银星养脑片对血管性痴呆模型大鼠的记忆改善作用及机制研究

Study on Effect of Yinxing Yangnao Tablet on Memory Improvement of Vascular Dementia Model Rat and Its Mechanism

目的初步探讨银星养脑片治疗血管性痴呆的药效及可能机制。方法构建双侧颈动脉结扎的血管性痴呆大鼠模型,术后14 d分别给予银星养脑片0.07,0.135,0.27 g·kg^(−1)及吡拉西坦片0.4 g·kg^(−1),持续灌胃14 d后进行水迷宫行为学测试,随后收集血清及动物脑组织,检测血清中炎症因子TNF-α、IL-1β含量,对脑组织进行HE染色及尼氏染色观察海马区病理结构变化。使用BV2细胞构建LPS刺激模型,给予银星养脑片0.5,1,2 mg·kg^(−1)不同剂量后检测细胞上清炎症相关因子NO、IL-6、TNF-α的释放量,收集细胞蛋白检测iNOS、NF-κB p65、p-NF-κB p65、IκB-α、p-IκB-α、NLRP3、caspase-1炎症相关蛋白表达变化,免疫荧光检测细胞iNOS、NLRP3的表达情况。结果水迷宫结果提示血管性痴呆大鼠出现明显的记忆损伤,而银星养脑片给药组能有效改善记忆损伤情况,且还能降低血管性痴呆大鼠血清炎症水平;在BV2细胞上给予银星养脑片的干预能减少LPS刺激后的炎症因子释放,抑制NF-κB p65和IκB-α的磷酸化蛋白的增加,抑制NF-κB下游NLRP3/caspase-1通路的激活,有效地起到抗炎的作用。结论银星养脑片能有效改善血管性痴呆大鼠的记忆损伤,有可能通过抑制炎症相关NF-κB/NLRP3/caspase-1通路起到抗炎的作用。

OBJECTIVE To preliminary explore the efficacy and possible mechanism of Yinxing Yangnao Tablets in the treatment of vascular dementia.METHODS A rat model of vascular dementia with bilateral carotid artery ligation was constructed.Yinxing Yangnao Tablets 0.07,0.135,0.27 g·kg^(−1)and piracetam tablets 0.4 g·kg^(−1)were given 14 d after the operation.After 14 d of continuous gavage,morris water maze tested,then serum and brain tissue collected to detect serum inflammatory factors TNF-αand IL-1βconcentration,and the pathological structure of hippocampus observed by HE and Nissl staining.BV2 cells were used to construct an LPS stimulation model,and Yinxing Yangnao Tablets were given different doses of 0.5,1,2 mg·kg^(−1)to detect the release of inflammation-related factors NO,IL-6 and TNF-αin the cell supernatant,and collect cell protein detection iNOS,NF-κB p65,p-NF-κB p65,IκB-α,p-IκB-α,NLRP3,caspase-1 inflammation-related protein expression changes,immunofluorescence detection of cell iNOS,NLRP3 expression.RESULTS The results of the morris water maze indicated that the rats with vascular dementia had obvious memory impairment,and the Yinxing Yangnao Tablets administration group could effectively improve the memory impairment and also reduce the serum inflammation level in the rats with vascular dementia.At the same time,Yinxing Yangnao Tablets intervention could reduce the release of inflammatory factors after LPS stimulation on BV2 cells,inhibit the increase of NF-κB p65 and IκBαphosphorylation protein,and inhibit the activation of the NLRP3/caspase-1 pathway downstream of NF-κB,effectively acting as an anti-inflammatory agent.CONCLUSION Yinxing Yangnao Tablets can effectively improve the memory impairment of rats with vascular dementia,and may have anti-inflammatory effects by inhibiting inflammation-related NF-κB/NLRP3/caspase-1 pathway.