| 高兰英,贾 彬,文 健,庄 艳,罗 勇.竹节参皂苷IVa减轻异丙肾上腺素诱导的小鼠心肌纤维化作用机制研究[J].,2021,(21):4055-4061 |
| 竹节参皂苷IVa减轻异丙肾上腺素诱导的小鼠心肌纤维化作用机制研究 |
| Effect of Chikusetsusaponin IVa on Myocardial Fibrosis Induced by Isoproterenol in Mice |
| 投稿时间：2021-04-27 修订日期：2021-05-23 |
| DOI：10.13241/j.cnki.pmb.2021.21.011 |
| 中文关键词: 竹节参皂苷IVa 异丙肾上腺素 小鼠 心肌纤维化 AMPK/mTOR/ULK1 |
| 英文关键词: Chikusetsusaponin IVA Isoproterenol Mice Myocardial fibrosis AMPK/mTOR/ULK1 |
| 基金项目:四川省医学科研青年创新课题(Q14013) |
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| 中文摘要: |
| 摘要 目的：探讨竹节参皂苷（IVa）减轻异丙肾上腺素（ISO）诱导的小鼠心肌纤维化作用机制。方法：白变种实验室老鼠（Balb/C）小鼠40只并随机分为4组：正常对照组（n=10）、ISO模型组（n=10）、IVa低剂量组（n=10）、IVa高剂量组（n=10）。采用皮下注射ISO构建小鼠心肌纤维化模型，IVa剂量组在建模同时给予IVa治疗，正常组给予等量生理盐水。采用马松（Masson）三色标准和HE染色方法分析评估心脏组织形态学和胶原沉积。采用小麦胚芽凝集素（WGA）染色法测定心肌细胞面积。蛋白免疫印迹试验检测细胞自噬相关标志物（LC3-Ⅱ、Beclin1和p62）和腺苷单磷酸活化蛋白激酶（AMPK）/哺乳动物雷帕霉素靶蛋白（mTOR）/自噬激活激酶1（ULK1）信号通路相关标志物。采用酶联免疫吸附法（ELISA）检测血清中血管紧张素II（Ang II）和I型前胶原羧基末端肽（PICP）的含量。结果：高剂量IVa（15mg/kg）治疗后，HW/BW和LVW/BW较ISO模型组升高，而血清中Ang II和PICP含量降低。IVa以剂量依赖的方式减轻心肌细胞在心脏组织中的损伤。皮下注射ISO后，心肌间质内胶原沉积明显，IVa治疗后胶原沉积明显减少。IVa可有效降低ISO诱导的小鼠心肌细胞面积大小。IVa能有效抑制ISO诱导的LC3-Ⅱ和Beclin1蛋白降低，减少p62蛋白增多。AMPK直接磷酸化ULK1（Ser555），通过抑制mTOR磷酸化间接抑制ULK1（Ser757）磷酸化，均参与了ISO诱导的心肌纤维化小鼠自噬活性的降低；此外，IVa低剂量组和IVa高剂量组均显著增加了AMPK磷酸化，并抑制了mTOR磷酸化，降低了ULK1(Ser757)磷酸化。结论：IVa通过AMPK/mTOR/ULK1途径激活自噬，减轻了ISO诱导的心肌纤维化。表明IVa是一种潜在的抗心肌纤维化候选药物，是治疗心脏病的潜在药物靶点。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the mechanism of Chikusetsusaponin (IVa) on myocardial fibrosis induced by isoproterenol (ISO) in mice. Methods: 40 BALB/c mice were randomly divided into four groups: normal control group (n=10), ISO model group (n=10), IVa low-dose group (n=10) and IVa high-dose group (n=10). The mouse myocardial fibrosis model was established by subcutaneous injection of ISO. The IVa dose group was treated with IVa at the same time, and the normal group was treated with the same amount of normal saline. Masson trichrome standard and HE staining were used to analyze and evaluate cardiac histomorphology and collagen deposition. The area of cardiomyocytes was measured by wheat germ agglutinin(WGA) staining. Autophagy related markers (LC3-Ⅱ, Beclin1 and p62) and AMP-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)/Unc-like kinase1(ULK1) signal pathway related markers were detected by Western blot. The contents of angiotensin II(ANG II) and type I procollagen carboxy terminal peptide(PICP) in serum were detected by enzyme-linked immunosorbent assay(ELISA). Results: After high-dose IVa (15 mg/kg), HW/BW and LVW/BW increased compared with ISO model group, while the contents of Ang II and PICP in serum decreased. IVa alleviates the injury of cardiomyocytes in heart tissue in a dose-dependent manner. After subcutaneous injection of ISO, collagen deposition in myocardial stroma was obvious, and collagen deposition decreased significantly after IVa treatment. IVa can effectively reduce the area and size of mouse cardiomyocytes induced by ISO. IVa can effectively inhibit the decrease of LC3-Ⅱ and Beclin1 protein and the increase of p62 protein induced by ISO. AMPK directly phosphorylates ULK1 (ser555) and indirectly inhibits ULK1 (ser757) phosphorylation by inhibiting mTOR phosphorylation, which were involved in the reduction of autophagy activity in ISO induced myocardial fibrosis mice. In addition, both low-dose and high-dose IVa groups significantly increased AMPK phosphorylation, inhibited mTOR phosphorylation and decreased ULK1(ser757) phosphorylation. Conclusion: IVa activates autophagy through AMPK/mTOR/ULK1 pathway and reduces ISO induced myocardial fibrosis. It shows that IVa is a potential candidate drug for anti my ocardial fibrosis and a potential drug target for the treatment of heart disease. |
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