加味不忘散对AD模型大鼠海马区NLRP3炎症通路中相关因子表达的影响

Effect of Modified Buwangsan on Expression of Related Molecules in NLRP3 Inflammatory Signaling Pathway in Hippocampus of AD Rats

目的:通过观察加味不忘散对阿尔茨海默病(AD)模型大鼠学习记忆能力及海马区NOD样受体热蛋白结构域3(NLRP3)炎症通路中相关分子NLRP3,天冬氨酸蛋白酶-1(Caspase-1)和白细胞介素-1β(IL^-1β)表达的影响,探讨其作用机制。方法:通过Morris水迷宫筛选出合格SD大鼠52只,随机均分为假手术组,AD模型组,加味不忘散低剂量组(1.5 g·kg^-1),加味不忘散高剂量组(3 g·kg^-1)。采用双侧海马注射β-淀粉样蛋白1-42(Aβ1-42)5μL(2 g·L^-1)建立AD模型大鼠。造模后分别予低、高剂量加味不忘散灌胃,每日1次,连续4周。灌胃结束后通过Morris水迷宫法检测大鼠学习记忆能力,判断造模是否成功;通过实时荧光定量聚合酶链式反应(Real-time PCR),蛋白免疫印迹法(Western blot)检测大鼠海马组织中NLRP3,Caspase-1,IL^-1βmRNA和蛋白的表达水平。结果:与假手术组比较,AD模型组大鼠学习记忆能力明显下降(P<0.05);与AD模型组比较,加味不忘散低剂量组大鼠学习记忆能力无改善,NLRP3,Caspase-1,IL^-1βmRNA和蛋白的表达水平均无统计学差异,加味不忘散高剂量组大鼠学习记忆能力明显改善,NLRP3,Caspase-1,IL^-1βmRNA和蛋白的表达均明显下降(P<0.05)。结论:高剂量加味不忘散可改善大鼠学习记忆能力,其机制可能与下调NLRP3炎症通路中NLRP3及下游Caspase-1,IL^-1β的表达,抑制海马组织内的炎症反应有关。

Objective:To investigate the effects of modified Buwangsan on the learning and memory ability of Alzheimer’s disease(AD)model rats and the expression of NOD-like receptor 3(NLRP3),cysteinecontaining aspartate-specific proteases 1(Caspase-1)and interleukin-1 beta(IL^-1β)in NLRP3 inflammatory pathway in hippocampus of AD model rats,and exploring the underlying mechanism of modified Buwangsan.Method:The 52 eligible rats were randomly divided into sham control group,AD model group,low-dose modified Buwangsan group(1.5 g·kg^-1)and high-dose modified Buwangsan group(3 g·kg^-1).AD mouse model was established by bilateral hippocampus injection of Aβ1-425μL(2 g·L^-1).The rats in low-dose and high-dose modified Buwangsan group received low and high dose modified Buwangsan respectively within the next 4 weeks,once daily.The learning and memory ability was tested by Morris water maze.The expression of NLRP3,Caspase-1 and IL^-1βmRNA was tested by quantitative PCR(Real-time PCR)and Western blot.Result:As compared with the sham group,the learning and memory ability of the rats were significantly impaired(P<0.05).Compared with AD model group,the learning and memory ability and the expression levels of NLRP3,Caspase-1,and IL^-1βmRNA and protein were all no statistical differences in low-dose modified Buwangsan group,while the learning and memory ability of the rats were significantly improved and the expression of NLRP3,Caspase-1 and IL^-1βmRNA in hippocampus of rats was significantly decreased in high-dose modified Buwangsan group(P<0.05).Conclusion:High-dose modified Buwangsan could attenuate neuroinflammation in the hippocampus of AD mouse model via inhibiting the expression of NLRP3,Caspase-1 and IL^-1β,which may be the mechanisms of modified Buwangsan could be used to ameliorate the learning and memory ability of AD mouse model.