摘要
目的探讨基于核因子E2相关因子2(NRF2)/膜铁转运蛋白1(FPN1)通路对黄芪甲苷(AS)减轻糖尿病(DM)心肌梗死大鼠心肌损伤的作用机制。方法通过高糖饲料及链脲佐菌素(STZ)建立DM大鼠模型,将DM造模成功的50只大鼠通过结扎左冠状动脉前降支构建DM心肌梗死大鼠模型,将造模成功的50只大鼠随机分为梗死组、AS低剂量(AS-L,20 mg/kg AS)组、AS中剂量(AS-M,40 mg/kg AS)组、AS高剂量(AS-H,80 mg/kg AS)组、AS-H+NRF2抑制剂(ML385,80 mg/kg AS+30 mg/kg ML385)组,同时以10只开胸大鼠设为假手术组。干预结束后,检测心功能[左心室射血分数(LVEF)、肌酸激酶同工酶-MB(CK-MB)、肌酸激酶(CK)]指标;2,3,5-氯化三苯基四唑(TTC)检测心肌梗死体积比;试剂盒检测铁含量水平;HE染色检测大鼠心肌组织病理变化;qRT-PCR及Western blot分析心肌组织中NRF2、谷胱甘肽过氧化酶4(GPX4)、FPN1mRNA及蛋白表达。结果与假手术组相比,梗死组大鼠病理损伤严重,LVEF、NRF2、GPX4、FPN1mRNA及蛋白表达显著降低,梗死体积比、铁离子含量、CK-MB、CK显著增加(均P<0.05);与梗死组相比,AS-L组、AS-M组、AS-H组大鼠病理损伤得到改善,LVEF、NRF2、GPX4、FPN1mRNA及蛋白表达显著增加,梗死体积比、铁离子含量、CK-MB、CK显著降低,呈现剂量依赖性(均P<0.05);与AS-H组相比,AS-H+ML385组病理损伤相对严重,LVEF、NRF2、GPX4、FPN1mRNA及蛋白表达显著降低,梗死体积比、铁离子含量、CK-MB、CK显著增加(均P<0.05)。结论AS可减轻DM心肌梗死大鼠心肌损伤,可能通过激活NRF2/FPN1通路抑制铁死亡实现。
Objective To explore the mechanism of astragaloside A(AS)in alleviating myocardial injury in rats with myocardial infarction in diabetes mellitus(DM)based on nuclear factor E2-related factor 2(NRF2)/ferroportin 1(FPN1)pathway.Methods DM rat model was established by high sugar diet and streptozotocin(STZ).The 50 rats successfully modeling DM was giving ligating the anterior descending branch of left coronary artery to establish the rat model of DM myocardial infarction.The 50 rats with successful modeling were randomly divided into infarction group,low dose AS(AS-L,20 mg/kg AS),middle dose AS(AS-M,40 mg/kg AS),high dose AS(AS-H,80 mg/kg AS)group,and AS-H+NRF2 inhibitor(ML385,80 mg/kg AS+30 mg/kg ML385)group.Meantime,10 open chest rats were taken as the sham operation group.After the intervention,cardiac function[left ventricular ejection fraction(LVEF),creatine kinase isoenzyme-MB(CK-MB),creatine kinase(CK)]indexes were measured;2,3,5-triphenyltetrazolium chloride(TTC)was used to measure the volume ratio of myocardial infarction.The kit was used to detect the iron content level;HE staining was used to detect the pathological changes of myocardium in rats.mRNA and proten expression of NRF2,glutathione peroxidase 4(GPX4)and FPN1 in myocardium eve analyzed by qRT-PCR and Western blot.Results Compared with the sham operation group,the pathological injury of rats in the infarction group was serious,the expression of LVEF,mRNA and protein expression of NRF2,GPX4 and FPN1 decreased obviously,the infarct volume ratio,iron ion content,CK-MB and CK increased obviously(P<0.05).Compared with the infarction group,the pathological injury of rats in AS-L group,AS-M group and AS-H group was improved,the expression of LVEF,mRNA and protein expression of NRF2,GPX4 and FPN1 increased obviously,the infarct volume ratio,iron ion content,CK-MB and CK decreased obviously,they were dose dependent(P<0.05).Compared with the AS-H group,the pathological injury in the AS-H+ML385 group was relatively serious,the expression of LVEF,mRNA and pro
作者
邓长青
张京兰
金海涛
DENG Changqing;ZHANG Jinglan;JIN Haitao(Department of Encephalopathy,Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430014,China)
出处
《西部医学》
2023年第10期1439-1443,1451,共6页
Medical Journal of West China
基金
武汉市医学科研项目(WZ20C36)。
