摘要
目的研究葛根异黄酮对去卵巢大鼠心肌损伤的保护作用及其机制。方法将36只大鼠随机分为假手术组、模型组、戊酸雌二醇组(0.1 mg/kg)和葛根异黄酮低、中、高剂量组(55、110、220 mg/kg)。模型组和各给药组大鼠切除双侧卵巢,假手术组大鼠仅切除卵巢周边小块脂肪组织。手术后2周开始给药,每天1次,每周6 d,连续灌胃给药16周。第16周末,通过PowerLab检测血流动力学[收缩压(SBP)、舒张压(DBP)、平均压(MBP)、左室收缩压(LVSP)、左室舒张压(LVMP)、等容收缩期左心室内压力最大上升和下降速率(±dp/dt_(max))];HE染色观察心脏组织病理变化;生化分析仪检测血浆中肌酸激酶(CK)、乳酸脱氢酶(LDH)、总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)及葡萄糖(Glu)水平;比色法检测心肌组织中三磷酸腺苷(ATP)水平;RT-qPCR法检测心肌组织中葡萄糖转运蛋白4(GLUT 4)、乳酸脱氢酶A(LDHA)、肉毒碱棕榈酰基转移酶-1(CPT-1α)、酰基辅酶A-羧化酶(ACC)、肝激酶B1(LKB 1)、腺苷酸活化蛋白激酶(AMPK)、过氧化物酶体增殖物激活受体γ辅激活酶因子1α(PGC-1α)mRNA表达;Western blot法检测心肌组织中能量代谢相关蛋白LKB1、p-AMPK/AMPK、PGC-1α表达。结果与模型组比较,葛根异黄酮组SBP、DBP、MBP、LVSP、LVMP降低(P<0.05,P<0.01),-dp/dt_(max)升高(P<0.05,P<0.01);去卵巢所致大鼠心肌纤维溶解、间隙增宽及炎性浸润情况得到改善;LDH、CK活性降低(P<0.05);心肌组织ATP水平升高(P<0.05,P<0.01);TC、TG、LDL-C及Glu水平降低(P<0.05,P<0.01),HDL-C水平升高(P<0.05,P<0.01);心肌组织GLUT4、LDHA、CPT-1α、ACC、LKB1、AMPK、PGC-1αmRNA表达升高(P<0.05,P<0.01);心肌组织LKB1、p-AMPK/AMPK、PGC-1α蛋白表达升高(P<0.05,P<0.01)。结论葛根异黄酮对去卵巢所致的大鼠心肌损伤具有保护作用,其机制与改善心肌组织能量代谢相关蛋白,调控LKB1/AMPK/PGC-1α能量代谢相关信号通路有关。
AIM To study the protective effects and mechanism of pueraria isoflavones on myocardial injury in ovariectomized rats.METHODS Thirty-six rats were randomly divided into the sham operation group,the model group,the estradiol valerate group(0.1 mg/kg)and the low,medium and high dose pueraria isoflavones groups(55,110,220 mg/kg).In contrast to the rats of the sham operation group having their small pieces of adipose tissue removal around the ovaries,rats of the other groups had their bilateral ovaries excised,followed by the 16-week corresponding oral drug administration 2 weeks later at a once daily frequency for,6 days a week.At the end of the 16th week,the rats had their hemodynamics[systolic pressure(SBP),diastolic pressure(DBP),mean pressure(MBP),left ventricular systolic pressure(LVSP),left ventricular diastolic pressure(LVMP),and the maximum rate of increase and decrease of left ventricular pressure during isovolumic contraction(±dp/dt_(max))]detected by PowerLab;their cardiac pathological changes observed by HE staining;their levels of creatine kinase(CK),lactate dehydrogenase(LDH),total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C)and glucose(Glu)in plasma detected by biochemical analyzer;their myocardial level of adenosine triphosphate(ATP)detected by colorimetry;their mRNA expressions of glucose transporter 4(GLUT 4),lactate dehydrogenase A(LDHA),carnitine palmitoyl transferase-1(CPT-1α),acyl coenzyme A carboxylase(ACC),liver kinase B1(LKB 1),adenylate-activated protein kinase(AMPK)and peroxisome proliferator-activated receptorγcoactivator factor 1α(PGC-1α)detected by RT-qPCR;and their myocardial expressions of energy metabolism related proteins LKB1,p-AMPK/AMPK and PGC-1αdetected by Western blot.RESULTS Compared with the model group,the pueraria isoflavones groups displayed decreased levels of SBP,DBP,MBP,LVSP,LVMP(P<0.05,P<0.01);increased-dp/dt_(max)(P<0.05,P<0.01);improved myocardial fibrinolysis,gap widening and inflammatory infiltration caused by ovariectomy;decreased activities of LDH and CK(P<0.05);increased myocardial ATP level(P<0.05,P<0.01);decreased levels of TC,TG,LDL-C and Glu(P<0.05,P<0.01);increased HDL-C level(P<0.05,P<0.01);increased myocardial mRNA expressions of GLUT4,LDHA,CPT-1α,ACC,LKB1,AMPK and PGC-1α(P<0.05,P<0.01);and increased protein expressions of myocardial LKB1,p-AMPK/AMPK and PGC-1α(P<0.05,P<0.01).CONCLUSION Pueraria isoflavones are protective to myocardial injury in ovariectomized rats,and the mechanism may lie in the improvement of energy metabolism-related myocardial proteins via LKB1/AMPK/PGC-1αsignaling pathway.
作者
张莹
欧阳灿玥
陈兰英
袁蓓欣
崔宏伟
谢欣序
刘鹏
刘荣华
ZHANG Ying;OUYANG Can-yue;CHEN Lan-ying;YUAN Bei-xin;CUI Hong-wei;XIE Xin-xu;LIU Peng;LIU Rong-hua(Jiangxi University of Chinese Medicine,Nanchang 330004,China;National Pharmaceutical Engineering Centre for Solid Preparations of Chinese Herbal Medicine,Nanchang 330006,China)
出处
《中成药》
CAS
CSCD
北大核心
2024年第8期2542-2551,共10页
Chinese Traditional Patent Medicine
基金
国家重点研发计划项目(2017YFC1702902)。
作者简介
张莹(1998-),女,硕士生,从事中药药效评价与作用机制研究。Tel:13820232057,E-mail:3209598566@qq.com;通信作者:陈兰英(1966-),女,博士,教授,从事中药药效评价与作用机制研究。Tel:(0791)87119611,E-mail:clyxy2513@163.com;通信作者:刘荣华(1964-),男,博士,教授,从事中药药效物质基础与中药质量评价研究。Tel:(0791)87118992,E-mail:rhliu@163.com。
