参麦注射液调控铜死亡在心肌纤维化大鼠中的作用及机制研究  

Effect and mechanism of Shenmai Injection in regulating copper death in myocardial fibrosis in rats

作　　者：刘思彤 郭志远 邹樾 谌治安 张帅 王艳[1] 王丽英 张艺红[1] 刘智[1] LIU Si-tong;GUO Zhi-yuan;ZOU Yue;CHEN Zhi-an;ZHANG Shuai;WANG Yan;WANG Li-ying;ZHANG Yi-hong;LIU Zhi(Changchun University of Chinese Medicine,Changchun 130117,China;the Second Norman Bethune Hospital of Jilin University,Changchun 130041,China)

机构地区：[1]长春中医药大学,吉林长春130117 [2]吉林大学白求恩第二医院,吉林长春130041

出　　处：《中国中药杂志》2025年第6期1601-1609,共9页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81603324);吉林省科技发展计划面上项目(YDZJ202401052ZYTS);吉林省大学生创新训练计划项目(S202210199041)。

摘　　要：基于铜死亡探讨参麦注射液对异丙肾上腺素(isoproterenol,ISO)所诱导大鼠心肌纤维化(myocardiac fibrosis,MF)的影响及作用机制。将SPF级雄性SD大鼠随机分为正常组、模型组、卡托普利(5 mg·kg^(-1))阳性对照组及参麦注射液低(6 mL·kg^(-1))、中(9 mL·kg^(-1))、高(12 mL·kg^(-1))剂量组。除正常组外,其余各组大鼠均采用皮下注射ISO(5 mg·kg^(-1)),每日1次,连续10 d的方法建立大鼠MF模型,造模成功后第2天开始腹腔注射给药,连续28 d。苏木素-伊红(hematoxylin-eosin,HE)、Masson染色观察大鼠心肌组织病理学变化及纤维化水平;比色法检测大鼠血清Cu^(2+)浓度;酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测大鼠血清中炎性因子白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-18(interleukin-18,IL-18)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平以及线粒体能量代谢三磷酸腺苷(adenosine triphosphate,ATP)、二磷酸腺苷(adenosine diphosphate,ADP)、磷酸腺苷(adenosine monophosphate,AMP)水平;Wes-tern blot检测大鼠心肌组织Ⅰ型胶原(collagen Ⅰ,Col-Ⅰ)、Ⅲ型胶原(collagen Ⅲ,Col-Ⅲ)和铜死亡相关蛋白二氢硫辛酸乙酰转移酶(dihydrolipoamide acetyltransferase,DLAT)、铁氧还蛋白1(ferredoxin 1,FDX1)、硫辛酸合酶(lipoic acid synthase,LIAS)、热休克蛋白70(heat shock protein 70,HSP70)的表达;免疫荧光检测DLAT、FDX1、HSP70的表达;免疫组化检测DLAT、FDX1、LIAS、HSP70的表达。结果显示,与模型组比较,各给药组大鼠心肌结构紊乱和胶原纤维沉积明显改善,心脏指数水平降低,血清Cu^(2+)浓度、IL-6、IL-1β、IL-18、TNF-α及ADP、AMP水平显著降低,ATP水平显著升高,心肌组织中Col-Ⅰ、Col-Ⅲ及HSP70蛋白表达显著降低,DLAT、FDX1、LIAS蛋白表达显著升高。综上所述,参麦注射液能有效缓解ISO诱导的MF大鼠心肌结构紊乱和间质胶原纤维化沉积,可�Based on copper death,this study investigates the effect and mechanism of Shenmai Injection on isoproterenol(ISO)-induced myocardial fibrosis(MF)in rats.SPF-grade male SD rats were randomly divided into a normal group,model group,captopril(5 mg·kg^(-1))positive control group,and Shenmai Injection low(6 mL·kg^(-1)),medium(9 mL·kg^(-1)),and high(12 mL·kg^(-1))dose groups.Except for the normal group,the rats in the other groups were subcutaneously injected with ISO(5 mg·kg^(-1))once a day for 10 consecutive days to establish an MF model.Starting from the second day after successful modeling,intraperitoneal injections of the respective treatments were administered for 28 consecutive days.Hematoxylin-eosin(HE)and Masson staining were used to observe pathological changes and fibrosis levels in the myocardial tissue.Colorimetry was employed to detect serum Cu^(2+)concentration in rats.The levels of inflammatory cytokines interleukin-6(IL-6),interleukin-1β(IL-1β),interleukin-18(IL-18),tumor necrosis factor-α(TNF-α),as well as mitochondrial energy metabolites adenosine triphosphate(ATP),adenosine diphosphate(ADP),and adenosine monophosphate(AMP)in serum were measured using enzyme-linked immunosorbent assay(ELISA).Western blot was performed to detect the expression of collagen Ⅰ(Col-Ⅰ),collagen Ⅲ(Col-Ⅲ),and copper death-related proteins dihydrolipoamide acetyltransferase(DLAT),ferredoxin 1(FDX1),lipoic acid synthetase(LIAS),and heat shock protein 70(HSP70)in myocardial tissue.Immunofluorescence was used to detect the expression of DLAT,FDX1,and HSP70,while immunohistochemistry was conducted to examine the expressions of DLAT,FDX1,LIAS,and HSP70.The results showed that,compared to the model group,the myocardial structure disorder and collagen fiber deposition in the drug treatment groups were significantly improved,the cardiac index level was reduced,serum Cu^(2+),IL-6,IL-1β,IL-18,TNF-α,ADP,and AMP levels were significantly decreased,ATP levels were significantly increased,and the expressions of Col-Ⅰ,C

关 键 词：参麦注射液 心肌纤维化 铜死亡 二氢硫辛酸乙酰转移酶(DLAT) 

分 类 号：R285.5[医药卫生—中药学]