摘要
研究参附注射液对慢性心力衰竭小鼠的治疗作用及其对巨噬细胞极化的影响。将C57BL/6J小鼠随机分为正常组、造模组。通过腹腔注射异丙肾上腺素(ISO,7.5 mg·kg^(-1),28 d)建立心力衰竭模型,通过检测心功能、N-端前脑钠肽(NT-pro-BNP)判断模型是否复制成功。将造模成功的小鼠随机分为模型组、参附注射液(SFI)组、TAK-242组,并予以相应药物腹腔注射15 d。利用超声心动图评价心脏功能;HE染色检测病理形态;ELISA检测血清NT-proBNP、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)、精氨酸1(Arg-1)水平;流式细胞术检测心脏巨噬细胞相对含量及M1/M2极化状态;qPCR、Western blot检测Toll样受体4(TLR4)/核因子-κB(NF-κB)通路相关mRNA及蛋白表达变化。结果显示,与正常组比较,模型组小鼠左室射血分数(LVEF)、短轴缩短分数(LVFS)降低,左室舒张末内径(LVIDd)、左室收缩末内径(LVIDs)、NT-proBNP、TNF-α、IL-6升高(P<0.01);心脏组织中巨噬细胞数量增多(P<0.05),M1-F4/80^(+)CD86^(+)值上升(P<0.01),M2-F4/80^(+)CD163^(+)值下调(P<0.05);心肌组织TLR4、髓样分化因子88(MyD88)、IκB激酶α(IKKα)、NF-κB p65 mRNA及蛋白表达升高(P<0.05,P<0.01)。与模型组比较,SFI组、TAK-242组小鼠LVEF、LVFS、IL-10、Arg-1升高,LVIDd、LVIDs、NT-proBNP、TNF-α、IL-6降低(P<0.05,P<0.01);心脏F4/80^(+)CD11b^(+)(巨噬细胞)、M1-F4/80^(+)CD86^(+)值下调,M2-F4/80^(+)CD163^(+)值上升(P<0.01,P<0.05);心肌组织TLR4、MyD88、IKKα、NF-κB p65 mRNA及蛋白表达降低(P<0.05,P<0.01)。综上,慢性心力衰竭小鼠存在M1/M2型巨噬细胞极化失衡,以M1型巨噬细胞为主;参附注射液可通过调控TLR4/NF-κB信号通路,促进巨噬细胞向M2极化,抑制M1型巨噬细胞活化,减轻心力衰竭炎症反应。
This study aimed to investigate the therapeutic effect of Shenfu Injection on mice with chronic heart failure(CHF)and its effect on macrophage polarization.C57BL/6J mice were randomly assigned to the normal and model groups.The CHF model was established by intraperitoneal injection of isoproterenol(ISO,7.5 mg·kg^(-1),28 d).The successful modeling was determined by asses-sing the cardiac function and N-terminal pro-brain natriuretic peptide(NT-proBNP).The modeled mice were randomly divided into the model group,Shenfu Injection group,and TAK-242 group,and were injected intraperitoneally with the corresponding drugs for 15 days.Cardiac function was evaluated using echocardiography.Hematoxylin-eosin(HE)staining was used to detect the pathomorphology.Enzyme-linked immunosorbent assay(ELISA)was used to detect the values of serum NT-proBNP,interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),IL-10,and arginase 1(Arg-1).Flow cytometry was applied to detect the relative content and M1/M2 polarization of cardiac macrophages.Quantitative polymerase chain reaction(qPCR)and Western blot were used to detect the changes in the Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)pathway-related mRNA and protein expressions.Compared with the normal group,mice in the model group had lower values of left ventricular ejection fraction(LVEF)and left ventricular fractional shorte-ning(LVFS),higher values of left ventricular internal diastolic end-diastolic(LVIDd),left ventricular internal diastolic end-systolic(LVIDs),NT-proBNP,TNF-α,and IL-6(P<0.01);the number of macrophages increased in cardiac tissues(P<0.05),and the values of M1-F4/80^(+)CD86^(+)were increased(P<0.01),while the values of M2-F4/80^(+)CD163^(+)decreased(P<0.05);the mRNA and protein expressions of TLR4,myeloid differentiation factor 88(MyD88),IκB kinaseα(IKKα),and NF-κB p65 in myocardial tissues were significantly elevated(P<0.05,P<0.01).Compared with the model group,mice in the Shenfu Injection and TAK-242 groups showed elevated LVEF,LVFS,IL-10,and Arg-1 levels,and decreased LVIDd,LVIDs,NT-proBNP,TNF-α,and IL-6 levels(P<0.05,P<0.01);the cardiac F4/80^(+)CD11b^(+)(macrophage)and M1-F4/80^(+)CD86^(+)values were significantly down-regulated,while M2-F4/80^(+)CD163^(+)values were increased(P<0.05,P<0.01);and the mRNA and protein expressions of TLR4,MyD88,IKKα,and NF-κB p65 in myocardial tissues were notably decreased(P<0.05,P<0.01).CHF mice have an imbalance of M1/M2 macrophage polarization,with M1-type macrophages predominating.Shenfu Injection promotes macrophage polarization towards M2,inhibits M1-type macrophage activation,and attenuates inflammatory responses in heart failure by regulating the TLR4/NF-κB signaling pathway.
作者
杨梦
谈宇权
张君宇
张垚
苏丽清
胡志希
胡思远
YANG Meng;TAN Yu-quan;ZHANG Jun-yu;ZHANG Yao;SU Li-qing;HU Zhi-xi;HU Si-yuan(School of Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第13期3574-3582,共9页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(82274412)
湖南省教育厅科学研究项目青年项目(21B0379)
湖南中医药大学校级课题(Z2023XYJB13)。
