益气熄风胶囊对大鼠局灶性脑缺血再灌注损伤的保护作用

Protective effect of Yiqi Xifeng Capsules on focal cerebral ischemia reperfusion injury in rats

目的研究益气熄风胶囊防治大鼠局灶性脑缺血再灌注损伤的作用以及其作用机制。方法将SD大鼠随机分为假手术组、模型组、尼莫地平片(30 mg/kg)组和益气熄风胶囊0.771 8、1.543 5、3.087 0、6.174 0 g/kg组,每个组再分为脑梗死率检测、病理学研究2个亚组。益气熄风胶囊于造模前ig给药,1次/d,连续7 d;假手术组和模型组给予生理盐水,给药体积均为10 m L/kg。于第7天给药后30 min制造大脑中动脉闭塞(MCAO)模型。大脑中动脉阻断3 h后,进行再灌注。假手术组仅在动脉处备线处理,待给予生理盐水后去线即可。缺血再灌注后24 h后TTC染色法测定脑梗死体积,计算脑梗死率。缺血再灌注24 h时计算脑指数和脑含水量。各组大鼠在脑缺血再灌注后3、24 h进行神经功能评分。采用苏木素、伊红进行染色,光镜下观察各组大鼠脑组织的病理形态。采用甲苯胺蓝尼氏染色法检测尼氏小体表达情况,观察脑组织尼氏小体数量;采用免疫组化染色SP法测定i NOS蛋白的平均吸光度值,观察大鼠脑组织i NOS蛋白表达情况。结果益气熄风胶囊1.543 5、3.087 0 g/kg对局灶性脑缺血再灌注引起大鼠脑梗死率增大有明显降低作用(P<0.05),而益气熄风胶囊0.771 8、6.174 0 g/kg组大鼠脑梗死率无明显变化。益气熄风胶囊0.771 8、1.543 5、3.087 0、6.174 0 g/kg组大鼠脑指数和脑含水量无明显变化。造模后3 h益气熄风胶囊0.771 8、3.087 0、6.174 0 g/kg神经行为学评分明显增加(P<0.05、0.01),对局灶性脑缺血再灌注而引起大鼠神经功能损伤症状有改善作用,而益气熄风胶囊1.543 5 g/kg的作用不明显。与模型组比较,益气熄风胶囊1.543 5 g/kg组大鼠脑组织的灰质病变、白质病变、脑膜、室管膜病变均明显减轻(P<0.01),尼氏染色计数平均吸光度值显著增加(P<0.01),脑组织中i NOS蛋白表达水平有降低的趋势,但是不具有统计学意义。结论益气熄风胶囊对大鼠局灶性脑缺血再灌注损伤具有明确的保护作用,其作用机制可能与脑组织病理病变减轻和尼氏小体数量增加有关。

Objective To investigate the protective effect of Yiqi Xifeng Capsules on cerebral ischemia reperfusion injury in rats and its mechanisms.Methods SD rats were randomly divided into Sham group, model group, Nimodipine Tablets （30 mg/kg） group, and Yiqi Xifeng Capsules 0.7718, 1.5435, 3.0870, and 6.1740 g/kg groups, and each group was further divided into two subgroups for cerebral infarction rate detection and pathology. Yiqi Xifeng Capsules was ig administered before the model preparation, 1 time/d, continuous treatment for 7 d. Sham group and the model group were given normal saline, and the volume of administration was 10 mL/kg. Middle cerebral artery occlusion （MCAO） models were prepared at 30 min after 7 d of administration. The middle cerebral artery was blocked for 3 h after reperfusion. In Sham group, the line was prepared only in the artery, and after normal saline was given, the line would be removed. After ischemia reperfusion for 24 h, the volume of cerebral infarction was measured by TTC staining, and the cerebral infarction rate was calculated. The brain index and brain water content were calculated at 24 h after ischemia reperfusion. Neurological function scores were measured at 3 and 24 h after cerebral ischemia reperfusion in rats. Hematoxylin and eosin were used to stain, and the pathological morphology of rat brain tissue was observed under light microscope. Nissl body expression was determined by toluidine blue Nissl staining method, and numbers of Nissl bodies were observed. The average absorbance of iNOS protein was determined by immunohistochemical staining and SP method, and the expression of iNOS protein in brain tissue of rats was observed.Results Yiqi Xifeng Capsules （1.5435 and 3.0870 g/kg） could significantly reduce the cerebral infarction rate of focal cerebral ischemia reperfusion injury in rat （P 〈 0.05）, while 0.7718 and 6.1740 g/kg had no significant effect. There was no significant change in brain index and brain water content in rats treated by Yiqi Xifeng Capsule （0.7718, 1.5435, 3.0870, and 6.1740 g/kg）. After model for 3 h, neurobehavioral scores of Yiqi Xifeng Capsules （0.7718, 3.0870 and 6.1740 g/kg） group were increased significantly （P 〈 0.05, 0.01）, and focal cerebral ischemia reperfusion in rats caused by nerve function damage symptoms were improved, while Yiqi Xifeng Capsules 1.5435 g/kg group had no obvious effect. Compared with model group, the pathological changes of gray matter, white matter, meninges and ependymal lesions in the brain tissue of rats in Yiqi Xifeng Capsules （1.5435 g/kg） group were significantly alleviated （P 〈 0.01）, the average absorbance value of Nissl staining were significantly increased （P 〈 0.01）, and expression level of iNOS protein in brain tissue tended to decrease, but it was not statistically significant. Conclusion Yiqi Xifeng Capsules have definite protective effect on focal cerebral ischemia reperfusion injury in rats, which may be related to the decrease of pathological changes of brain tissue and the increase of Nissl bodies number.