酸枣仁汤防治睡眠剥夺性大鼠学习记忆功能变化及其对NLRP3通路的影响

Effect of Suanzaoren Tang on Learning-memory Function and NLRP3 Pathway in Sleep-deprived Rats

目的:基于Nod样受体蛋白3(NLRP3)炎性小体通路探讨酸枣仁汤改善睡眠剥夺大鼠学习记忆功能的作用机制。方法:将实验大鼠随机分为正常组,模型组,艾司唑仑组(5.4×10^(-4)g·kg^(-1)·d^(-1)),酸枣仁汤低剂量组(4.59 g·kg^(-1)·d^(-1)),酸枣仁汤高剂量组(18.36 g·kg^(-1)·d^(-1))。除正常组外,其他组构建睡眠剥夺动物模型,造模与给药同时进行,连续给药30 d。应用水迷宫评价小鼠学习记忆力;应用实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测小鼠海马中Nod样受体蛋白3(NLRP3),半胱天冬酶募集结构域的凋亡相关斑点样蛋白(ASC),天冬氨酸特异性半胱氨酸蛋白酶-1(Caspase-1),白细胞介素-1β(IL-1β),白细胞介素-18(IL-18)m RNA和蛋白表达水平。结果:与正常组比较,模型组大鼠上平台潜伏期,游泳总路程和第1次抵原平台时间则显著增加(P<0.01),穿越平台次数和目标象限时间减少(P<0.01);与模型组比较,酸枣仁汤低剂量组和酸枣仁汤高剂量组大鼠上平台潜伏期,游泳总路程和第1次抵原平台时间不同程度减少(P<0.05,P<0.01),穿越平台次数和目标象限时间明显增加(P<0.05,P<0.01),艾司唑仑组变化不显著。与正常组比较,模型组大鼠海马中NLRP3,ASC,Caspase-1,IL-1β,IL-18 m RNA和蛋白表达量明显升高(P<0.05,P<0.01);与模型组大鼠比较,酸枣仁汤低剂量组和酸枣仁汤高剂量组大鼠海马中NLRP3,ASC,Caspase-1,IL-1β,IL-18 mRNA表达量均有不同程度降低(P<0.05),艾司唑仑组大鼠海马中NLRP3,ASC,Caspase-1,IL-1β,IL-18 mRNA表达量也有所减少,但差异无明显统计学意义。结论:酸枣仁汤可改善睡眠剥夺大鼠学习记忆功能,其机制与抑制海马NLRP3炎性小体通路,减轻神经炎性相关。

Objective:To explore the mechanism of Suanzaoren Tang in improving learning-memory of sleep-deprived rats based on Nod-like receptor 3(NLRP3)inflammatome pathway.Method:The rats were randomly divided into normal control group,model group,Eszolam group(5.4×10^(-4)g·kg^(-1)·d^(-1)),low-dose Suanzaoren Tang group(4.59 g·kg^(-1)·d^(-1))and high-dose Suanzaoren Tang group(18.36 g·kg^(-1)·d^(-1)).In addition to normal control group,other groups were used to constructed sleep-deprived model,which was concurrent with30-day continuous drug administration.Water maze was used to evaluate the learning-memory function of rats;The m RNA and protein expressions of NLRP3,apoptosis-related speckle proteins(ASC),aspartic acid-specific cysteine protease-1(Caspase-1),interleukin-1(IL-1)and IL-18 in the hippocampus of rats were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot.Result:Compared with control group,the incubation period of the platform,the total distance of swimming and the duration of first reaching the platform in model group were significantly increased(P<0.01),while the number of platform crossings and the target quadrant time were decreased(P<0.01).Compared with the model group,the incubation period,total swimming distance and the duration of first reaching the platform in low-dose Suanzaoren Tang group and high-dose Suanzaoren Tang group were decreased to different degrees(P<0.05,P<0.01),while the number of platform crossings and the target quadrant time were increased significantly(P<0.05,P<0.01),but with no significant change in estazolam group.Compared with normal control group,m RNA and protein expressions of NLRP3,ASC,Caspase-1,IL-1β,IL-18 in the hippocampus of the model group were significantly increased(P<0.05,P<0.01).Compared with model group,m RNA and protein expressions of NLRP3,ASC,Caspase-1,IL-1β and IL-18 in the hippocampus of the rats in low-dose Suanzaoren Tang group and high-dose Suanzaoren Tang group were all decreased to different degrees(P<0.05).The mRNA and protein expressions of NLRP3,ASC,Caspase-1,IL-1β and IL-18 in the hippocampus of Suanzaoren group also decreased,but with no significant change.Conclusion:Suanzaoren Tang can improve the learning-memory function of sleep-deprived rats,and its mechanism is related to the inhibition of NLRP3 inflammatome pathway in hippocampus and the alleviation of neuroinflammation.