目的 探究松果菊苷对阿霉素（doxorubicin，DOX）诱导的心肌细胞损伤的影响及作用机制。方法 不同质量浓度（5、10、20 μg/mL）的松果菊苷作用于DOX处理的大鼠H9c2心肌细胞48 h，采用CCK-8法测定细胞活性，通过流式细胞术分析细胞凋亡情况；采用Western blotting检测剪切型半胱氨酸天冬氨酸蛋白酶-3（cleaved cystein-asparate protease-3，cleaved Caspase-3）和B细胞淋巴瘤-2相关的X蛋白（B-cell lymphoma-2 associated X protein，Bax）表达；检测细胞内丙二醛（malondialdehyde，MDA）水平及超氧化物歧化酶（superoxide dismutase，SOD）、过氧化氢酶（catalase，CAT）活性和细胞培养上清中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin-6，IL-6）水平；采用qRT-PCR检测miR-221-3p表达。分别转染miR-221-3p抑制剂（anti-miR-221-3p）或模拟物（miR-221-3p mimics）至H9c2细胞，用DOX处理48 h后，观察miR-221-3p对DOX诱导的H9c2细胞活性、凋亡、氧化应激和炎症反应的影响。结果 松果菊苷可提高DOX处理的H9c2细胞活性和SOD活性（P＜0.05），抑制DOX诱导的H9c2细胞凋亡及cleaved Caspase-3、Bax蛋白表达（P＜0.05），降低MDA、TNF-α和IL-6水平（P＜0.05）。下调miR-221-3p后，DOX处理的H9c2细胞活性和SOD活性升高（P＜0.05），细胞凋亡率、cleaved Caspase-3和Bax蛋白表达、MDA、TNF-α和IL-6水平均降低（P＜0.05）。松果菊苷降低了DOX处理的H9c2细胞中miR-221-3p表达（P＜0.05），上调miR-221-3p表达逆转了松果菊苷对DOX处理的H9c2细胞凋亡、氧化应激和炎性因子水平的作用（P＜0.05）。结论 松果菊苷可显著提高经DOX干预的H9c2细胞增殖能力，同时有效抑制细胞凋亡进程、减轻氧化应激损伤及炎症因子释放。其作用机制可能与调控miR-221-3p的表达有关。

Objective To explore the effect and mechanism of echinacoside on doxorubicin (DOX)-induced cardiomyocyte damage. Methods Different concentrations (5, 10, 20 μg/mL) of echinacoside were used to act on DOX-treated rat H9c2 cardiomyocytes for 48 h. The cell viability was determined using CCK-8 assay, cell apoptosis was analyzed by flow cytometry; Western blotting was used to detect the expressions of cleaved cysteine aspartate protease-3 (cleaved Caspase-3) and B-cell lymphoma-2 associated X protein (Bax); Level of intracellular malondialdehyde (MDA) and activities of superoxide dismutase (SOD), catalase (CAT), as well as levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) in cell culture supernatant were measured; qRT-PCR was used to detect the expression of miR-221-3p. miR-221-3p inhibitors (anti-miR-221-3p) or mimics (miR-221-3p mimics) were transfected into H9c2 cells, and after 48 h of DOX treatment, the effects of miR-221-3p on viability, apoptosis, oxidative stress and inflammatory response in DOX-induced H9c2 cells were observed. Results Echinacoside could increase the viability and SOD activity of H9c2 cells treated with DOX (P < 0.05), inhibit apoptosis of DOX-induced H9c2 cells and expressions of cleaved Caspase-3 and Bax proteins (P < 0.05), and reduce the levels of MDA, TNF-α and IL-6 (P < 0.05). After down-regulating miR-221-3p, the viability and SOD activity of H9c2 cells treated with DOX were increased (P < 0.05), while the apoptosis rate, expressions of cleaved Caspase-3 and Bax proteins, levels of MDA, TNF-α and IL-6 were decreased (P < 0.05). Echinacoside reduced the expression of miR-221-3p in H9c2 cells treated with DOX (P < 0.05), while up-regulating miR-221-3p expression reversed the effects of echinacoside on apoptosis, oxidative stress and inflammatory factor levels in H9c2 cells treated with DOX (P < 0.05). Conclusion Echinacoside could significantly enhance the proliferation ability of H9c2 cells intervened by DOX, while effectively inhibiting the process of cell apoptosis, reducing oxidative stress damage and inflammatory cytokine release. Its mechanism may be related to the regulation of miR-221-3p expression.

R285.5

河南省卫生健康委员会科研项目（LHGJ20210036）