目的 考察黄芩苷通过激活CPT1、维持线粒体动力学稳定缓解心肌细胞损伤的作用机制。方法 通过使用氧气和葡萄糖剥夺（OGD）构建AC16、H9c2、HL-1细胞损伤模型。用黄芩苷（0.1、1.0、10.0 μg/mL）处理模型细胞，考察细胞增殖及凋亡情况。通过RNA测序技术检测黄芩苷治疗前后，细胞转录谱的变化，并通过RT-qPCR、Western blotting对线粒体动力学相关标志物PPARGC1A、Drp1、OPA1、MFN1、MFN2、ESRRA、NQO1以及p-DRP1、DRP1、MFN1、NRF1、TFAM进行验证，并使用透射电镜对线粒体形貌进行考察。最后探究黄芩苷对其靶点CPT1的作用。结果 与模型组比较，黄芩苷（1.0、10.0μg/mL）组处理后的细胞活性有显著升高，细胞凋亡率显著降低（P＜0.01、0.001）。RNA测序及Western blotting结果表明，黄芩苷可显著提高OPA1、MFN1、ESRRA、NQO1 mRNA水平以及MFN1、NRF1的蛋白表达水平（P＜0.001）。黄芩苷处理后，AC16细胞内的空泡体积明显减小，线粒体数量和线粒体嵴均有所恢复。黄芩苷能在H9c2和HL-1细胞原位和体外直接激活CPT1，提高其活性（P＜0.001）。结论 黄芩苷均能缓解心肌细胞活性的下降和凋亡的增加，并通过激活CPT1、维持线粒体动力学稳定缓解心肌细胞损伤。

Objective To investigate the mechanism of baicalin alleviates cardiomyocyte injury by activating CPT1 and maintaining the stability of mitochondrial dynamics. Methods The injury models of AC16, H9c2, and HL-1 cells were constructed by using oxygen and glucose deprivation (OGD). Model cells were treated with baicalin (0.1, 1.0, and 10.0 μg/mL) to investigate cell proliferation and apoptosis. The changes of cell transcriptome profiles before and after baicalin treatment were detected by RNA sequencing technology. The mitochondrial dynamics-related markers PPARGC1A, Drp1, OPA1, MFN1, MFN2, ESRRA, NQO1, as well as p-DRP1, DRP1, MFN1, NRF1, and TFAM were verified by RT-qPCR and Western blotting. The morphology of mitochondria was investigated by transmission electron microscopy. Finally, the effect of baicalin on its target CPT1 were explored. Results Compared with the model group, the cell viability in the baicalin (1.0, 10.0 μg/mL) groups was significantly increased, and the apoptosis rate was significantly decreased (P < 0.01, 0.001). The results of RNA sequencing and Western blotting indicated that baicalin could significantly increase the mRNA levels of OPA1, MFN1, ESRRA, and NQO1, as well as the protein expression levels of MFN1 and NRF1 (P < 0.001). After treatment with baicalin, the vacuole volume in AC16 cells was significantly reduced, and both the number of mitochondria and mitochondrial cristae were restored. Baicalin can directly activate CPT1 in H9c2 and HL-1 cells in situ and in vitro, and enhance their activity (P < 0.001). Conclusion Baicalin can alleviate the decline in myocardial cell activity and the increase in apoptosis, and relieve myocardial cell injury by activating CPT1 and maintaining the stability of mitochondrial dynamics.

R966

国家自然科学基金青年科学基金项目（82003885）