电针“足三里”对脾虚大鼠心肌细胞AMPK-线粒体通路的影响  

Effects of electroacupuncture at Zusanli(ST36)on AMPK-mitochondria pathway in cardiac cells of rats with spleen deficiency

作　　者：董佳梓 薛亚楠 魏云涛[3] 张妤[1] 许环宇 张立德[1] Dong Jiazi;Xue Ya’nan;Wei Yuntao;Zhang Yu;Xu Huanyu;Zhang Lide(School of Acupuncture-Moxibustion and Tuiua,Liaoning University of Traditional Chinese Medicine,Liaoning 110847,China;Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences,Beijing 100700;Liaoning Cancer Hospital&Institute,Liaoning 110042,China;Shunde Wu Zhong Pei Hospital,Guandong 528333,China)

机构地区：[1]辽宁中医药大学针灸推拿学院,辽宁110847 [2]中国中医科学院针灸研究所 [3]辽宁省肿瘤医院 [4]广东省佛山市顺德区伍仲珮纪念医院

出　　处：《北京中医药大学学报》2020年第6期522-528,共7页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(No.81603704);辽宁省教育厅一般资助项目(No.L201708)。

摘　　要：目的观察电针“足三里”对脾虚大鼠心肌细胞线粒体能量代谢、线粒体自噬的影响。方法48只SD大鼠,随机分为正常对照组、模型组、足三里组、非经非穴组,每组12只。采用三因素复合法复制脾虚证模型,持续14 d。足三里组针刺大鼠双侧足三里穴,非经非穴组针刺大鼠双侧髂嵴上非经非穴点,电针治疗10 d。高效液相色谱法检测大鼠心肌细胞三磷酸腺苷(ATP)、二磷酸腺苷(ADP)以及磷酸腺苷(AMP)的含量;Western blotting法检测大鼠心肌细胞腺苷酸活化蛋白激酶(AMPK)、磷酸化AMPK(p-AMPK)、微管相关蛋白1轻链3-I(LC3-I)以及LC3-II蛋白的表达;利用透射电镜观察大鼠心肌细胞线粒体及线粒体自噬体的超微结构。结果与正常对照组相比,模型组、足三里组以及非经非穴组大鼠心肌AMP/ATP、LC3-II/LC3-I均明显上调(P﹤0.01);模型组大鼠心肌p-AMPK/AMPK有所上调(P﹤0.05),足三里组大鼠心肌p-AMPK/AMPK显著上调(P﹤0.01)。与模型组相比,足三里组大鼠心肌LC3-II/LC3-I显著上调(P﹤0.01)。模型组大鼠心肌线粒体损伤明显,足三里组线粒体损伤有所改善,见大量的线粒体自噬体。结论电针“足三里”可能通过激活脾虚大鼠心肌细胞AMPK,调节线粒体能量代谢、提高线粒体自噬水平,对脾虚大鼠起到干预作用。Objective To observe the interventional effects of electroacupuncture at Zusanli(ST36)on myocardial mitochondrial energy metabolism and mitophagy of rats with spleen deficiency.Methods Forty-eight SD rats were randomly divided into normal control group,model group,ST36 group and non-acupoint group with twelve rats in each group.Rat models with spleen deficiency were established by means of trifactor method lasting 14 days.The rats in ST36 group were treated with electroacupuncture at Zusanli(ST36)for 10 days,and the rats in non-acupoint group received electroacupuncture at non-acupoint on bilateral iliac crest for 10 days.The contents of myocardial ATP,ADP and AMP of the rats were determined with HPLC.The expressions of myocardial AMPK,p-AMPK and microtubule-associated protein 1 LC3-I and LC3-II of the rats were detected by Western blotting.Ultrastructure of myocardial mitochondria and mitochondrial autophagosome of rats was observed using transmission electron microscope.Results Compared with normal control group,myocardial AMP/ATP and LC3-II/LC3-I ratios of rats in model group,ST36 group and non-acupoint group all significantly increased(P﹤0.01),myocardial p-AMPK/AMPK ratios of rats in model group increased(P﹤0.05)and myocardial p-AMPK/AMPK ratios of rats in ST36 group significantly increased(P﹤0.01).Compared with model group,myocardial LC3-II/LC3-I ratios of rats in ST36 group significantly increased(P﹤0.01).While rats in model group suffered obvious mitochondrial damage,those in ST36 group experienced some improvement with the emergence of a large number of mitochondrial autophagosomes.Conclusion Electroacupuncture at Zusanli(ST36)may be effective in treating rats with spleen deficiency pattern by activating their myocardial AMPK and regulating their myocardial mitochondrial energy metabolism and mitophagy.

关 键 词：足三里 脾虚 心肌细胞 腺苷酸活化蛋白激酶 线粒体 大鼠 

分 类 号：R224.1[医药卫生—中医基础理论]