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陆哲雯, 刘莉, 蒋啸, 李鑫辉.丹参通络解毒汤调控METTL3介导的m6A修饰对缺氧/复氧CMECs的保护作用[J].湖南中医药大学学报英文版,2025,45(3):438-444.[Click to copy
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| 丹参通络解毒汤调控METTL3介导的m6A修饰对缺氧/复氧CMECs的保护作用 |
| 陆哲雯,刘莉,蒋啸,李鑫辉 |
| (湖南中医药大学中医学院, 湖南 长沙 410208) |
| 摘要: |
| 目的 探讨丹参通络解毒汤对缺氧/复氧（H/R）损伤大鼠心肌微血管内皮细胞（CMECs）的保护作用及甲基转移酶样3（METTL3）的影响。方法 建立H/R损伤CMECs模型，将细胞分为空白组、模型组、中药组、过表达组、中药+过表达组、空载组，ELISA检测各组炎症因子IL-1β和IL-10水平，流式细胞术检测各组细胞凋亡情况，Western blot检测各组METTL3蛋白表达水平，比色法检测各组m6A及肌酸激酶CK水平。结果 与空白组比较，模型组METTL3蛋白表达与m6A水平均升高（P＜0.01），CMECs凋亡率升高（P＜0.01），IL-1β、CK含量升高（P＜0.01），IL-10含量降低（P＜0.01）；与模型组比较，中药组METTL3蛋白表达与m6A水平均降低（P＜0.05，P＜0.01），CMECs凋亡率显著降低（P＜0.01），IL-1β、CK含量显著降低（P＜0.01），IL-10含量显著升高（P＜0.01）；与模型组比较，过表达组METTL3蛋白表达升高（P＜0.01），CMECs凋亡率升高（P＜0.01），IL-1β、CK含量升高（P＜0.01，P＜0.05），IL-10含量降低（P＜0.01）；与过表达组相比，中药+过表达组METTL3蛋白表达降低（P＜0.01），CMECs凋亡率降低（P＜0.01），IL-1β、CK含量降低（P＜0.01，P＜0.05），IL-10含量显著升高（P＜0.01）。结论 丹参通络解毒汤能降低炎症反应，抑制细胞凋亡，对缺氧/复氧CMECs有保护作用，其机制可能与抑制METTL3表达，从而下调m6A水平有关。 |
| 关键词: 丹参通络解毒汤 甲基转移酶样3 m6A修饰 心肌缺血再灌注损伤 心肌微血管内皮细胞 |
| DOI：10.3969/j.issn.1674-070X.2025.03.007 |
| Received:November 30, 2024 |
| 基金项目:国家自然科学基金面上项目（82074392）；湖南省自然科学基金区域联合基金项目（2023JJ50036）；湖南省中医药管理局重点项目（A2023013）；湖南省教育厅科学研究项目（23A0283）。 |
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| Protective effects of Danshen Tongluo Jiedu Decoction on hypoxia/reoxygenation-injured CMECs through regulating METTL3-mediated m6A modification |
| LU Zhewen, LIU Li, JIANG Xiao, LI Xinhui |
| (School of Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To explore the protective effects of Danshen Tongluo Jiedu Decoction (DSTLJDD) on cardiac microvascular endothelial cells (CMECs) of rats with hypoxia/reoxygenation (H/R) injury and its influence on methyltransferase-like 3 (METTL3). Methods The H/R-injured CMECs model was established, and the cells were divided into blank group, model group, Chinese medicine (CM) group, overexpression group, CM+overexpression group, and empty vector group. The levels of inflammatory factors IL-1β and IL-10 in each group were determined by ELISA, cell apoptosis was measured by flow cytometry, the protein expression level of METTL3 was checked by Western blot, and the content of m6A and creatine kinase (CK) were examined by colorimetry. Results Compared with the blank group, the protein expression of METTL3 and the level of m6A in the model group increased (P<0.01), the apoptosis rate of CMECs increased (P<0.01), the content of IL-1β and CK increased (P<0.01), while the content of IL-10 decreased (P<0.01). Compared with the model group, the protein expression of METTL3 and the level of m6A in the CM group decreased (P<0.05, P<0.01), the apoptosis rate of CMECs markedly decreased (P<0.01), the content of IL-1β and CK significantly decreased (P<0.01), while the content of IL-10 notably increased (P<0.01). Compared with the model group, the protein expression of METTL3 in the overexpression group increased (P<0.01), the apoptosis rate of CMECs increased (P<0.01), the content of IL-1β and CK increased (P<0.01, P<0.05), while the content of IL-10 decreased (P<0.01). Compared with the overexpression group, the protein expression of METTL3 in the CM+overexpression group decreased (P<0.01), the apoptosis rate of CMECs decreased (P<0.01), the content of IL-1β and CK decreased (P<0.01, P<0.05), while the content of IL-10 significantly increased (P<0.01). Conclusion DSTLJDD can reduce inflammatory responses, inhibit cell apoptosis, and exert protective effects on H/R-injured CMECs. Its mechanism may be related to the suppression of METTL3 expression, thereby downregulating m6A level. |
| Key words: Danshen Tongluo Jiedu Decoction methyltransferase-like 3 m6A modification myocardial ischemia-reperfusion injury cardiac microvascular endothelial cells |
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