目的 探究羽扇豆醇-3β-琥珀酸酯對HepG2細胞凋亡的作用及機制。方法 采用0、2.5、5、10、20、40、80 μmol/L的羽扇豆醇-3β-琥珀酸酯作用于HepG2細胞24、48、72 h，采用CCK-8法檢測各組細胞的增殖活性。采用0、4、8、16 μmol/L的羽扇豆醇-3β-琥珀酸酯作用于HepG2細胞48 h后，AO/EB染色法和Annexin V-FITC/PI雙染法檢測各組細胞的凋亡率；JC-1染色法觀察HepG2細胞的線粒體膜電位變化；DCFH-DA染色法觀察細胞的活性氧（ROS）水平。Western blotting法檢測各組細胞線粒體凋亡途徑相關(guān)蛋白表達變化。結(jié)果 羽扇豆醇-3β-琥珀酸酯呈時間和濃度相關(guān)性地抑制HepG2細胞增殖（P＜0.01）。與對照組相比，羽扇豆醇-3β-琥珀酸酯組細胞的ROS水平、總凋亡率、Bcl-2相關(guān)X蛋白（Bax）、細胞色素C、裂解半胱氨酸蛋白酶9（cleaved-Caspase-9）、cleaved Caspase-3蛋白水平呈濃度相關(guān)性升高（P＜0.01、0.001）；線粒體的膜電位、Bcl-2蛋白表達則顯著降低（P＜0.01）。結(jié)論 羽扇豆醇-3β-琥珀酸酯可能通過調(diào)控線粒體凋亡途徑誘導(dǎo)人肝癌HepG2細胞凋亡，進而發(fā)揮抗肝癌作用。

Objective To investigate the effect of lupinol-3β-succinate on apoptosis of HepG2 cells and mechanism. Methods HepG2 cells were treated with lupinol-3β-succinate (0, 2.5, 5, 10, 20, 40 and 80 μmol/L) for 24, 48, and 72 h, and the proliferation activity of the cells was detected by CCK-8 method. HepG2 cells were treated with lupinol-3β-succinate (0, 4, 8, and 16 μmol/L) for 48 h, and the apoptosis rate of each group was detected by AO/EB staining and Annexin V-FITC/PI double staining. Changes of mitochondrial membrane potential in HepG2 cells were observed by JC-1 staining. Levels of ROS were observed by DCFH-DA staining. Western blotting method was used to detect the changes of mitochondrial apoptosis-related protein expression in each group. Results Lupinol-3β-succinate showed significant time and dose dependent inhibition of HepG2 cell proliferation (P < 0.01). Compared with control group, ROS level, total apoptosis rate, Bax, Cyt-C, cleaved Caspase-9, and cleaved Caspase-3 protein levels were increased in a dose-dependent manner after lupinol-3β-succinate treatment (P < 0.01, 0.001). Mitochondrial membrane potential and Bcl-2 protein expression were significantly decreased (P < 0.01). Conclusion Lupinol-3β-succinate may induce apoptosis of human hepatoma HepG2 cells by regulating mitochondrial apoptosis pathway, and finally play an anti-hepatocellular carcinoma role.

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中央支持地方高校改革發(fā)展資金人才培養(yǎng)項目（21B024）；齊齊哈爾醫(yī)學(xué)科學(xué)院科研項目（QMSI2022Z-01）