基于cAMP/BDNF/NMDA通路探讨银杏内酯B对砷暴露大鼠神经功能损伤的保护作用  

Based on the cAMP/BDNF/NMDA pathway to investigate the protective effect of ginkgolide B on the neurological damage of arsenic-exposed rats

作　　者：刘冬 刘熠 LIU Dong;LIU Yi(Department of Urological Neurology,Chongqing Seventh People's Hospital,Chongqing 400054,China)

机构地区：[1]重庆市第七人民医院泌尿神经外科,重庆400054

出　　处：《职业与健康》2024年第24期3334-3341,共8页Occupation and Health

基　　金：重庆市科卫联合医学科研项目(2019QNXM009)。

摘　　要：目的基于环磷酸腺苷(cyclic adenosine monophosphate,cAMP)/脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)/N-甲基-D-天冬氨酸(N-methyl-D-aspartic acid,NMDA)通路探讨银杏内酯B对砷暴露大鼠神经功能损伤的保护作用。方法40只SD大鼠分为正常对照组、砷暴露染毒组(15 mg/kg)、银杏内酯B低、高剂量组(200、400 mg/kg)。砷暴露染毒组、银杏内酯B低、高剂量组大鼠每日予以亚砷酸钠灌胃,持续8周;试验期间,银杏内酯B低、高剂量大鼠每日予以相应剂量的银杏内酯B灌胃,正常对照组、砷暴露染毒组给予等体积的生理盐水灌胃。试验结束后,莫里斯水迷宫评价大鼠空间学习和记忆能力、神经严重程度评分表评价大鼠神经功能损伤程度、HE染色及尼氏染色对大鼠海马DG区、CA3区和CA1区行病理学观察、酶联免疫吸附法测定大鼠海马神经细胞因子酰胆碱酯酶(acetylcholinesterase,AchE)、去甲肾上腺素(norepinephrine,NE)、单胺氧化酶(monoamine oxidase,MAO)水平及测定大鼠海马组织神经元细胞凋亡水平、实时荧光逆转录法及蛋白印记法测定海马组织cAMP、BDNF、NMDA水平。结果与正常对照组比较,砷暴露染毒组、银杏内酯B各剂量组大鼠逃避潜伏期、神经行为评分、海马AchE水平、TUNEL阳性细胞数升高,经过原平台位置的次数、原平台象限停留的时间、海马NE、MAO水平、cAMP(4.34±0.33 vs 1.86±0.15、1.58±0.13 vs 0.25±0.04)、BDNF(4.59±0.43 vs 1.03±0.20、1.38±0.15vs 0.37±0.05)、NMDA(4.87±0.43 vs 1.03±0.11、1.68±0.18 vs 0.33±0.06)mRNA蛋白水平降低,差异均有统计学意义(均P<0.05)。与砷暴露染毒组相比,银杏内酯B各剂量组大鼠逃避潜伏期、神经行为评分、海马AchE水平、TUNEL阳性细胞数降低,经过原平台位置的次数、原平台象限停留的时间、NE、MAO水平、cAMP(1.86±0.15 vs 2.46±0.19、3.54±0.23,0.25±0.04 vs0.78±0.09、1.14±0.13)、BDNF(1.03±0.20 vs 2.34±0.Objective Based on cyclic adenosine monophosphate(cAMP)/brain-derived neurotrophic factor(BDNF)/N-methylD-aspartic acid(N-methyl-D-aspartic acid,NMDA)pathway to explore the protective effect of ginkgolide B on neurological damage in rats exposed to arsenic.Methods Totally 40 SD rats were divided into normal control group,arsenic exposure group(15 mg/kg),low and high dose of ginkgolide B group(200,400 mg/kg).Rats in the arsenic exposure group,low-dose and high-dose ginkgolide B groups were given sodium arsenite by intragastric administration every day for 8 weeks.Ginkgolide B was gavaged,and the normal control group and the arsenic exposure group were given equal volumes of normal saline gavage.After the experiment,the Morris water maze was used to evaluate the spatial learning and memory ability of the rats,and the neurological severity score was used to evaluate the rats'neuropathy and the degree of functional damage.HE staining and Nissl staining was used for pathological observation in rat hippocampus DG area,CA3 area and CA1 area.Enzyme-linked immunosorbent assay was used for the determination of the neurocytokine acetylcholinesterase(AchE)in rat hippocampus,norepinephrine(NE)and monoamine oxidase(MAO)levels,as well as the level of apoptosis in rat hippocampal neurons,and real-time fluorescent reverse transcription and Western blot were used to determine the levels of cAMP,BDNF and NMDA in the hippocampus.Results Compared with the normal control group,the escape latency,neurobehavioral score,hippocampal AchE level,TUNEL positive cell number,times of passing the original platform position,and stay in the original platform quadrant in the arsenic exposure group and the ginkgolide B dose groups were increased.Time,hippocampal NE,MAO levels,cAMP(4.34±0.33 vs1.86±0.15,1.58±0.13 vs 0.25±0.04),BDNF(4.59±0.43 vs 1.03±0.20,1.38±0.15 vs 0.37±0.05),NMDA(4.87±0.43 vs 1.03±0.11,1.68±0.18 vs 0.33±0.06)mRNA protein levels were all decreased,and the differences were statistically significant(all P<0.05).Compared

关 键 词：环磷酸腺苷 脑源性神经营养因子 N-甲基-D-天冬氨酸 银杏内酯B 砷 神经功能损伤 

分 类 号：R-332[医药卫生]