48 and 72,trended to the normal at hour 7.②Pathological changes:a small quantity of inflammatory cellular infiltration in local parts indicated in each group except sham operation group at hour 6 following cerebral infarction.After 48 hours,the inflammatory cellular infiltration increased until day 7.③Water content in brain increased in each group except sham operation group.The water content in brain was(78.95±1.36)%and(79.44±1.01)%at hours 24 and 48 in 4 g/kg naoxintong group,(79.03±0.87)%and (80.46±1.54)%in 2 g/kg naoxintong group,(79.21±0.68)%and(82.02±1.76)%in 1 g/kg naoxintong group,and (79.30±1.11)%and(82.68±0.91)%in saline group(P< 0.05).④The expression of tumor necrosis factoralfa increased in each group except sham operation group,which increased at hour 6 after cerebral infarction and reached to the peak at hour 48.The expression of tumor necrosis factoralfa was 27.6±2.1 and 18.4±1.5 in 4 g/kg naoxintong group,38.4±2.1 and 23.2±3.3 in 2 g/kg naoxintong group,which was lower as compared with that in 1 g/kg naoxintong group(50.4±5.6,30.8±5.3) and saline group(52.8±3.7,33.0±4.7) at the same time points(P< 0.05).CONCLUSION:Naoxintong may protect brain tissue by alleviating cerebral edema, reducing the expression of umor necrosis factoralfa and the inflammatory infiltration of cells in rats with focal cerebral ischemia.