锁阳黄酮对阿尔茨海默病大鼠海马组织ROS含量及NADPH氧化酶、NLRP3表达的影响  

Effects of Cynomorium songaricum Flavonoids on ROS content and Expressions of NADPH Oxidase and NLRP3 in Hippocampus of Alzheimer Disease Rats

作　　者：吕鑫 顾志荣[2] 祁梅[2] 郭燕 毛小文 葛斌[2] LYU Xin;GU Zhirong;QI Mei;GUO Yan;MAO Xiaowen;GE Bin(College of Pharmacy,Gansu University of Chinese Medicine,Lanzhou 730000,China;Gansu Provincial Hospital,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学药学院,甘肃兰州730000 [2]甘肃省人民医院,甘肃兰州730000

出　　处：《中国中医药信息杂志》2023年第4期82-87,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：甘肃省自然科学基金(20JR5RA154);甘肃省青年科技基金(21JR7RA634);甘肃省人民医院青年项目(20GSSY4-29);“白求恩·求索-药学科研能力建设”项目(B-19-H-20200622);甘肃省中药质量与标准研究重点实验室开放基金(ZYZL18-004);甘肃省中药药理与毒理学重点实验室开放基金(ZDSYS-KJ-2018-010)。

摘　　要：目的 观察锁阳黄酮对β淀粉样蛋白(Aβ)_(1-42)诱导的阿尔茨海默病(AD)大鼠海马组织活性氧(ROS)含量及NADPH氧化酶、Nod样受体蛋白3(NLRP3)表达的影响,探讨其神经保护作用的机制。方法 70只SPF级Wistar雄性大鼠随机分为空白组、假手术组、模型组、多奈哌齐组及锁阳黄酮低、中、高剂量组,每组10只。大鼠于双侧海马注射Aβ_(1-42)建立AD大鼠模型。假手术组注射等体积生理盐水,空白组不予任何处理。多奈哌齐组和锁阳黄酮低、中、高剂量组予相应药物灌胃,空白组、假手术组和模型组予等体积生理盐水灌胃,连续28 d。Morris水迷宫实验评价大鼠学习记忆能力,HE染色观察海马组织病理变化,免疫组化检测海马组织Aβ_(1-42)表达,ELISA检测海马组织及血清ROS含量,Western blot检测海马组织NADPH氧化酶亚基(gp91^(phox)、p22^(phox)、p67^(phox)、p47^(phox))、NLRP3、白细胞介素(IL)-1β蛋白表达,RT-PCR检测海马组织NADPH氧化酶各亚基(gp91^(phox)、p22^(phox)、p67^(phox)、p47^(phox))mRNA表达。结果 与假手术组比较,模型组大鼠Morris水迷宫实验逃避潜伏期明显延长(P<0.01),跨平台次数和目标象限停留时间明显减少(P<0.01);海马CA1区锥体细胞凋亡明显,局部锥体细胞排列紊乱,海马组织Aβ_(1-42)表达明显升高(P<0.05);海马组织及血清ROS含量明显增加(P<0.01),海马组织NADPH氧化酶各亚基和NLRP3、IL-1β蛋白表达明显升高(P<0.01),NADPH氧化酶各亚基mRNA表达明显升高(P<0.05,P<0.01)。与模型组比较,锁阳黄酮高剂量组大鼠Morris水迷宫实验逃避潜伏期明显缩短(P<0.01),跨平台次数和目标象限停留时间明显增加(P<0.05,P<0.01);海马CA1区病理形态有所改善,海马组织Aβ_(1-42)表达明显降低(P<0.05);海马组织及血清ROS含量明显减少(P<0.01),海马组织NADPH氧化酶各亚基和NLRP3、IL-1β蛋白表达明显降低(P<0.05,P<0.01),NADPH氧化酶各亚基mRNA表达明显降低(P<0.05,Objective To investigate the effects of Cynomorium songaricum flavonoids(CSF) on reactive oxygen species(ROS) content,the expressions of NADPH oxidase and Nod-like receptor protein 3(NLRP3) in hippocampus of Aβ_(1-42)-induced Alzheimer disease(AD) rats.Methods Totally 70 SPF Wistar male rats were randomly divided into blank group,sham-operation group,model group,donepezil group,and CSF low-,medium-,and high-dosage groups,with 10 rats in each group.Aβ_(1-42)were injected into the bilateral hippocampus to establish AD rat model,the sham-operation group was injected with an equal volume of normal saline,the blank group did not receive any treatment.The donepezil group and CSF low-,medium-,and high-dosage groups were given corresponding drugs by gavage,the blank group,sham-operation group and model group were given equal volume of normal saline for 28 consecutive days.Morris water maze were used to test the learning and memory ability,HE staining was used to observe the pathological changes in hippocampus,the expression of Aβ_(1-42) in hippocampus was observed by immunohistochemistry,the contents of ROS in hippocampus and serum were detected by ELISA,the protein expressions of NADPH oxidase subunits(gp91^(phox),p22^(phox),p67^(phox),and p47^(phox)),NLRP3,and IL-1β in hippocampus were detected by Western blot,the expressions of NADPH oxidase subunit(gp91^(phox),p22^(phox),p47^(phox)and p67^(phox)) mRNA in hippocampus were detected by RT-PCR.Results Compared with the sham-operation group,the escape latency of Morris water maze test of rats in the model group significantly prolonged(P<0.01),the number of cross-platforms and the dwell time in the target quadrant significantly decreased(P<0.01);the apoptosis of pyramidal cells in hippocampal CA1 area was obvious,the arrangement of local pyramidal cells was disordered,and the expression of Aβ_(1-42)in hippocampus significantly increased(P<0.05);the content of ROS in hippocampus and serum significantly increased(P<0.01),the protein expressions of each subunit of NADPH

关 键 词：锁阳黄酮 阿尔茨海默病 Β淀粉样蛋白 NADPH氧化酶 活性氧 NLRP3 大鼠 

分 类 号：R285.5[医药卫生—中药学]