脉络宁对大鼠脑缺血再灌注损伤的保护作用及其机制研究

Protective effect of Mailuoning injection on cerebral ischemia/reperfusion injury in rats and its mechanism

目的:探讨脉络宁注射液对大鼠脑缺血再灌注(ischemia/reperfusion,I/R)损伤的保护作用及其机制。方法:健康成年雄性Sprague-Dawley(SD)大鼠随机分为假手术组、模型组、依达拉奉对照组(3 mg·kg^(-1))、脉络宁高、中、低剂量组(4,2,1 mL·kg^(-1)),每组10只,尾静脉注射给药。采用线栓法阻断大脑中动脉(middle cerebral artery occlusion,MCAO)建立大鼠I/R模型。缺血2 h,再灌注24 h时后,HE染色观察大鼠缺血侧脑组织神经血管单元(neurovascular unit,NVU)的病理变化,免疫组织化学法检测胶质纤维酸性蛋白(glialfibrillary acidic protein,GFAP)及离子钙接头蛋白(ionized calcium binding adaptor molecule 1,Iba1)的表达,蛋白免疫印记法检测肿瘤坏死因子-α(tumornecrosis factor-α,TNF-α)、白细胞介素-1β(interleukin1β,IL-1β)、血管细胞黏附分子-l(vascular cell adhesion molecule-1,VCAM-1)、细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)的表达。结果:脉络宁注射液明显改善缺血再灌注大鼠脑组织NVU的病理变化;减少GFAP和Iba1阳性细胞的数量;显著降低缺血再灌注大鼠脑组织TNF-α,IL-1β,ICAM-1,VCAM-1的表达。结论:脉络宁注射液对脑缺血再灌注大鼠神经血管单元具有明显的保护作用,其机制可能涉及抑制星形胶质细胞和小胶质细胞的激活,以及多种炎性因子的分泌和表达。

Objective： To discuss the protective effect of Mailuoning injection on ischemia/reperfusion （I/R） injury in rats and its mechanism. Method： Healthy male adult Sprague-Dawley （SD） rats were randomly divided into the sham operation group, the model group, the edaravone （3 mg . kg- 1 ） control group, and Mailuoning high, middle and low-dose groups （4, 2, 1 mL . kg- 1 ）, with 10 rats in each group, and administered with drugs through tail intravenous injection. The middle cerebral artery occlusion （MCAO） was adopted to establish the rat ischemia/reperfusion model. After the ischemia for 2 h and reperfusion for 24 h, the pathological changes in neurovascular units （NVU） of brain tissues at the ischemia side was observed by HE staining. The expressions of glialfibrillary acidic protein （GFAP） and ionized calcium-binding adaptor molecule 1 （Ibal） were detected by the immunohistochemical method. The expressions of tumor necrosis factor-a （TNF-a）, interleukin 1β （IL-1β）, vascular cell adhesion molecule-1 （VCAM-1） and intercellular adhesion molecule-1 （ICAM-1） were detected by the western blotting technique. Result： Mailuoning injection could significantly improve the pathological changes in cortical penumbra brain tissue UVN of （I/R） rats, reduce the number of GFAP and Ibal positive cells, and significantly decrease the expressions of TNF-a, IL-1β, VCAM-1 and ICAM-1 of brain tissues of I/R rats. Conclusion： Mailuoning injection shows an obvious protective effect on UVN of I/R rats. Its mechanism may involve the inhibition of the activation of astrocyte and microglia and the secretion and expression of various inflammatory factors.