癫痫清颗粒改善AD模型小鼠血脑屏障损伤的作用研究  

Improvement effects of Dianxianqing granule on blood-brain barrier injury in Alzheimer’s disease model mice

作　　者：齐越 李昭[2] 侯侠 黄培池 贾冬 杨彩瑜 董笑博 房小楠 明彩荣 QI Yue;LI Zhao;HOU Xia;HUANG Peichi;JIA Dong;YANG Caiyu;DONG Xiaobo;FANG Xiaonan;MING Cairong(Dept.of Pharmaceutical Technology,Jiangsu Provincial Xuzhou Pharmaceutical Vocational College,Jiangsu Xuzhou 221116,China;Dept.of Pharmacology,College of Pharmacy,Shenyang Medical College,Shenyang 110031,China;Dept.of Pharmacology,the Second Affiliated Hospital of Liaoning University of TCM,Shenyang 110034,China)

机构地区：[1]江苏省徐州医药高等职业学校药学技术系,江苏徐州221116 [2]沈阳医学院药学院药理室,沈阳110031 [3]辽宁中医药大学附属第二医院药理室,沈阳110034

出　　处：《中国药房》2022年第9期1062-1067,共6页China Pharmacy

基　　金：“重大新药创制”科技重大专项子课题(No.2012ZX09102-201-005);徐州市科技局课题(No.KC20146)。

摘　　要：目的研究癫痫清颗粒通过调控核苷酸结合域样受体蛋白3(NLRP3)炎症小体信号通路对阿尔茨海默病(AD)模型小鼠血脑屏障(BBB)损伤的改善作用。方法将125只小鼠按体质量随机分为假手术组(n=25)和造模组(n=100)。造模组小鼠采用侧脑室注射β-淀粉样蛋白25~35的方法复制AD模型;假手术组小鼠同法注射生理盐水。选取造模成功的100只小鼠,按体质量随机分为模型组、盐酸多奈哌齐片组(阳性对照1,1.3 mg/kg,灌胃给药)、MCC950组[阳性对照2(选择性NLRP3抑制剂),10 mg/kg,腹腔注射给药]和癫痫清颗粒组(以生药总量计12.48 g/kg,灌胃给药),每组25只。造模后第2天,各给药组小鼠给予相应药物,每天1次,连续21 d;假手术组和模型组小鼠灌胃水并腹腔注射生理盐水。末次给药后,采用Y迷宫实验检测小鼠的学习记忆能力,采用伊文思蓝渗漏实验测定小鼠BBB通透性;检测小鼠脑组织中NLRP3、离子钙接头蛋白(IBA-1)、核转录因子κB(NF-κB)p65、p53上调凋亡调控因子(PUMA)、咬合蛋白(ocln)、闭锁小带蛋白1(ZO-1)、紧密连接蛋白5(cldn5)的表达水平。结果与模型组比较,各给药组小鼠的自发交替反应率和脑组织中ocln、cldn5、ZO-1蛋白表达水平均显著升高(P<0.05或P<0.01),脑组织内的伊文思蓝含量及NLRP3、IBA-1、PUMA、NF-κB p65蛋白表达水平均显著降低(P<0.05或P<0.01)。结论癫痫清颗粒可通过抑制NLRP3炎症小体信号通路起到改善AD模型小鼠BBB损伤的作用。OBJECTIVE To study the improvement effects of Dianxianqing granule on blood-brain barrier(BBB)injury in Alzheimer’s disease(AD)model mice by regulating NLR family pyrin domain containing 3(NLRP3)inflammasome signaling pathway.METHODS Totally 125 mice were randomly divided into sham operation group(n=25)and modeling group(n=100)by body weight.AD model was induced by intracerebroventricular injection ofβ-amyloid 25-35 in model group.Sham operation group was given normal saline with same method.The 100 model mice were randomly divided into model group,Donepezil hydrochloride tablets group(positive control 1,1.3 mg/kg,i.g.),MCC950 group[positive control 2(selective NLRP3 inhibitor),10 mg/kg,i.p.]and Dianxianqing granule group(12.48 g/kg by crude drug,i.g.)by body weight,with 25 mice in each group.Second day after modeling,administration groups were given relevant medicine,once a day,for consecutive 21 d.Sham operation group and model group were given intragastric administration of water and intraperitoneal injection of normal saline.At last administration,the learning and memory ability was determined by Y maze test,and blood-brain barrier permeability was measured by Evans blue leakage assay.The expressions of NLRP3,anti-ionized calcium-binding adapter molecule 1(IBA-1),nuclear factor kappa B(NF-κB)p65,p53 upregulated modulator of apoptosis(PUMA),occludin(ocln),zonula occluden-1(ZO-1)and claudin-5(cldn5)in cerebral tissue were determined.RESULTS Compared with model group,spontaneous alternate response rate,protein expressions of ocln,cldn5 and ZO-1 in cerebral tissue were increased significantly in administration groups(P<0.05 or P<0.01);Evans blue content and protein expressions of NLRP3,IBA-1,PUMA and NF-κB p65 in cerebral tissue were decreased significantly(P<0.05 or P<0.01).CONCLUSIONS Dianxianqing granule can improve BBB injury of AD model mice by inhibiting NLRP3 inflammasome signaling pathway.

关 键 词：癫痫清颗粒 阿尔茨海默病 血脑屏障 核苷酸结合域样受体蛋白3炎症小体 紧密连接蛋白 

分 类 号：R965[医药卫生—药理学]