| 王审,许铭明,叶紫恒,屠俊杰.涤痰活血舒痹汤上调microRNA-148a表达减轻心肌缺血再灌注损伤的分子机制研究[J].浙江中西医结合杂志,2022,32(12): |
| 涤痰活血舒痹汤上调microRNA-148a表达减轻心肌缺血再灌注损伤的分子机制研究 |
| Molecular mechanism of Ditan Huoxue Shubi Decoction up-regulating microRNA-148a expression to alleviate myocardial ischemia-reperfusion injury |
| 投稿时间：2022-02-09 修订日期：2022-09-27 |
| DOI： |
| 中文关键词: 心肌缺血再灌注损伤 涤痰活血舒痹汤 miR-148a TXNIP 自噬 |
| 英文关键词:Myocardial ischemia reperfusion injury Ditan Huoxue Shubi Decoction miR-148a TXNIP Autophagy. |
| 基金项目:浙江省中医药科技计划项目(2022ZB149) |
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| 中文摘要: |
| 目的 旨在探究涤痰活血舒痹汤（Ditan Huoxue Shubi Decoction，DHS）与microRNA-148a（miR-148a）的调控关系以及其在心肌缺血再灌注损伤中的分子作用机制。方法 将H9c2细胞培养后分为对照组、缺氧/复氧（hypoxia/reoxygenation，H/R）组、H/R+空白血清组、H/R+DHS含药血清组、H/R+DHS含药血清+miR-148a拮抗剂组、H/R+DHS含药血清+OE-NC组和H/R+DHS含药血清+TXNIP过表达质粒组。采用CCK-8检测H9c2细胞增殖，TUNEL染色检测H9c2细胞凋亡。运用Western blot检测H9c2细胞中凋亡和自噬相关蛋白表达。利用 qRT-PCR检测miR-148a的表达。Western blot检测靶蛋白TXNIP的表达。结果 与对照组比较，H/R组H9c2细胞活力下降[（97.75±5.58）%比（69.72±8.80）%，P < 0.01]、细胞凋亡和自噬水平上升。与H/R+空白血清组比较，H/R+DHS含药血清组miRNA-148a表达上调[（1.04±0.25）比（2.45±0.39），P < 0.01]，H9c2细胞活力增加[（97.75±7.94）%比（135.98±10.45）%，P < 0.01]并且细胞凋亡和自噬显著下降（P < 0.05）。H/R+DHS含药血清+miR-148a拮抗剂组细胞内miRNA-148a表达下调[（2.18±0.25）比（0.66±0.12），P < 0.001]，同时靶基因TXNIP表达上调。与H/R+DHS含药血清+OE-NC组比较，H/R+DHS含药血清+TXNIP过表达质粒组的TXNIP表达水平、细胞凋亡和自噬水平显著上升（P < 0.05）。结论 涤痰活血舒痹汤能够减轻H/R引起的心肌细胞H9c2细胞的损伤，其作用机制可能是通过升高miR-148a水平而减低TXNIP表达来降低损伤的心肌细胞自噬和凋亡水平来实现的。 |
| 英文摘要: |
| ABSTRACT Objective This article aims to explore the regulatory relationship between Ditan Huoxue Shubi Decoction (DHS) and microRNA-148a (miR-148a) and its molecular mechanism in myocardial ischemia-reperfusion injury. Methods H9c2 cells were cultured and divided into control group, hypoxia/reoxygenation (H/R) group, H/R + blank serum group, H/R+DHS medicated serum group, H/R+DHS medicated serum+miR-148 a antagonist group, H/R+DHS medicated serum+OE-NC group, and H/R+DHS medicated serum+TXNIP overexpression plasmid group. CCK-8 was used to detect the proliferation of H9c2 cells, and TUNEL staining was used to detect the apoptosis of H9c2 cells. Western blot was used to detect the expression of apoptosis and autophagy-related proteins in H9c2 cells. qRT-PCR was used to detect the expression of miR-148a. The expression of target protein TXNIP was detected by Western blot. Results Compared with control group, H9c2 cell viability decreased [(97.75±5.58) % vs (69.72±8.80) %, P < 0.01], apoptosis and autophagy increased in H/R group. Compared with the H/R+blank serum group, the expression of miRNA-148a in the H/R+DHS drug-containing serum group was up-regulated [(1.04±0.25) vs (2.45±0.39) , P < 0.01], the cell viability of H9c2 was increased [(97.75±7.94) % vs (135.98±10.45) %, P < 0.01], and the apoptosis and autophagy were significantly decreased (P < 0.05). The expression of miRNA-148a in the H/R+DHS drug-containing serum+miR-148a antagonist group was down-regulated [(2.18±0.25) vs (0.66±0.12) , P < 0.001], and the expression of target gene TXNIP was up-regulated. Compared with the H/R+DHS drug-containing serum+OE-NC group, the expression of TXNIP in the H/R+DHS drug-containing serum+TXNIP overexpression plasmid group, and the levels of apoptosis and autophagy were significantly increased（P < 0.05）. Conclusion Ditan Huoxue Shubi Decoction can alleviate the damage of cardiomyocytes H9c2 cells caused by H/R, and its mechanism may be achieved by increasing the level of miR-148a and reducing the expression of TXNIP to reduce the autophagy and apoptosis of damaged cardiomyocytes. |
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