浙江省中医药研究院实验动物中心,杭州 310007
Department of Experimental Animals, Zhejiang Academy of Traditional Chinese Medicine, Hangzhou 310007, China
目的 研究土茯苓总黄酮(Smilax glabra flavonoids,SGF)对异丙肾上腺素( isoproterenol,ISO)诱导的小鼠心肌肥大和心脏炎症的作用及机制。 方法 C57 / 6J 小鼠随机分为正常组、ISO 模型组(1. 25 mg / (kg·d))、SGF低剂量组(50 mg / (kg·d))、SGF 高剂量组(100 mg / (kg·d))。 SGF 低剂量组和 SGF 高剂量组均在造模前预防给药7 d,之后各组进行 ISO 皮下注射,连续注射 7 d 后进行超声心动图测定;眼眶取血分离血清,检测血清中 N 端脑钠肽前体(N-terminal brain natriuretic peptide precursor,NT-proBNP)及炎症因子白细胞介素-1β( interleukin-1 beta,IL1β)、肿瘤坏死因子 α(tumor necrosis factor-α,TNF-α)、白介素 6( interleukin-6,IL-6)含量;收集心肌标本,观察心肌组织的病理学改变,二氢乙锭(dihydroethidium,DHE)染色法检测心肌活性氧( reactive oxygen species,ROS)水平,检测心脏相关肥大基因 mRNA 水平变化以及 NOD 样受体热蛋白结构域蛋白 3 / 半胱天冬酶-1/ 白介素 18(NOD-like receptor thermal protein domain associated protein 3 / cysteinyl aspartate specific proteinase-1/ interleukin-18, NLRP3 /Caspase-1/ IL-18)炎症级联通路上关键因子蛋白水平表达的变化。 结果 SGF 预防给药可降低超声心动图中的左室舒张末期室间隔厚度(left ventricular end-diastolic inter ventricular septal thickness,IVSd)、左室收缩末期室间隔厚度(left ventricular end-systolic interventricular septal thickness,IVSs)和射血分数(ejection fraction,EF),提高左室舒张末期内径( left ventricular end-diastolic diameter,LVIDd)、左室收缩末期内径( left ventricular end-systolic diameter,LVIDs)和左心室收缩末期容积( left ventricular end systolic volume,LVESV),降低血中 IL-1β、IL-6、TNF-α 和 NTproBNP 含量,抑制心脏相关肥大基因心房钠尿肽( atrial natriuretic peptide,ANP)、脑钠肽( brain natriuretic peptide,BNP)、β-肌球蛋白重链( beta-myosin heavy chain,β-MHC) mRNA 表达水平上调,抑制心肌 ROS 水平增加,下调NLRP3 / Caspase-1/ IL-18 炎症级联通路上关键因子 NLRP3、Caspase-1(p20)和 IL-18 的蛋白表达,改善心肌细胞肥大和排列紊乱的情况,减少心肌纤维外的成纤维细胞增多,减轻炎症细胞浸润和心肌组织纤维化情况。 结论 SGF可改善 ISO 诱导的小鼠心肌肥大和心脏炎症,可能通过 NLRP3 / Caspase-1/ IL-18 炎症级联通路起作用。
Objective To study the effects and mechanisms of Smilax glabra flavonoids ( SGF) on myocardial hypertrophy and cardiac inflammation induced by isoproterenol ( ISO) in mice. Methods C57 / 6J mice were randomly divided into a normal group, ISO model group (1. 25 mg / ( kg·d)), SGF low-dose group (50 mg / ( kg·d)), and SGF high-dose group (100 mg / ( kg·d)). The SGF groups were given prophylactic SGF for 7 days before modeling, then subcutaneous ISO injection was given to each group, and echocardiography was performed after continuous injection for 7days. Serum was separated from orbital blood, and the NT-proBNP and inflammatory factor ( IL-1β, IL-6, and TNF-α) contents of the blood were detected. Myocardial specimens were collected, and pathological changes to myocardial tissue were observed. Myocardial ROS levels were detected by DHE staining. The mRNA levels of heart-related hypertrophy genes and changes in the expression of key proteins in the inflammatory pathway of NLRP3 / Caspase-1/ IL-18 in myocardial tissue were detected. Results SGF prophylactic administration decreased IVSd, IVSs, and EF in echocardiography and increased LVIDs, LVIDd, and LVESV. The IL-1β, IL-6, TNF-α, and NT-proBNP contents of blood decreased, and the mRNA expression levels of the heart-related hypertrophy genes for ANP, BNP, and β-MHC were subdued. The increase in myocardial ROS levels was also inhibited. The protein expression of NLRP3, Caspase-1 (p20), and IL-18, which are key factors in the NLRP3 / caspase-1/ IL-18 inflammatory pathway, was down-regulated in myocardial tissue. The hypertrophy and disordered arrangement of cardiomyocytes were improved, the increase in fibroblast numbers outside myocardial fibers was reduced, and the infiltration of inflammatory cells and fibrosis of myocardial tissue were alleviated. Conclusions SGF can improve ISO-induced myocardial hypertrophy and cardiac inflammation in mice and may act through the NLRP3 /Caspase-1/ IL-18 inflammatory pathway.
富丹婷,施佳君,杨钦钦,郑纯威,黄强,陈宇.土茯苓总黄酮改善 ISO 诱导的小鼠心肌肥大和心脏炎症的预防作用及机制[J].中国实验动物学报,2024,32(8):1023~1031.
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