目的 考察黃芩苷通過(guò)激活CPT1、維持線粒體動(dòng)力學(xué)穩(wěn)定緩解心肌細(xì)胞損傷的作用機(jī)制。方法 通過(guò)使用氧氣和葡萄糖剝奪（OGD）構(gòu)建AC16、H9c2、HL-1細(xì)胞損傷模型。用黃芩苷（0.1、1.0、10.0 μg/mL）處理模型細(xì)胞，考察細(xì)胞增殖及凋亡情況。通過(guò)RNA測(cè)序技術(shù)檢測(cè)黃芩苷治療前后，細(xì)胞轉(zhuǎn)錄譜的變化，并通過(guò)RT-qPCR、Western blotting對(duì)線粒體動(dòng)力學(xué)相關(guān)標(biāo)志物PPARGC1A、Drp1、OPA1、MFN1、MFN2、ESRRA、NQO1以及p-DRP1、DRP1、MFN1、NRF1、TFAM進(jìn)行驗(yàn)證，并使用透射電鏡對(duì)線粒體形貌進(jìn)行考察。最后探究黃芩苷對(duì)其靶點(diǎn)CPT1的作用。結(jié)果 與模型組比較，黃芩苷（1.0、10.0μg/mL）組處理后的細(xì)胞活性有顯著升高，細(xì)胞凋亡率顯著降低（P＜0.01、0.001）。RNA測(cè)序及Western blotting結(jié)果表明，黃芩苷可顯著提高OPA1、MFN1、ESRRA、NQO1 mRNA水平以及MFN1、NRF1的蛋白表達(dá)水平（P＜0.001）。黃芩苷處理后，AC16細(xì)胞內(nèi)的空泡體積明顯減小，線粒體數(shù)量和線粒體嵴均有所恢復(fù)。黃芩苷能在H9c2和HL-1細(xì)胞原位和體外直接激活CPT1，提高其活性（P＜0.001）。結(jié)論 黃芩苷均能緩解心肌細(xì)胞活性的下降和凋亡的增加，并通過(guò)激活CPT1、維持線粒體動(dòng)力學(xué)穩(wěn)定緩解心肌細(xì)胞損傷。

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

國(guó)家自然科學(xué)基金青年科學(xué)基金項(xiàng)目（82003885）