目的 探讨左西孟旦对心肌梗死（myocardial infarction，MI）大鼠心肌纤维化的影响及其作用机制。 方法 通过结扎大鼠左冠状动脉前降支建立MI模型。30只存活的MI大鼠随机分为模型组和左西孟旦组（Levo组），每组15只。假手术组（n=10）的大鼠接受相同的外科手术，只是不进行动脉结扎。Levo组胃灌注左西孟旦4.67 μg·kg-1·d-1治疗28 d。假手术组和模型组大鼠同时给等量蒸馏水。采用二维超声心动图评价心功能，Masson-trichrome评价心肌纤维化程度，RT-qPCR检测COL1A1、COL3A1基因表达，Western blot检测SIRT1/TGF-β1/Smad3信号通路蛋白表达。 结果 超声心动图测量显示，与模型组相比，Levo组大鼠的左室收缩末期内径降低（P<0.05），而射血分数和缩短分数显著升高（P<0.01）。Masson-trichrome显示，左西孟旦治疗可显著减少坏死心肌组织、抑制炎性细胞的浸润并减少胶原蛋白沉积。RT-qPCR分析显示，左西孟旦治疗显著降低了MI大鼠心肌组织中Ⅰ型和Ⅲ型胶原的含量（P<0.01）。Western blot检测显示，左西孟旦治疗可显著上调MI大鼠心肌组织中SIRT1蛋白表达（P<0.01），下调TGF-β1和p-Smad3蛋白表达（P<0.01）。 结论 左西孟旦具有抗纤维化和心保护作用。这些作用可能通过SIRT1/TGF-β1/Smad3信号通路介导。
Abstract
Objective To investigate the effect of levosimendan on myocardial infarction (MI) myocardial fibrosis and its mechanism. Methods MI model was induced by ligation of left anterior descending coronary artery. Thirty surviving MI rats were randomly divided into a model group and a Levo group (15 rats in each group). Rats in the sham group (n=10) underwent the same surgical procedure, except for no arterial ligation. Levosimendan (4.67 μg·kg-1·d-1) was infused into stomach for 28 days in Levo group. At the same time, rats in sham group and model group were given equal amount of distilled water. 2D echocardiography was adopted to evaluate the cardiac function. Masson-trichrome were used to evaluate the degree of myocardial fibrosis. The expression of COL1A1, COL3A1 mRNA and SIRT1/TGF-β1/Smad3 signaling pathway proteins were detected by quantitative reverse transcription PCR (RT-qPCR) and Western blot, respectively. Results Compared with the model group, 2D echocardiography result showed that the LVED of Levo group decreased significantly (P<0.05), while the EF and FS increased significantly (P<0.01). Masson-trichrome result showed that levosimendan treatment significantly reduced necrosis and the content of collagen deposition and inhibited infiltration of inflammatory cells. RT-qPCR analysis result showed that levosimendan treatment significantly reduced the content of type Ⅰ and type Ⅲ collagen in the heart tissue of MI rats (P<0.01). Western blot showed that levosimendan treatment could significantly up-regulate SIRT1 protein expression and down-regulate TGF-β1(60%) and p-Smad3 (47%) protein expression (P<0.01). Conclusions Levosimendan has anti-fibrotic and cardio-protective effects. These effects are potentially mediated by SIRT1/TGF-β1/Smad3 signaling pathway.
关键词
左西孟旦； /
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心肌梗死； /
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大鼠； /
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心肌纤维化； /
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心功能； /
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SIRT
Key words
Levosimendan; /
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Myocardial infarction; /
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Rat; /
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Myocardial fibrosis; /
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Cardiac function /
SIRT
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参考文献
[1] 高鹏荣, 张建, 刘俊杰, 等. 芪参颗粒对心肌梗死后心力衰竭大鼠心肌纤维化的影响[J]. 中医杂志, 2019, 60(13): 1152-1157. DOI: 10.13288/j.11-2166/r.2019.13.015.
[2] 朱灿阳, 吴丹娜, 黎燕峰. 瑞舒伐他汀通过抑制炎症反应改善心肌梗死大鼠心功能及心肌纤维化[J]. 中国临床药理学杂志, 2019, 35(16): 1742-1745. DOI: 10.13699/j.cnki.1001-6821.2019.16.006.
