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黄芩素和汉黄芩素抑制肝癌细胞能量代谢的作用机制差异研究
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| 篇名： | 黄芩素和汉黄芩素抑制肝癌细胞能量代谢的作用机制差异研究 |
| TITLE： | Study on mechanism difference of baicalein and wogonin inhibiting energy metabolism of hepatoma cells |
| 摘要： | 目的 探究黄芩素和汉黄芩素抑制肝癌细胞能量代谢的作用机制差异。方法将人肝癌HepG2细胞分为空白对照组（不给药）、黄芩素和汉黄芩素不同剂量组（均为1.25、2.5、5、10、20μmol/L），采用MTT法检测黄芩素和汉黄芩素对细胞活力的影响。将HepG2细胞分为空白对照组（不给药）、黄芩素组、汉黄芩素组，给药培养后，采用增强型ATP试剂盒检测细胞ATP浓度，采用糖酵解和线粒体压力测试试剂盒评价糖酵解及线粒体能量代谢水平；采用分子对接预测黄芩素和汉黄芩素与能量代谢关键酶的亲和力大小，筛选亲和力较高的能量代谢关键酶；采用Westernblot法检测能量代谢关键酶的表达情况。结果在2.5～20μmol/L剂量范围内，黄芩素和汉黄芩素的半数抑制浓度分别为12.84、24.09μmol/L，且黄芩素1.25μmol/L、汉黄芩素2.5μmol/L对细胞活力基本无影响，筛选其为后续实验的给药剂量。与空白对照组比较，黄芩素组、汉黄芩素组HepG2细胞ATP浓度显著降低（P＜0.05）；汉黄芩素对HepG2细胞基础酸化率的抑制作用显著强于黄芩素（P＜0.05），但两者对基础耗氧率的抑制作用均无显著差异（P＞0.05）；黄芩素与丙酮酸激酶M2、线粒体酶复合物Ⅰ（CⅠ）、CⅡ、CⅣ结合力较强，而汉黄芩素只与丙酮酸激酶M2的结合力较强；汉黄芩素能显著下调己糖激酶、磷酸果糖激酶、丙酮酸激酶M2、CⅠ、CⅡ、CⅣ蛋白的表达水平（P＜0.05），而黄芩素对上述酶的调节作用无统计学意义（P＞0.05）。结论黄芩素和汉黄芩素均能抑制肝癌HepG2细胞能量代谢，但作用机制不同：黄芩素的作用与影响关键酶活性有关，而汉黄芩素的作用与抑制能量代谢关键酶表达有关。 |
| ABSTRACT： | OBJECTIVE To explore the difference in th e mechanis m of baicalein and wogonin inhibiting the energy metabolism of hepatoma cells. METHODS Human hepatoma HepG 2 cells were divided into blank control group （without medicine），different dose groups of baicalein and wogonin （1.25，2.5，5，10 and 20 μmol/L）. The effects of baicalein and wogonin on the viability of HepG 2 cells were detected by MTT assay. HepG 2 cells were divided into blank control group （without medicine），baicalein group and wogonin group. After administration ，the concentration of ATP in cell was detected by enhanced ATP kit. The levels of cell glycolysis and mitochondrial energy metabolism were evaluated by glycolysis and mitochondrial pressure test kit ；the affinity of baicalein and wogonin with key enzymes of energy metabolism was predicted by molecular docking ，and the key enzymes of energy metabolism with high affinity were screened ；the expression of key enzymes of energy metabolism was detected by Western blot. RESULTS Within the dose range of 2.5-20 μmol/L，the half inhibitory concentrations of baicalein and wogonin were 12.84 and 24.09 μmol/L；baicalein 1.25 μmol/L and wogonin 2.5 μmol/L had no effect on cell viability ，so it was selected as the dosage for subsequent experiments. Compared with blank control group ，the concentration of ATP in HepG 2 cells decreased significantly in baicalein group and wogonin group （P＜0.05）；the inhibitory effects on basic acidification rate of HepG 2 cells in wogonin group were significantly stronger than those of baicalein group （P＜0.05），but there was no significant difference between them on the basic oxygen consumption rate （P＞0.05）；baicalein had strong binding to pyruvate kinase M 2 and mitochondrial enzyme complexes Ⅰ（CⅠ），C Ⅱ and C Ⅳ，while wogonin only had strong binding to pyruvate kinase M 2； wogonin could significantly down-regulate the protein expressions of hexokinase ，phosphofructokinase，pyruvate kinase M 2，CⅠ， C Ⅱ and C Ⅳ（P＜0.05），but there was no statistical significance in the effect of baicalein on the regulation of these enzymes （P＞ 0.05）. CONCLUSIONS Both baicalein and wogonin can inhibit the energy metabolism of hepatoma HepG 2 cells，but the mechanism is different ：the effect of baicalein is related to the activity of key enzymes ，while the effect of wogonin is related to the inhibition of the expression of key enzymes of energy metabolism. |
| 期刊： | 2022年第33卷第11期 |
| 作者： | 许珂嘉，张子蒙，付传奎，陈志鹏，李伟东，吴丽 |
| AUTHORS： | XU Kejia，ZHANG Zimeng ，FU Chuankui ，CHEN Zhipeng ，LI Weidong ，WU Li |
| 关键字： | 黄芩素；汉黄芩素；肝癌细胞；能量代谢；糖酵解代谢表型；线粒体能量代谢 |
| KEYWORDS： | baicalein；wogonin；hepatoma cells ；energy |
| 阅读数： | 466 次 |
| 本月下载数： | 11 次 |
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