基于NF-κB通路探讨钩藤对大鼠蛛网膜下腔出血后早期脑损伤的影响

Based on NF-κB Pathway Exploring the Effect of Rhynchophylla on Early Brain Injury after Subarachnoid Hemorrhage in Rats

目的:探究钩藤影响大鼠蛛网膜下腔出血后早期脑损伤的分子机制。方法:将80只大鼠分为假手术组(Sham组)、模型组(SAH组)、钩藤低剂量组(LD组)、钩藤高剂量组(HD组),每组20只。采用枕大池注血法制作蛛网膜下腔出血模型,药物干预7d后评估各组大鼠的神经功能评分,测定脑组织含水量、基底动脉直径,TUNEL染色观察各组大鼠神经元凋亡情况,Western blotting检测脑组织中Caspase-1、NLRP3、NF-κB-p65的表达水平。ELISA法检测大鼠脑组织中炎症因子IL-1β、IL-6和IL-18表达水平。结果:与Sham组相比较,SAH组大鼠神经功能Garcia评分明显下降(P<0.05),脑组织含水量明显升高(P<0.05),基底动脉直径明显减小(P<0.05),脑细胞凋亡明显增加(P<0.05),IL-1β、IL-6和IL-18表达水平明显升高(P<0.05),脑组织NLRP3、Caspase-1、NF-κB-p65表达水平明显升高(P<0.05);与SAH相比较,LD、HD组大鼠的Garcia评分明显升高(P<0.05),脑组织含水量明显降低(P<0.05),基底动脉直径明显增加(P<0.05),脑细胞凋亡减少(P<0.05),IL-1β、IL-6和IL-18的表达水平明显降低(P<0.05),Caspase-1、NF-κB-p65、NLRP3表达明显降低(P<0.05),与LD组大鼠相比较,HD组大鼠的Garcia评分升高(P<0.05),脑组织的含水量明显降低(P<0.05),基底动脉直径明显增加(P<0.05),脑细胞凋亡减少(P<0.05),IL-1β、IL-6和IL-18的表达水平明显降低(P<0.05),NLRP3、Caspase-1、NF-κB-p65表达降低(P<0.05)。结论:钩藤能够减轻蛛网膜下腔出血后发生早期脑损伤大鼠脑组织的炎症反应,减轻脑水肿和细胞凋亡,扩张基底动脉,从而改善神经功能,可能是通过抑制NF-κB信号通路,降低NLRP3炎性小体的表达来实现的。

Objective:To investigate the molecular mechanism of the effect of rhynchophylla on early brain injury after subarachnoid hemorrhage in rats.Methods:80 rats were divided into sham surgery group,model group,low-dose group of rhynchophylla,and high-dose group of rhynchophylla,with 20 rats in each group.The subarachnoid hemorrhage model was established by injecting blood into the occipital cistern.After 7 days of drug intervention,evaluate the neurological function score of each group of rats,measure the water content of brain tissue,basal artery diameter,ELISA method was used to detect the inflammatory factor expression level of IL-1β,IL-6 and IL-18 in rat brain tissue,using Western blotting to detect the expression level of NLRP3,Caspase-1,NF-κB-p65,TUNEL staining was used to observe the apoptosis of neurons in each group of rats.Results:Compared with the sham group,the Garcia score in the model group was significantly reduced(P<0.05),the water content of brain tissue was significantly increased(P<0.05),the diameter of basal artery was significantly reduced(P<0.05),and brain cell apoptosis was significantly increased(P<0.05),the expression levels of IL-1β,IL-6 and IL-18 were significantly increased(P<0.05),and NLRP3,Caspase-1,NF-κB-p65 significantly increased(P<0.05).Compared with the model group,the Garcia scores of rats in the low-dose and high-dose groups of rhynchophylla were significantly increased(P<0.05),the water content of brain tissue was significantly reduced(P<0.05),the diameter of basal artery was significantly increased(P<0.05),brain cell apoptosis was reduced(P<0.05),and the expression levels of IL-1β,IL-6 and IL-18 were significantly reduced(P<0.05),while the expression of Caspase-1,NLRP3 and NF-κB-p65 was significantly reduced(P<0.05).Compared with the low-dose group of rats,the high-dose group of rats showed an increase in Garcia score(P<0.05),the water content of brain tissue was reduced(P<0.05),the diameter of basal artery was significantly increased(P<0.05),brain cell apoptosis was reduced(P<0.05),and the expression levels of IL-1β,IL-6 and IL-18 were significantly reduced(P<0.05),while the expression of NLRP3,Caspase-1,NF-κB-p65 decreased(P<0.05).Conclusion:Rhynchophylla can alleviate the inflammatory response of brain tissue of rats with early brain injury after subarachnoid hemorrhage,alleviate brain edema and cell apoptosis,dilate the basal artery,and improve neurological function,possibly,by inhibiting the NF-κB signaling pathway reducing the expression of NLRP3 inflammasome is achieved.