首页|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

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

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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-asso-ciated 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-PI3K/PI3K(0.52± 0.08)及p-AKT/AKT(0.64±0.09)含量均升高(均P<0.05).枸杞多糖组和艾司唑仑组相比,上述指标差异均无统计学意义(均P>0.05).结论 枸杞多糖可以通过激活PI3K/AKT通路,抑制海马的过度自噬来改善睡眠剥夺大鼠的行为能力.
Effects of lycium barbarum polysaccharide on behavioral ability and PI3K/AKT pathway activation in sleep deprivation rats
Objective 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 3 Ⅱ(LC3 Ⅱ)/microtubule-associated protein light chain 3 Ⅰ(LC3 Ⅰ),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 LC3 Ⅱ/LC3 Ⅰ(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(0.40±0.07)decreased(both P<0.05).Compared with the sleep deprivation group,the rats'escape latency[(29.79±4.01)s]in the ycium barbarum polysaccharide group was is shortened,and the number of crossing platforms[(16.01±2.29)times]was increased(both P<0.05),the content of MDA[(2.31±0.38)mmol/L]decreased and the content of SOD[(14.25±1.89)U/L]increased in the hippocampus tissue(both P<0.01),the contents of autophagic proteins Beclin 1(1.02±0.16)and LC3 Ⅱ/LC3 Ⅰ(0.80±0.12)in the hippocampus decreased(both P<0.05),while the contents of p-PI3K/PI3K(0.52±0.08)and p-AKT/AKT(0.64±0.09)increased(both P<0.05).The lycium barbarum polysaccharide group and estazolam group had not obvious difference in the above indicators(both P>0.05).Conclusion The lycium barbarum polysaccharide is able to upgrade the behavioral ability of sleep deprivation rats by activating PI3K/Akt pathway and inhibiting excessive autophagy in hippocampus.

Lycium barbarum polysaccharidesSleep deprivationAutophagy proteinPI3K/AKT pathwayBehavioral ability

王艳伟、单铁强、王琮民、姜柳、韩长海、时卫刚

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邯郸市第一医院综合外科,河北邯郸 056002

秦皇岛市海港医院检验科,河北秦皇岛 066000

河北工程大学医学院,河北邯郸 056002

邯郸市第一医院麻醉科,河北邯郸 056002

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枸杞多糖 睡眠剥夺 自噬蛋白 PI3K/AKT通路 行为能力

河北省中医药局科研项目

2023429

2024

职业与健康
天津市疾病预防控制中心 中华预防医学会

职业与健康

CSTPCD
影响因子:0.737
ISSN:1004-1257
年,卷(期):2024.40(8)
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