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目的研究脑络欣通对大鼠脑缺血再灌注损伤神经保护的作用机制。方法雄性SD大鼠144只,随机分为假手术组、模型组、对照组、治疗组,每组36只,各组随机数字表法分成1、3、7 d组,每个时间节点12只。复制脑缺血再灌注(MCAO/R)大鼠模型。治疗组造模后分别于1、3、7 d给予脑络欣通,对照组造模后分别1、3、7 d给予通心络。脑络欣通配制成浓度为0.26 g/m L的溶液,通心络配制成浓度为0.02 g/m L的溶液,均按10 m L/(kg·d)灌胃给药。假手术组及模型组分别1、3、7 d予以等量生理盐水灌胃。采用激光多普勒血流仪(LDF)测定大鼠缺血侧局部脑血流量(r CBF)。解剖大鼠取出脑组织并分离海马,采用HE染色法检测大鼠海马DG区;采用实时荧光定量PCR技术(RT-PCR)测定miRNA9a、miRNA15a、miRNA124的表达;采用Western blot法检测Wnt3a、Wnt5a、β-catenin的表达。结果与假手术组比较,模型组大鼠脑组织明显缺血,脑血流量显著降低(P<0.01),海马中miRNA9a、miRNA15a表达均显著降低(P<0.01),miRNA124显著增高(P<0.01),Wnt3a、Wnt5a、β-catenin表达显著增高(P<0.01);与模型组比较,脑络欣通1、3、7 d组大鼠脑血流量恢复明显,海马中miRNA9a、miRNA15a表达显著增高(P<0.01),miRNA124表达水平显著降低(P<0.01),Wnt3a、Wnt5a、β-catenin表达显著增高(P<0.01)。结论脑络欣通对于MCAO/R大鼠脑损伤有明显疗效,可通过提高局部脑血流量,提高海马miRNA9a、miRNA15a表达,降低海马miRNA124表达,提高Wnt3a、Wnt5a、β-catenin的表达,起到抑制缺血再灌注对大鼠脑组织损伤的作用。
Objective To study the mechanism of Naoluoxintong’s neuroprotection against cerebral ischemia-reperfusion injury in rats. Methods A total of 144 male Sprague-Dawley rats were randomly divided into sham-operation group, model group, control group and treatment group, with 36 rats in each group. The rats in each group were divided into 1, . Rat models of cerebral ischemia-reperfusion (MCAO / R) replication. After treatment, the rats in the treatment group were given Naoxintong on the 1st, 3rd, 7th day respectively, while the control group were given Tongxinluo on the 1st, 3rd, 7th day respectively. Naoluo Tong Tong made a concentration of 0.26 g / m L solution, Tongxinluo formulated to a concentration of 0.02 g / m L of the solution, according to 10 m L / (kg · d) intragastric administration. Sham-operation group and model group were given the same amount of normal saline 1,3,7 d gavage. The ischemic regional cerebral blood flow (r CBF) was measured by laser Doppler flowmetry (LDF). The hippocampus were isolated from the hippocampus of rats by HE staining and the expression of miRNA9a, miRNA15a and miRNA124 was detected by real-time fluorescence quantitative PCR (RT-PCR). The expressions of Wnt3a, Wnt5a, β-catenin expression. Results Compared with the sham operation group, the brain tissue of rats in the model group was significantly ischemia and the cerebral blood flow was significantly decreased (P <0.01). The expression of miRNA9a and miRNA15a in the hippocampus was significantly decreased (P <0.01), while the miRNA124 was significantly increased ), Wnt3a, Wnt5a, and β-catenin were significantly increased (P <0.01). Compared with the model group, the cerebral blood flow of rats in the 1, 3 and 7 d groups of Naoxintong group recovered obviously. The expressions of miRNA9a and miRNA15a in hippocampus were significantly increased (P <0.01). The expression of miRNA124 was significantly decreased (P <0.01) and the expression of Wnt3a, Wnt5a and β-catenin was significantly increased (P <0.01). CONCLUSION: Naoluo Xintong has obvious curative effect on brain injury in MCAO / R rats. It can improve the expression of miRNA9a and miRNA15a in hippocampus, decrease the expression of miRNA124 and increase the expression of Wnt3a, Wnt5a and β-catenin in hippocampus by increasing local cerebral blood flow, Inhibitory effects of ischemia / reperfusion on brain injury in rats.