癫痫清颗粒对阿尔茨海默病小鼠模型细胞焦亡影响

Effects of Dianxianqing Granules(癫痫清颗粒)on Pyroptosis in Alzheimer’s Disease Model Mice

目的 研究阿尔茨海默病(Alzheimer’s disease,AD)模型小鼠神经细胞焦亡受癫痫清颗粒的作用效果。方法先将小鼠随机分为假手术组、模型组、盐酸多奈哌齐组(1.3 mg/kg)、NLRP3炎症小体拮抗剂(MCC950)组、癫痫清颗粒组(12.48 g/kg)和MCC950+癫痫清颗粒组。除假手术组外,其余各组小鼠的左侧侧脑室内均注射淀粉样蛋白(Aβ25-35)制作AD小鼠模型,癫痫清颗粒连续给药21 d,1次/d。第16~21天,Morris水迷宫检测小鼠的空间学习和记忆能力,第22天各组小鼠取材,通过HE染色对神经元病理学变化进行观察,ELISA检测脑组织中白介素-1β(interleukin-1β,IL-1β)、白介素-18(interleukin-18,IL-18)及肿瘤坏死因子-α(TNF-α)蛋白表达水平,免疫荧光检测脑组织中NLRP3蛋白表达水平,Western blot检测海马组织中NLRP3、IL-1β、IL-18、TNF-α、凋亡相关斑点样蛋白(ASC)及半胱天冬氨酸酶(Caspase-1)蛋白表达水平。结果 与假手术组比较,模型组小鼠造模后游泳潜伏期显著增加,穿越平台次数大幅度减少,神经元胞体形状不规则,排列紊乱,间隙增宽,出现变性和坏死,NLRP3、IL-1β、IL-18、ASC、TNF-α及Caspase-1蛋白表达增加。与模型组相比较,各给药组通过训练后,游泳潜伏期明显缩短,穿越平台次数逐步增加;神经元细胞组织结构完整,排列紧密,间隙减小;NLRP3、IL-1β、IL-18、ASC、TNF-α及Caspase-1蛋白在组织中的表达水平均显著降低。结论 癫痫清颗粒通过抑制NLRP3的活化从而抑制神经焦亡的发生,同时使AD模型小鼠的学习和记忆能力得到改善。

Objective To study the effect of Dianxianqing Granules(癫痫清颗粒)on neuronal pyroptosis in Alzheimer’s disease(AD)model mice.Methods Sham operation group underwent sham operation,and AD model was induced by intracerebroventricular(ICV)injection ofβ-amyloid 25-35(Aβ25-35),mice were randomly divided into sham operation group,model group,donepezil hydrochloride group(1.3 mg/kg),NLRP3 inflammasome antagonist(MCC950)injection group,Dianxianqing Granules group(12.48 g/kg)and MCC950+Dianxianqing Granules group.Administration groups were given relevant drug solution intrasgastrcially once a day,for consecutive 21 days.At last administration,the learning and memory ability was determined by morris water maze test,and HE staining was used to observe the pathological changes of neurons,and IL-1β,IL-18 and TNF-αexpression in brain tissue were measured with enzyme linked immunosorbent assay.Immunofluorescence was used to detect the protein expression of NLRP3 in brain tissue.Western blotting assay was used to determine the protein expressions of NLRP3,IL-1β,IL-18,TNF-α,ASC and Caspase-1 in brain tissue.Results Compared with sham operation group,latency and protein expressions of NLRP3,IL-1β,IL-18,ASC,TNF-αand Caspase-1 in brain were increased significantly in model group;platform crossing times was decreased significantly,there were obvious pathological morphological changes such as irregular shape of neuron cell body,disordered arrangement,widening of gap,degeneration and necrosis in brain tissue.Compared with model group,latency and protein expressions of NLRP3,IL-1β,IL-18,ASC,TNF-αand Caspase-1 in brain were decreased significantly in Dianxianqing Granules,while the platform crossing times was increased significantly,pathological changes of brain tissue were improved to different extents in administration groups.Conclusion Dianxianqing Granules can further inhibit the occurrence of neuronal pyroptosis by inhibiting the activation of NLRP3,and improve the learning and memory ability of AD model mice.