首页|过氧麦角甾醇通过调控线粒体凋亡途径诱导人肝癌细胞凋亡

过氧麦角甾醇通过调控线粒体凋亡途径诱导人肝癌细胞凋亡

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探讨过氧麦角甾醇(ergosterol peroxide，EP)对人肝癌细胞凋亡的影响及其作用机制。利用cell counting kit-8(CCK-8)法检测 0(空白对照)、2。5、5、10、20、40、80 μmol·L-1的EP作用于HepG2、SK-Hep-1 细胞 24、48、72 h后的细胞活力，并计算24、48、72 h的半数抑制浓度(IC50)。正式实验以 0(空白对照)、10、20、40 μmol·L-1的EP与HepG2 细胞共培养 48 h后，使用活性氧(reactive oxygen species，ROS)荧光探针法检测EP对细胞内ROS的影响，使用线粒体膜电位荧光探针染色检测EP对细胞内线粒体膜电位的影响，使用Annexin V-FITC/PI双染法检测细胞凋亡数，使用AO/EB染色法检测不同浓度的EP对细胞凋亡形态的影响。使用蛋白免疫印迹(Western blot)法检测不同浓度的EP 对线粒体凋亡途径相关蛋白B细胞淋巴瘤 2(Bcl-2)、细胞色素C(Cyt-C)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶 3(caspase-3)、活化的胱天蛋白酶 3(cleaved caspase-3)、胱天蛋白酶 9(caspase-9)、活化的胱天蛋白酶 9(cleaved caspase-9)蛋白表达的影响。结果表明，不同浓度的EP均可抑制肝癌细胞增殖，且呈浓度和时间依赖性。与空白对照组相比，EP药物处理组ROS水平显著升高(P<0。05)，线粒体膜电位显著下降(P<0。05)，细胞总凋亡率显著升高(P<0。05)，Bcl-2蛋白表达显著下调，Cyt-C、Bax、cleaved caspase-9、cleaved caspase-3 表达均显著上调(P<0。05)。综上所述，EP可能通过调控线粒体介导的凋亡途径抑制肝癌细胞增殖，并诱导其凋亡。
Ergosterol peroxide inducing apoptosis of human hepatocellular carcinoma by regulating mitochondrial apoptosis pathway
This study aims to investigate the effect of ergosterol peroxide(EP)on the apoptosis of human hepatocellular carcinoma and its mechanism of action.The cell viability of HepG2 and SK-Hep-1 cells with 0(blank control),2.5,5,10,20,40,and 80 μmol·L-1 of EP after 24,48,and 72 h of action was detected by using CCK-8 assay,and the half inhibitory concentrations(IC50)at 24,48,and 72 h were calculated.Formal experiments were performed to detect the effect of EP on intracellular reactive oxygen species(ROS)using DCFH-DA staining,the effect of EP on intracellular mitochondrial membrane potential using JC-1 staining,the number of apoptotic cells using Annexin V-FITC/PI double-staining after HepG2 cells were co-cultured with 0(blank control),10,20,40 μmol·L-1 EP for 48 h.The effects of EP at different concentrations on apoptotic morphology were detected using AO/EB staining.The effects of different concentrations of EP on the protein expression of mitochondrial apoptosis pathway-related proteins B cell lymphoma 2(Bcl-2),cytochrome C(Cyt-C),Bcl-2-related X protein(Bax),caspase-3,cleaved caspase-3,caspase-9,and cleaved caspase-9 were examined by using Western blot.The results showed that different concentrations of EP could inhibit the proliferation of hepatocellular carcinoma with concentration-and time-dependent trends.Compared with the blank control group,the ROS level in the EP-treated group increased significantly(P<0.05).The mitochondrial membrane potential decreased significantly(P<0.05).The total apoptosis rate increased significantly(P<0.05).The expression of Bcl-2 protein was significantly down-regulated,and the expression of Cyt-C,Bax,cleaved caspase-9,and cleaved caspase-3 were significantly up-regulated(P<0.05).In summary,EP may inhibit the proliferation of hepatocellular carcinoma by modulating the mitochondria-mediated apoptosis pathway and induce apoptosis.

ergosterol peroxidehepatocellular carcinomamitochondrial membrane potentialmitochondrial apoptosisreactive ox-ygen species

王璐、罗然、赵音旭、邹宇、卜明、林宇

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齐齐哈尔医学院 药学院,黑龙江 齐齐哈尔 161006

过氧麦角甾醇 肝癌 线粒体膜电位 线粒体凋亡 活性氧

黑龙江省自然科学基金联合引导项目齐齐哈尔市科技局联合引导项目齐齐哈尔市科技局联合引导项目

LH2022H113LHYD2021005LHYD2021021

2024

10.19540/j.cnki.cjcmm.20240221.501

中国中药杂志
中国药学会

中国中药杂志

CSTPCD北大核心
影响因子：1.718
ISSN：1001-5302
年,卷(期)：2024.49(12)
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