中风胶囊对脑出血大鼠神经功能、脑组织细胞凋亡及肿瘤坏死因子-α的影响

Experimental research of Zhongfeng capsule on neurological function,apoptosis and TNF-α in brain tissue of rat with intracerebral hemorrhage

目的探讨中风胶囊对脑出血大鼠神经功能、脑组织细胞凋亡及肿瘤坏死因子-α(TNF-α)的影响。方法选取SD大鼠104只,随机分为5组,即正常组8只,模型对照组、假手术组、中风胶囊大剂量组、中风胶囊小剂量组各24只,后4组按手术后处死时间随机分别分为1、37、d组,每组8只。采用自体血注射法制作大鼠实验性脑出血模型。模型对照组在造模后自由饮食;中风胶囊大、小剂量组在造模后分别给予中风胶囊2.7、1.8 g/(kg.d)灌胃;假手术组操作同模型对照组,但只进针,不注射自体血,术后自由饮食;正常组自由饮食,不做特殊处理。各组分别于第1 d(24 h)、第3 d(72 h)和第7 d进行神经功能评分,随后处死取出脑组织标本,分别采用末端脱氧核糖核酸转移酶介导的缺口末端标记法(TUNEL法)和免疫组化链霉菌抗生物素蛋白-过氧化物酶(SP)法测定血肿周围细胞凋亡和血肿周围TNF-α蛋白表达。结果各组第1、3、7 d神经功能评分、TUNEL阳性细胞比例及TNF-α阳性细胞比例均低于模型对照组(P<0.01);中风胶囊大剂量组第37、d神经功能评分、TUNEL阳性细胞比例及TNF-α阳性细胞比例均低于中风胶囊小剂量组(P<0.05),中风胶囊大剂量组第1 d TUNEL阳性细胞比例低于中风胶囊小剂量组(P<0.01,P<0.05);中风胶囊大剂量组第7 d神经功能评分、TUNEL阳性细胞比例及TNF-α阳性细胞比例均低于本组第1d(P<0.01,P<0.05)。结论中风胶囊能促进脑出血急性期大鼠神经功能恢复,降低脑组织中TNF-α蛋白的表达,从而抑制细胞凋亡,具有减轻炎症及脑组织损伤的作用。

Objective To investigate the effect of Zhongfeng capsule on neurological function,apoptosis and TNF-α in brain tissue of rat with intracerebral hemorrhage.Methods 104 SD rats were randomly divided into five groups of normal group（NG）,sham operated group（SO）,model group（MG）,high-dose drug group（HG） and low-dose drug group（LG）.8 rats were in NG,24 in other groups.Experimental rat models of intracerebral hemorrhage are made by autoblood injection method.After establishment of model,rats in HG and LG were given 2.7 g/（kg·d） and 1.8 g/（kg·d） Zhongfeng capsule.Neurological function score was carried on at 1 d,3 d and 7 d after operation,then brain tissue was removed.Apoptosis degree of haematoma peripheral cells was observed with tagged deoxynucleotide transferase mediated dUTP nick end labeling（TUNEL）.The proteins of TNF-α expression were detected with immunohistochemical method.Results Neurological function score,TUNEL positive cell ratio and TNF-α positive cell ratio in each group were lower than those in NG.Neurological function score,TUNEL positive cell ratio and TNF-α positive cell ratio in HG were lower than those in LG at 3 d and 7 d after operation（P0.01）.TUNEL positive cell ratio in HG was lower than that in LG at 1 d after operation（P0.05）.The indexes above mentioned at 7 d were lower than those at 1 d after operation in HG（P0.05）.Conclusion Zhongfeng capsule can promote neural functional recovery and effectively decrease TNF-α protein expression in acute stage of intracerebral hemorrhage,inhibit apoptosis and lighten inflammatory reaction,so as to relieve the brain tissue damage.