安寐丹调控AMPK信号通路线粒体生物合成对睡眠剥夺大鼠能量代谢紊乱的作用  

Effects of Anmei Pills on the energy metabolism disorders of sleep deprived rats by regulating mitochondrial biosynthesis in the AMPK signaling pathway

作　　者：谢光璟 徐波[1,2] 黄攀攀 王平[1,2] XIE Guang-jing;XU Bo;HUANG Pan-pan;WANG Ping(Hubei University of Traditional Chinese Medicine,Wuhan 430065,China;Institute of Geriatrics,Hubei University of Traditional Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学,武汉430065 [2]湖北中医药大学老年医学研究所,武汉430065

出　　处：《中华中医药杂志》2021年第9期5521-5525,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家重点研发计划(No.2018YFC1705600)。

摘　　要：目的:探讨安寐丹(AMD)对睡眠剥夺大鼠能量代谢的影响。方法:60只大鼠随机分为空白组、模型组、安寐丹组和褪黑素组。通过自制睡眠剥夺箱进行持续性睡眠剥夺4周。力竭性游泳评价活动能力,血生化和酶联免疫吸附实验(ELISA)检测空腹血糖(FBS)、总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)、ATP及ATP酶的含量。HE染色观察下丘脑组织形态,透射电镜观察下丘脑神经元线粒体结构,Western blotting检测下丘脑CaMKKβ、AMPK、PGC-1α蛋白表达,免疫组化和免疫荧光检测Nrf-1、TFAM蛋白表达。结果:与空白组比较,模型组力竭性游泳时间显著降低,FBS、TG、TC、LDL-C水平显著升高,HDL-C水平显著下降,ATP含量、Na-+K-+ATP酶、Ca-2+Mg-2+ATP酶含量显著降低(P<0.01);下丘脑细胞数量减少,胞浆浓缩,核固缩;线粒体结构损坏,嵴断裂,伴有肿胀空泡化;p-CaMKKβ、p-AMPKα、PGC-1α蛋白相对表达显著降低,Nrf-1、TFAM蛋白阳性率显著下降(P<0.01)。与模型组比较,安寐丹组可显著增强大鼠活动耐力,延长游泳时间(P<0.01),降低FBS、TG、TC、LDL-C水平,升高HDL-C水平及ATP、Na-+K-+ATP酶、Ca-2+Mg-2+ATP酶含量(P<0.01),下丘脑细胞数量增加;线粒体水肿减轻,p-CaMKKβ、p-AMPKα、PGC-1α蛋白相对表达升高,Nrf-1、TFAM阳性率增加(P<0.01)。结论:安寐丹改善睡眠剥夺大鼠能量代谢紊乱,其作用可能与促进AMPK通路线粒体生物合成,增加Nrf-1、TFAM蛋白表达有关。Objective:To investigate the effect of Anmei Pills(AMP) on energy metabolism in sleep-deprived rats.Methods:Sixty rats were randomly divided into control group,model group,AMP group and melatonin group.The model was prepared by self-made sleep deprivation box for 4 weeks.Activity was evaluated by exhaustive swimming,and blood chemistry and ELISA were measured hypothalamic blood glucose(FBS),total cholesterol(TC),triglycerides(TG),low-density lipoprotein(LDL-C),high-density lipoprotein(HDL-C),ATP,and ATPase levels.HE staining was for hypothalamic tissue structure,TEM was for mitochondrial structure of hypothalamic neurons,Western blotting was for hypothalamic CaMKK β,AMPK,PGC-1 αprotein expression and immunohistochemistry and immunofluorescence were for Nrf-1,TFAM proteins expression.Results:In the model group,exhaustive swimming time were decreased,FBS,TG,TC,LDL-C levels were increased,and HDL-C,ATP,Na+-K+-ATPase,and Ca2+-Mg2+-ATPase were all decreased(P<0.01);hypothalamic cell number decreased,cytoplasmic concentration,and nuclear consolidation decreased;mitochondrial structure was damaged,cristae ruptured,and swelling and vacuolization;relative expression of p-CaMKK β,p-AMPK,and PGC-1 α proteins were decreased,and Nrf-1 and TFAM protein were positivity decreased(P<0.01).AMP group prolonged swimming time(P<0.01),decreased FB S,TG,TC,LDL-C levels,increased HDL-C and ATP,Na+-K+-ATPase,and Ca2+-Mg2+-ATPase(P<0.01),while alleviating mitochondrial damage,reducing edema vacuolization,and increasing expression of p-CaMKK β,p-AMPK,and PGC-1 α proteins,and Nrf-1 and TFAM are also positivity increasing(P<0.01).Conclusion:AMP ameliorates energy metabolism disorders in sleep-deprived rats,which is probably associating with improving mitochondrial biosynthesis in the AMPK signaling pathway and increase of Nrf-1,TFAM protein expression.

关 键 词：安寐丹 睡眠剥夺 能量代谢 线粒体生物合成 AMP依赖的蛋白激酶(AMPK) 

分 类 号：R285.5[医药卫生—中药学]