| 李海荣,田小溪,校建波,王 琦,黄 潭,张明明,华 瑞.丹参多酚酸盐通过调控肿瘤坏死因子促进大鼠心肌修复的机制研究[J].,2022,(13):2416-2421 |
| 丹参多酚酸盐通过调控肿瘤坏死因子促进大鼠心肌修复的机制研究 |
| Study on the Mechanism of Salvia Miltiorrhiza Polyphenols Promoting Myocardial Repair by Regulating Tumor Necrosis Factor |
| 投稿时间：2021-11-28 修订日期：2021-12-24 |
| DOI：10.13241/j.cnki.pmb.2022.13.003 |
| 中文关键词: 丹参多酚酸盐 TNF-α 血管生成蛋白 细胞迁移 |
| 英文关键词: Salvianolate TNF-α Angiogenic protein Cell migration |
| 基金项目:国家自然科学基金青年科学基金项目（81900338） |
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| 中文摘要: |
| 摘要 目的：探讨丹参多酚酸盐（Sal B）对大鼠损伤后心肌修复的机制。方法：构建新生大鼠心肌细胞H9c2体外缺氧/复氧（H/R）模型，并分组为空白对照组、缺氧/复氧组（模型组）、缺氧/复氧+TNF-α表达质粒组(TNF-α组)和缺氧/复氧+Sal B处理组(Sal B组)。为检测细胞迁移实验，分组为对照组、模型组、H/R+DMSO+Vector组、H/R+Sal B+Vector组、H/R+DMSO+TNF-α组和H/R+Sal B+TNF-α组。通过MTT实验检测各组H9c2细胞活力；免疫荧光检测H9c2心肌细胞中TNF-α细胞表面受体TNFR-1和TNFR-2的表达水平；Western-blot和RT-qPCR检测TNF-α的mRNA和蛋白表达水平以及血管生成蛋白表达水平的影响；Transwell实验检测Sal B对缺氧/复氧处理的H9c2细胞迁移的影响。结果：与对照组相比，模型组H9c2心肌细胞的活力显著下降（P<0.05）；与模型组相比，Sal B组中H9c2心肌细胞的活力显著升高（P<0.05）。免疫荧光检测结果显示，H9c2心肌细胞质膜上TNFR-1和TNFR2均有表达。Western-blot和RT-qPCR结果显示，与模型组相比，Sal B组中H9c2心肌细胞的TNF-α的mRNA和蛋白表达水平均显著升高(P<0.01)，TNF-α组H9c2心肌细胞的TNF-α、Ang-2和VEGF-1蛋白的表达水平均显著升高（P<0.01），Ang-1蛋白表达水平显著降低（P<0.01）。细胞迁移结果显示，与对照组相比，模型组和H/R+DMSO+Vector组H9c2细胞迁移能力显著下降（P<0.01）；与H/R组和H/R+DMSO+Vector组相比，H/R+Sal B+Vector组、H/R+DMSO+TNF-α组和H/R+Sal B+TNF-α组H9c2细胞迁移能力显著升高（P<0.05）。结论：Sal B能够通过上调TNF-α调控血管生成蛋白表达和促进H/R H9c2心肌细胞迁移，从而促进血管生成。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the mechanism of Salvia miltiorrhiza polyphenolate (Sal B) on myocardial repair after injury in rats. Methods: To construct the H9c2 hypoxia/reoxygenation (H/R) model of neonatal rat cardiomyocytes in vitro, and dividing them into blank control group (control group), hypoxia/reoxygenation group (model group), hypoxia/reoxygenation + TNF -α expression plasmid group(TNF-α group) and hypoxia/reoxygenation+Sal B treatment group(Sal B group). To detect cell migration experiments, the cells were divided into control group, model group, H/R+DMSO+Vector group, H/R+Sal B+Vector group, H/R+DMSO+TNF-α group and H/R+ Sal B +TNF- Alpha group. The viability of H9c2 cells in each group were detected by MTT experiment. The expression levels of TNF-α cell surface receptors TNFR-1 and TNFR-2 in H9c2 cardiomyocytes were detected by immunofluorescence. The mRNA and protein expression level of TNF-α and angiogenic protein were detected by Western-blot and RT-qPCR. The effect of Sal B on the migration of H/R H9c2 cells were detected by Transwell experiment. Results: Compared with the control group, the vitality of H9c2 cardiomyocytes in the model group was significantly decreased(P<0.05). Compared with the model group, the vitality of H9c2 cardiomyocytes in the Sal B group was significantly increased(P<0.05). The results of immunofluorescence showed that both TNFR-1 and TNFR2 were expressed on the plasma membrane of H9c2 cardiomyocytes. The results of Western-blot and RT-qPCR showed that compared with model group, the mRNA and protein expression level of TNF-α in H9c2 cell of Sal B group were significantly increased (P<0.01). Compared with the model group, the protein expression levels of TNF-α, Ang-2 and VEGF-1 in H9c2 cell of TNF-α group were significantly increased(P<0.01), the protein expression level of Ang-1 was significantly reduced(P<0.01). The cell migration results showed that compared with the control group, the model group and H/R+DMSO+Vector group H9c2 cell migration ability was significantly weakened(P<0.01). Compared with the model group and H/R+DMSO+Vector group, the migration ability of H9c2 cells in H/R+Sal B+Vector group, H/R+DMSO+TNF-α group and H/R+Sal B+TNF-α group were significantly increased (P<0.05). Conclusion: Sal B promotes angiogenesis by upregulating TNF-α to regulate angiogenic protein expression and promote H/R H9c2 cardiomyocyte migration. |
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