化瘀通络汤对局灶性脑缺血-再灌注损伤大鼠的脑神经保护作用

Neuroprotective Effect of Stasis-Activating and Collaterals-Dredging Decoction on Focal Cerebral Ischemia-Reperfusion Injury in Rats

【目的】探讨化瘀通络汤对局灶性脑缺血-再灌注损伤大鼠的脑神经保护作用。【方法】将100只SD大鼠随机分为假手术组(蒸馏水灌胃),模型组(蒸馏水灌胃),化瘀通络汤低、中、高剂量组(7.65、15.30、30.60 g/kg对应灌胃),脑心通组(步长脑心通溶液0.864 g/kg灌胃)。各组大鼠连续灌胃7 d后,除假手术组,其余各组大鼠采用改良线栓法制备大脑中动脉栓塞模型。于造模后24 h采用改良神经功能缺损评分(mNSS)评估神经功能。处死大鼠后,采用2,3,5-氯化三苯基四氮唑(TTC)染色法测定脑梗死范围,苏木素-伊红(HE)染色法观察大鼠脑组织病理损伤变化,甲苯胺蓝染色法观察大鼠脑组织神经细胞形态及受损情况,原位末端转移酶标记(TUNEL)染色法观察大鼠神经元细胞凋亡情况,免疫组织化学法观察大鼠脑组织碱性成纤维细胞生长因子(bFGF)蛋白表达,酶联免疫吸附分析(ELISA)法检测大鼠血清炎性因子白细胞介素6(IL-6)表达水平。【结果】与假手术组比较,模型组大鼠神经功能缺损评分升高(P<0.05),脑梗死体积增大(P<0.05),大脑组织出现明显病理损伤,神经细胞凋亡率增加(P<0.05),梗死区域b FGF蛋白表达水平降低(P<0.05),血清中IL-6表达水平升高(P<0.05);与模型组比较,化瘀通络汤各剂量组上述指标得到明显改善,效果与脑心通组相当。【结论】化瘀通络汤对局灶性脑缺血-再灌注损伤大鼠有神经保护作用。

Objective To explore the neuroprotective effect of Stasis-Activating and Collaterals-Dredging Decoction(SCD)on the focal cerebral ischemia-reperfusion injury in rats. Methods One-hundred rats were randomly divided into sham-operation group(distilled water by gavage),model group(distilled water by gavage),low-,middle-and high-dose SCD groups(7.65,15.30,30.60 g·kg^(-1)·d^(-1) of SCD by gavage,respectively),Naoxintong group(0.864 g·kg^(-1)·d^(-1) of Buchang Naoxintong solution by gavage). After intragastric administration for 7 continuous days, the rat model of middle cerebral artery occlusion(MCAO)was established by modified thread-occlusion method. Twenty-four hours after modeling,the neurological function was assessed by modified neurological severity score(mNSS). After cervical luxation of rats,the infarct area of brain was measured by 2,3,5-triphenyltetrazolium chloride(TTC) staining, the pathological changes of brain tissue were observed by hematoxylin-eosin(HE)staining,the morphological characteristics of nerve cells were observed by toluidine blue staining,the cell apoptosis was observed by terminal deoxynucleotidyl transferase(TdT)-mediated dUTP-biotin nick end labeling(TUNEL)staining,the protein expression level of basic fibroblast growth factor(bFGF)in brain tissue was detected by immunohistochemistry,and the level of inflammatory factor interleukin 6(IL-6)in serum was detected by enzyme-linked immune-sorbent assay(ELISA). Results Compared with the sham-operation group,the mNSS was increased in the model group(P< 0.05),and the cerebral infarction volume was enhanced(P< 0.05),the obvious pathological injury in rat brain tissue was seen,the rate of neuronal apoptosis was raised(P< 0.05),the expression level of bFGF in the infarct area was lowered(P< 0.05),and the level of IL-6 in serum was increased(P< 0.05). Compared with the model group,the above indexes were improve in the various dose SCD groups, the effect being equivalent to that of the Naoxintong group. Conclusion SCD has neuroprotective effects on the focal cerebral ischemia-reperfusion injury in rats.