酸枣仁汤对老年慢性快动眼睡眠剥夺模型大鼠心肌线粒体能量代谢的影响  

Effect of Suanzaoren Tang on Energy Metabolism of Myocardial Mitochondria in Aged Rats with Chronic Rapid Eye Movement Sleep Deprivation

作　　者：刘鑫 王平[1] 丁莉[1] 张舜波[1] 石坤 杨忞旻 游秋云[1] LIU Xin;WANG Ping;DING Li;ZHANG Shun-bo;SHI Kun;YANG Min-min;YOU Qiu-yun(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40-46,共7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:从去乙酰化酶沉默信息调节因子3(SIRT3)/超氧化物歧化酶2(SOD2)信号通路研究酸枣仁汤对老年慢性快动眼睡眠剥夺模型大鼠心肌线粒体能量代谢的作用机制。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。采用透射电镜观察心脏线粒体形态,采用比色法检测大鼠下丘脑三磷酸腺苷(ATP)含量的变化,采用分光光度法检测大鼠心肌丙二醛(MDA)含量,SOD活性,采用实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测大鼠心肌SIRT3,SOD2 mRNA和蛋白表达水平,采用免疫荧光染色法检测心肌SIRT3的定位表达。结果:与正常组比较,模型组大鼠心肌组织肌丝排列紊乱,伴有肌丝断裂与溶解,线粒体肿胀明显,线粒体排列紊乱,多数线粒体嵴模糊甚至断裂,心肌ATP含量和SOD活性显著降低(P<0.01),MDA含量显著升高(P<0.01),SIRT3,SOD2 mRNA和蛋白表达显著降低(P<0.01),SIRT3蛋白平均荧光强度显著降低(P<0.01);与模型组比较,酸枣仁汤高剂量组肌丝排列较整齐,线粒体损伤减轻,仅见部分嵴断裂,肿胀程度亦相对较轻,大鼠心肌ATP含量和SOD活性显著升高(P<0.01),MDA含量显著降低(P<0.01),SIRT3,SOD2 mRNA和蛋白表达显著升高(P<0.01),SIRT3蛋白平均荧光强度明显升高(P<0.05),舒乐安定组心肌线粒体损伤也具有一定的改善作用,大鼠心肌SOD活性显著升高(P<0.01),MDA含量显著降低(P<0.01),SIRT3,SOD2 mRNA和蛋白表达显著升高(P<0.01);酸枣仁汤低剂量组心肌线粒体损伤改善作用不明显,但大鼠心肌SOD活性显著升高(P<0.01),SOD2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快动眼睡眠剥夺诱导Objective:To study the mechanism of Suanzaoren Tang in regulating the energy metabolism of myocardial mitochondria in aged rats with chronic rapid eye movement(REM)sleep deprivation through the sirtuin 3(SIRT3)/superoxide dismutase2(SOD2)signaling pathway. 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 myocardial mitochondria was observed under a transmission electron microscope. The content of adenosine triphosphate(ATP)in rat hypothalamus was detected by colorimetry,while the malondialdehyde(MDA)content and the SOD activity in myocardium were measured by spectrophotometry. Real-time polymerase chain reaction(Real-time PCR)and Western blot were conducted to determine the mRNA and protein expression levels of SIRT3 and SOD2 in rat myocardium. The localization of SIRT3 was detected by immunofluorescence staining. Result: Compared with the control group,the model group exhibited a disordered arrangement of myocardial filaments, accompanied by filament rupture and dissolution, obviously swollen mitochondria arranged in disorder,and blurring and even rupture of most mitochondrial cristae. Besides,the content of ATP and SOD activity in the myocardium decreased significantly(P<0.01), whereas that of MDA increased significantly(P<0.01). The mRNA and protein expression levels of SIRT3 and SOD2 were down-regulated significantly(P<0.01),and the average fluorescence intensity of SIRT3 protein declined significantly(P<0.01). The comparison with the model group revealed that high-dose Suanz

关 键 词：酸枣仁汤 睡眠剥夺 透射电镜 能量代谢 

分 类 号：R2-0[医药卫生—中医学] R22