酸枣仁汤对老年慢性快动眼睡眠剥夺模型大鼠肝脏线粒体能量代谢及其机制的影响  

Effect of Suanzaoren Tang on Energy Metabolism of Liver Mitochondria in Aged Rats with Chronic Rapid Eye Movement Sleep Deprivation and Its Mechanism

作　　者：刘鑫 游秋云[1] 王平[1] 丁莉[1] 宁富楠 张舜波[1] LIU Xin;YOU Qiu-yun;WANG Ping;DING Li;NING Fu-nan;ZHANG Shun-bo(Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学,武汉430065

出　　处：《中国实验方剂学杂志》2021年第16期53-58,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81473560)。

摘　　要：目的:研究酸枣仁汤对老年慢性睡眠剥夺模型大鼠肝脏线粒体能量代谢的影响及作用机制。方法:将50只雄性Wistar大鼠随机分为正常组,模型组、舒乐安定组(0.18 mg·kg^(-1)·d^(-1)),酸枣仁汤低、高剂量组(6.48,12.96 g·kg^(-1)·d^(-1)),除正常组外,其余各组均皮下注射D-半乳糖,于末次给药后进行多平台水环境睡眠剥夺,造模结束后各组开始灌胃给药,连续给药7 d。采用透射电镜观察肝脏线粒体形态,采用比色法检测大鼠肝脏三磷酸腺苷(ATP)含量的变化,采用比色法检测大鼠肝脏线粒体呼吸链还原型辅酶Ⅰ(复合物Ⅰ),琥珀酸氧化还原酶(复合物Ⅱ),细胞色素C氧化还原酶(复合物Ⅲ),细胞色素C氧化酶(复合物Ⅳ)活性,采用蛋白免疫印迹法(Western blot)检测大鼠肝脏柠檬酸合酶(CS),异柠檬酸脱氢酶(IDH),负责编码ATP合成酶F0亚基的一段核基因(ATP5F1)蛋白的表达水平。结果:与正常组比较,模型组大鼠肝脏内线粒体损伤较严重,线粒体发生肿胀、变形,嵴断裂并且数量有一定的减少,与模型组比较,舒乐安定组、酸枣仁汤高剂量组线粒体损伤减轻,肿胀、变形情况显著减少,嵴断裂减少,其中酸枣仁汤高剂量改善较为明显。与正常组比较,模型组大鼠肝脏ATP含量显著降低(P<0.01),线粒体呼吸链复合物Ⅰ,Ⅱ,Ⅲ,Ⅳ活性均显著降低(P<0.01),IDH,CS,ATP5F1蛋白表达均显著降低(P<0.01);与模型组比较,酸枣仁汤高剂量组肝脏ATP含量显著升高(P<0.01),线粒体呼吸链复合物Ⅰ,Ⅱ,Ⅲ,Ⅳ活性均明显升高(P<0.05,P<0.01),IDH,CS,ATP5F1蛋白表达均显著升高(P<0.01),舒乐安定组肝脏ATP含量明显升高(P<0.05),线粒体呼吸链复合物Ⅰ,Ⅲ活性显著升高(P<0.01),CS,ATP5F1蛋白表达均显著升高(P<0.01),酸枣仁汤低剂量组肝脏线粒体呼吸链复合物Ⅰ,Ⅲ活性明显升高(P<0.05,P<0.01),ATP5F1蛋白表达明显升高(P<0.05)。结论:酸枣仁汤可以改善老年慢性睡眠剥夺引Objective: To study the effect of Suanzaoren Tang on energy metabolism of liver mitochondria in aged rats with chronic rapid eye movement(REM)sleep deprivation. Method: Fifty male Wistar rats were randomly divided into the control group,model group,estazolam group(0.18 mg·kg^(-1)·d^(-1)),and low-(6.48 g·kg^(-1)·d^(-1))and high-dose(12.96 g·kg^(-1)·d^(-1))Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of D-galactose and then were deprived of sleep using the multiple platform method after the last administration. Following successful modeling,the rats in each group were treated with intragastric administration of the corresponding drugs for seven consecutive days. The morphology of liver mitochondria was observed under the transmission electron microscope. The content of adenosine triphosphate(ATP) in rat liver was detected by colorimetry, and the activities of ubiquinone oxidoreductase(complex Ⅰ),succinate-ubiquinone oxidoreductase(complex Ⅱ),ubiquinol-cytochrome c oxidoreductase(complex Ⅲ),and cytochrome c oxidase(complex Ⅳ)in mitochondrial respiratory chain of rat liver were measured by colorimetry. The protein expression levels of citrate synthase(CS), isocitrate dehydrogenase(IDH),and ATP synthase,H+transporting,mitochondrial F0 complex,subunit b,isoform 1(ATP5 F1)in rat liver were assayed by Western blot. Result:The mitochondrial damage in rat liver of the model group was more serious than that in the control group,manifested as mitochondrial swelling and deformation as well as cristae rupture and reduction. The comparison with the model group revealed that both the positive control and Suanzaoren Tang at the high dose obviously alleviated the mitochondrial swelling and deformation and reduced cristae rupture,with better improvements observed in the high-dose Suanzaoren Tang group. Compared with the control group,the content of ATP,the activities of mitochondrial respiratory chain complexes Ⅰ,Ⅱ,Ⅲ,and Ⅳ,and the protein expression le

关 键 词：酸枣仁汤 睡眠剥夺 透射电镜 肝脏 能量代谢 

分 类 号：R2-0[医药卫生—中医学] R22