[3] Fan ZB, Fu MH, Xu ZB, et al. Sustained release of a peptide-based matrix metalloproteinase-2 inhibitor to attenuate adverse cardiac remodeling and improve cardiac function following myocardial infarction[J]. Biomacromolecules, 2017, 18(9): 2820-2829. DOI: 10.1021/acs.biomac.7b00760.
[4] 张亮, 李海林. 左西孟旦对顽固性心力衰竭患者心功能的影响[J]. 中国中西医结合急救杂志, 2019, 26(3): 306-308. DOI: 10.3969/j.issn.1008-9691.2019.03.013.
[5] 杨宗达, 简劲峰, 叶华安, 等. 左西孟旦在心功能低下患者冠状动脉旁路移植术后的作用研究[J]. 临床心血管病杂志, 2019, 35(9): 801-804. DOI: 10.13201/j.issn.1001-1439.2019.09.007.
[6] 李秋霞, 慕春言. 伊伐布雷定联合左西孟旦治疗慢性心力衰竭的临床研究[J]. 现代药物与临床, 2019, 34(6): 1630-1634. DOI: 10.7501/j.issn.1674-5515.2019.06.006.
[7] Aminzadeh A, Mehrzadi S. Cardioprotective effect of levosimendan against homocysteine-induced mitochondrial stress and apoptotic cell death in H9C2[J]. Biochem Biophys Res Commun, 2018, 507(1-4): 395-399. DOI: 10.1016/j.bbrc.2018.11.049.
[8] Hadi K, Onur K, Vikram P, et al. Fibroblast-specific TGF-β-Smad2/3 signaling underlies cardiac fibrosis[J]. J Clin Invest, 2017, 127(10): 3770-3783. DOI: 10.1172/JCI94753.
[9] Islam SS, Mokhtari RB, El Hout Y, et al. TGF-β1 induces EMT reprogramming of porcine bladder urothelial cells into collagen producing fibroblasts-like cells in a Smad2/Smad3-dependent manner[J]. J Cell Commun Signal, 2014, 8(1): 39-58. DOI: 10.1007/s12079-013-0216-4.
[10] Kong P, Christia P, Frangogiannis NG. The pathogenesis of cardiac fibrosis[J]. Cell Mol Life Sci, 2014, 71(4): 549-574. DOI: 10.1007/s00018-013-1349-6.
[11] 翟铁, 郝凤杰, 田雪品, 等. 过表达SIRT1基因通过PI3K/AKT信号通路抑制高糖刺激心肌H9c2细胞凋亡和活性氧水平[J]. 中国病理生理杂志, 2019, 35(5): 129-133. DOI: 10.3969/j.issn.1000-4718. 2019. 05. 018.
[12] Li ST, Zhu ZX, Xue M, et al. Fibroblast growth factor 21 protects the heart from angiotensin II-induced cardiac hypertrophy and dysfunction via SIRT1[J]. Biochim Biophys Acta Mol Basis Dis, 2019, 1865(6): 1241-1252. DOI: 10.1016/j.bbadis.2019.01.019.
[13] Price NL, Gomes AP, Ling AJ, et al. SIRT1 is required for AMPK activation and the beneficial effects of resveratrol on mitochondrial function[J]. Cell Metab, 2012, 15(5): 675-690. DOI: 10.1016/j.cmet.2012.04.003.
[14] 郝洲华, 卢晓明, 孟浦, 等. miR-129-3p靶向Smad3在TGF-β诱导NIH3T3细胞转分化为肌成纤维细胞过程中抑制细胞活力与迁移[J]. 中国病理生理杂志, 2019, 35(9): 1668-1675. DOI: 10.3969/j.issn.1000-4718.2019.09.021.
[15] Lin YC, Luo HY, Wang X, et al. Flavanones from sedum sarmentosum bunge alleviate CCl4-induced liver fibrosis in rats by targeting TGF-β1/TβR/Smad Ppathway in turn inhibiting epithelial mesenchymal transition[J]. Evid Based Complement Alternat Med, 2018, 3(8): 3080837. DOI: 10.1155/2018/3080837.
基金
2020年度河北省医学科学课题研究计划（20200498）
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