基于AMPK/mTOR通路探讨电针“丰隆”穴对高脂血症大鼠肝脏胆固醇代谢的影响  

Effect of electroacupuncture at"Fenglong"(ST 40)on liver cholesterol metabolism in hyperlipidemia rats based on AMPK/mTOR pathway

作　　者：金舒文 刘嘉宝 李丹 张茜 刘曼琦 张红星 潘小丽 JIN Shuwen;LIU Jiabao;LI Dan;ZHANG Xi;LIU Manqi;ZHANG Hongxing;PAN Xiaoli(College of Acupuncture-Moxibustion and Orthopaedics,Hubei University of CM,Wuhan 430061,China;Medical Department,Jianghan University;Acupuncture and Moxibustion Research Institute of Jianghan University,Wuhan 430056,Hubei Province;Hubei Shizhen Laboratory,Wuhan 430061)

机构地区：[1]湖北中医药大学针灸骨伤学院,武汉430061 [2]江汉大学医学部 [3]江汉大学针灸研究所,湖北武汉430056 [4]湖北时珍实验室,武汉430061

出　　处：《中国针灸》2024年第10期1155-1164,共10页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金资助项目:82274663;湖北省教育厅科学研究计划资助项目:Q20212008。

摘　　要：目的:观察电针“丰隆”穴对高脂血症(HPL)大鼠腺苷酸活化蛋白激酶(AMPK)、哺乳动物雷帕霉素靶蛋白(mTOR)表达水平及下游胆固醇代谢相关分子胆固醇调节元件结合蛋白-2(SREBP-2)、3-羟基3-甲基戊二酰辅酶A还原酶(HMGCR)、三磷酸腺苷结合盒转运体G5/G8(ABCG5/G8)表达的影响,探讨电针干预HPL的可能机制。方法:将30只SPF级雄性SD大鼠随机分为空白组、模型组、AMPK激动剂组、电针组和电针+AMPK抑制剂组,每组6只。采用高脂饲料喂养法制备HPL模型。造模完成后,电针组大鼠行电针干预,穴取双侧“丰隆”,予疏密波,频率2 Hz/100 Hz,电流1 mA,每次30 min,每天1次;AMPK激动剂组予以腹腔注射AMPK激动剂A-769662(30 mg/kg),每天2次;电针+AMPK抑制剂组于电针干预前30 min腹腔注射AMPK抑制剂化合物C(Compound C,25 mg/kg),每天1次。3组均每周连续干预5 d,干预4周。全自动生化分析仪检测大鼠血脂相关指标[血清胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)及丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)含量];HE染色和油红O染色法观察大鼠肝脏组织形态;称重计算肝指数,ELISA法检测大鼠肝脏组织TC、TG含量及粪便TC、胆汁酸含量;Western blot法检测大鼠肝脏组织AMPK、m TOR蛋白磷酸化水平;免疫组化法检测大鼠肝脏组织SREBP-2、HMGCR和ACBG5/G8阳性表达。结果:造模后,模型组、AMPK激动剂组、电针组和电针+AMPK抑制剂组大鼠血清TC、TG和LDL-C含量均高于空白组(P<0.01);各组大鼠血清HDL-C含量比较,差异无统计学意义(P>0.05)。干预后,与空白组比较,模型组大鼠血清TC、TG、LDL-C、ALT、AST含量,肝指数,肝脏组织TC、TG含量及粪便TC、胆汁酸含量均升高(P<0.01);HE和油红O染色可见肝细胞排列紊乱,细胞内有明显的大泡性脂滴,明显脂质蓄积;肝脏组织磷酸化腺苷酸活化蛋白激酶(p-AMPK)蛋白表达、p-AObjective To observe the effects of electroacupuncture(EA)at"Fenglong"(ST 40)on the expression of adenosine 5'-monophosphate-activated protein kinase(AMPK),mammalian target of rapamycin(mTOR),and the expression of the downstream molecules related to cholesterol metabolism i.e.sterol regulatory element binding protein-2(SREBP-2),recombinant 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMGCR),and adenosine triphosphate binding cassette transporter G5/G8(ABCG5/G8)in the rats with hyperlipidemia(HPL)so as to explore the possible mechanism of EA in the intervention of HPL.Methods Thirty SPF-grade male SD rats were randomly divided into a blank group,a model group,an AMPK agonist group,an EA group,and an EA+AMPK inhibitor group,6 rats in each group.The high-fat feeding method was adopted to establish HPL model.After successfully modeled,the rats in the EA group received EA intervention at bilateral"Fenglong"(ST 40),with disperse-dense wave,in the frequency of 2 Hz/100 Hz,the intensity of 1 mA.EA was given once daily,for 30 min in one intervention.In the AMPK agonist group,the intraperitoneal injection with AMPK agonist A-769662was administered,30 mg/kg,twice a day.In the EA+AMPK inhibitor group,the intraperitoneal injection of AMPK inhibitor Compound C was administered,25 mg/kg,once a day,30 min before EA intervention.In the intervention groups,the interventions were delivered continuously for 5 days a week and lasted 4 weeks.Using automated biochemical analyzer,the blood lipid-related indexes(serum total cholesterol[TC],triglycerides[TG],low-density lipoprotein cholesterol[LDL-C]and high-density lipoprotein cholesterol[HDL-C]as well as alanine aminotransferase[ALT]and aspartate aminotransferase[AST])were detected in the rats.HE staining and oil red O staining were used to observe the morphology of liver tissue.Liver index was calculated by the weight.Using ELISA,the contents of TC and TG of liver tissue and the contents of of TC and bile acid in feces were detected.The protein phosphorylation levels of AMPK and mTO

关 键 词：高脂血症 电针 丰隆 胆固醇代谢 腺苷酸活化蛋白激酶/哺乳动物雷帕霉素靶蛋白 

分 类 号：R245[医药卫生—针灸推拿学]