暖心康通过“代谢-炎症”网络调控巨噬细胞极化对心肌梗死后小鼠心室重构的影响  

Pharmacological Effect of Nuanxinkang Powder on Ventricular Remodeling in Post-infarction Mice Through"Metabolic-Inflammatory"Network Regulating Macrophage Polarization

作　　者：林祉均 陈梓欣[1] 江佳林 董鑫 关卓骥 王陵军[1] LIN Zhijun;CHEN Zixin;JIANG Jialin;DONG Xin;GUAN Zhuoji;WANG Lingjun(The First Affiliated Hospital of Guangzhou University of Chinese Medicine/National Key Laboratory of Traditional Chinese Medicine Syndrome,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学第一附属医院/中医证候全国重点实验室,广东广州510405

出　　处：《中药新药与临床药理》2024年第2期159-167,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金青年科学基金项目(82004293);广州市科技计划重点研发计划项目(202206080015)。

摘　　要：目的探究暖心康(红参、毛冬青)通过“代谢-炎症”网络调控巨噬细胞极化改善心肌梗死后小鼠心室重构的作用机制。方法(1)将30只C57BL/6J雄性小鼠随机分为3组:假手术组、模型组、暖心康组(1.65 g·kg^(-1)),每组10只;采用左前降支冠状动脉结扎术复制心肌梗死小鼠模型;灌胃给药,每日1次,连续4周。采用Masson染色法检测心肌组织胶原沉积情况;超声检测小鼠心功能:左室射血分数(LVEF)、收缩末期左室前壁厚度(LVAWS)及舒张末期左室前壁厚度(LVAWD);流式细胞术检测小鼠心脏巨噬细胞分布情况;qPCR法检测心脏组织乳酸脱氢酶A(LDHA)、肉碱棕榈酰转移酶1(CPT-1)、葡萄糖转运蛋白4(GLUT4)、异柠檬酸脱氢酶(IDH)、琥珀酸脱氢酶(SDHa)mRNA表达。(2)按照1.15 g·kg^(-1)剂量给予大鼠暖心康混悬液灌胃,每日2次,持续5 d,制备暖心康含药血清。采用脂多糖(LPS)诱导RAW 264.7细胞构建促炎型巨噬细胞模型。细胞分组:空白血清对照组(含5%空白血清+5%胎牛血清的培养基)、暖心康含药血清组(含5%暖心康含药血清+5%胎牛血清的培养基)、脂多糖组(含5%空白血清+5%胎牛血清的培养基+200μg·mL^(-1)脂多糖)、暖心康含药血清+脂多糖组(含5%暖心康含药血清+5%胎牛血清的培养基+200μg·mL^(-1)脂多糖),干预16 h。采用糖酵解压力测试实验检测RAW 264.7细胞糖酵解水平;qPCR法检测RAW 264.7细胞线粒体丙酮酸转运载体(MPC1)mRNA表达;MitoSox Red荧光染色法检测RAW 264.7细胞线粒体氧化应激损伤程度。结果(1)与假手术组比较,模型组小鼠的心脏胶原纤维蓝染面积明显增加,并伴有室壁变薄,左心室腔增大;LVEF、LVAWS、LVAWD等心功能指标水平均显著降低(P<0.01,P<0.001);小鼠心脏组织中LDHA、CPT-1 mRNA表达明显上调(P<0.05),GLUT4、IDH、SDHa mRNA表达显著下调(P<0.05,P<0.01),CD86染色阳性细胞数量显著增加(P<0.001)。与模型组比较,暖心康组小鼠的心脏�Objective To explore the mechanism of Nuanxinkang Powder(aka.NXK,composed of Ginseng Radix et Rhizoma Rubra and Ilex Pubescens Radix)on improving ventricular remodeling in post-infarction mice based on the"metabolic-inflammatory"network regulating macrophage polarization.Methods①Thirty C57BL/6J male mice were randomly divided into three groups:sham-operation group,model group,and NXK group(1.65 g·kg^(-1)),with 10 mice in each group;the mouse model of myocardial infarction was replicated using left anterior descending coronary artery ligation;and the drug was administered by gavage once a day for 4 consecutive weeks.Masson staining was used to detect collagen deposition in myocardial tissue;ultrasound was used to detect cardiac function in mice:left ventricular ejection fraction(LVEF),left ventricular anterior wall thickness at end-systole(LVAWS)and left ventricular anterior wall thickness at end-diastole(LVAWD);flow cytometry was used to detect distribution of cardiac macrophages in mice;qPCR was used to detect mRNA expressions of lactate dehydrogenase A(LDHA),carnitine palmitoyltransferase 1(CPT-1),glucose transport protein 4(GLUT4),isocitrate dehydrogenase(IDH),and succinate dehydrogenase(SDHa)in heart tissue.②NXK was given 1.15 g·kg^(-1)NXK suspension to rats by gavage twice a day for 5 consecutive days to prepare NXK-containing serum.Lipopolysaccharide(LPS)-induced RAW 264.7 cells were used to construct a pro-inflammatory macrophage model.The cells were grouped into the following groups:blank serum control group(medium containing 5%blank serum+5%fetal bovine serum),NXK drug-containing serum group(medium containing 5%NXK drug-containing serum+5%fetal bovine serum),lipopolysaccharide group(medium containing 5%blank serum+5%fetal bovine serum+200μg·mL^(-1)lipopolysaccharide),NXK drug-containing serum+lipopolysaccharide group(medium containing 5%NXK drugcontaining serum+5%fetal bovine serum+200μg·mL^(-1)lipopolysaccharide),all the groups were intervened for 16 hours.Glycolysis stress test was used to de

关 键 词：暖心康 心肌梗死 心室重构 心肌纤维化 巨噬细胞极化 能量代谢 炎症反应 氧化应激 小鼠 RAW 264.7细胞 

分 类 号：R285.5[医药卫生—中药学]