枸杞多糖对睡眠剥夺大鼠行为能力及激活PI3K/AKT通路的影响  

Effects of lycium barbarum polysaccharide on behavioral ability and PI3K/AKT pathway activation in sleep deprivation rats

作　　者：王艳伟[1] 单铁强[2] 王琮民[3] 姜柳[2] 韩长海 时卫刚 WANG Yanwei;SHAN Tieqiang;WANG Congmin;JIANG Liu;HAN Changhai;SHI Weigang(Department of General Surgery,Handan First Hospital,Handan Hebei,056002,China;Department of Laboratory,Qinhuangdao Haigang Hospital,Qinhuangdao Hebei,066000,China;Medical College,Hebei University of Engineering,Handan Hebei,056002,China;Department of Anesthesiology,Handan First Hospital,Handan Hebei,056002,China)

机构地区：[1]邯郸市第一医院综合外科,河北邯郸056002 [2]秦皇岛市海港医院检验科,河北秦皇岛066000 [3]河北工程大学医学院,河北邯郸056002 [4]邯郸市第一医院麻醉科,河北邯郸056002

出　　处：《职业与健康》2024年第8期1041-1045,共5页Occupation and Health

基　　金：河北省中医药管理局科研计划项目(2023429)。

摘　　要：目的 分析枸杞多糖对睡眠剥夺大鼠海马区自噬蛋白、磷脂酰肌醇3激酶(phosphatidylinositol 3 kinase,PI3K)/丝苏氨酸激酶(serine threonine kinase,AKT)信号通路以及大鼠行为能力的影响,探讨枸杞多糖改善大鼠睡眠剥夺的分子机制。方法 利用“多平台水环境”建立实验性剥夺睡眠大鼠18只将其分为睡眠剥夺组、枸杞多糖组和艾司唑仑3个组,选择健康大鼠为正常组。枸杞多糖组和艾司唑仑组每日分别灌胃枸杞多糖和艾司唑仑,睡眠剥夺组和正常组每日灌胃相同体积的三蒸水,4个组均持续灌胃28 d。通过“水迷宫”实验检测大鼠行为能力变化;酶联免疫吸附法检测海马组织中丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)水平;免疫印迹技术评估大鼠海马区自噬蛋白:Bcl-2同源结构域蛋白抗体(bcl-2 homologous domain protein antibody,Beclin 1)及微管相关蛋白轻链3Ⅱ(microtubule-associated protein light chain 3Ⅱ,LC3Ⅱ)/微管相关蛋白轻链3Ⅰ(microtubule-associated protein light chain 3Ⅰ,LC3Ⅰ)的含量;PI3K及磷酸化磷酸肌醇3激酶(phosphorylated inositol 3 kinase,p-PI3K)、AKT及磷酸化丝苏氨酸激酶(phosphorylated serine threonine kinase,p-AKT)的含量。结果 睡眠剥夺组与正常组相比,大鼠的逃避潜伏期[(41.98±5.27)s]、延长穿台次数[(9.56±1.46)次]减少(均P<0.05);海马组织中MDA[(3.55±0.46)mmol/L]含量升高、SOD[(7.62±1.20)U/L]含量降低(均P<0.01);海马区自噬蛋白Beclin 1(1.60±0.17)、LC3Ⅱ/LC3Ⅰ(1.18±0.14)的含量均升高(均P<0.05),p-PI3K/PI3K(0.28±0.05)及p-AKT/AKT(0.40±0.07)含量均降低(均P<0.05)。枸杞多糖组与睡眠剥夺组相比,大鼠逃避潜伏期[(29.79±4.01)s]缩短、穿台次数[(16.01±2.29)次]增多(均P<0.05);海马组织中MDA[(2.31±0.38)mmol/L]含量降低、SOD[(14.25±1.89)U/L]含量升高(均P<0.01);海马区自噬蛋白Beclin 1(1.02±0.16)、LC3Ⅱ/LC3Ⅰ(0.80±0.12)的含量均降低(均P<0.05),p-PIObjective To analyze the effects of lycium barbarum polysaccharide on autophagy protein,phosphatidylinositol 3 kinase(PI3K)/threonine protein kinase(AKT) signaling pathway and behavioral ability in hippocampus of sleep deprivation rats,and explore the molecular mechanism of lycium barbarum polysaccharide improving sleep deprivation rats.Methods The sleep deprived experimental was established in rats by using the "multiplatform water environment",18 rats were arranged into three groups:sleep deprivation group,lycium barbarum polysaccharide group and estazolam group.The health rats were arranged into the normal group.The lycium barbarum polysaccharide group and estazolam group were given daily gavage with lycium barbarum polysaccharide and estazolam respectively,sleep deprivation group and normal groupwere given daily gavage with the same volume of triple distilled water.All the four groups were continuously fed for 28 days.The changes of rats' behavioral abilities was measured through the "water maze" experiment.The enzyme-linked immunosorbent assay was utilized to measure the level of malondialdehyde(MDA) and superoxide dismutase(SOD) in hippocampus.The Western blot was utilized to determine the levels of bcl-2 homologous domain protein antibody(Beclin 1),microtubule-associated protein light chain 3Ⅱ (LC3Ⅱ)/microtubuleassociated protein light chain 3Ⅰ(LC3Ⅰ),PI3K,phosphorylated inositol 3 kinase(p-PI3K),Akt and phosphorylated serine threonine kinase(p-AKT) in the hippocampus of rats.Results Compared with the normal group,the rats' escape latency[(41.98±5.27)s] in the sleep deprivation group was prolonged,and the number of crossing platforms[(9.56±1.46)times] was reduced(both P<0.05),the content of MDA[(3.55±0.46)mmol/L] increased and the content of SOD[(7.62±1.20)U/L] decreased in the hippocampus tissue(both P<0.01),the contents of autophagic proteins Beclin 1(1.60±0.17) and LC3Ⅱ/LC3Ⅰ(1.18±0.14) in the hippocampus increased(both P<0.05),while the contents of p-PI3K/PI3K(0.28±0.05) and p-AKT/AKT(

关 键 词：枸杞多糖 睡眠剥夺 自噬蛋白 PI3K/AKT通路 行为能力 

分 类 号：R114[医药卫生—卫生毒理学]