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大黄素甲醚8-O-β-吡喃葡萄糖苷对皮肤黑色素瘤A375细胞凋亡的影响及机制研究
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| 篇名： | 大黄素甲醚8-O-β-吡喃葡萄糖苷对皮肤黑色素瘤A375细胞凋亡的影响及机制研究 |
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| 摘要： | 目的：研究天然产物大黄素甲醚8-O-β-吡喃葡萄糖苷（PG）对皮肤黑色素瘤A375细胞凋亡的影响及机制。方法：以0、10、20、50 μg/mL的PG作用于A375细胞24、48、72 h后，采用CCK-8法测定细胞的存活率；以0（对照）、20、50 μg/mL的PG作用于A375细胞48 h后，通过流式细胞仪以膜联蛋白Ⅴ/碘化丙啶（PI）双染法检测细胞凋亡率；免疫印迹法检测半胱氨酸天冬氨酸蛋白酶3（Caspase-3）和多腺苷二磷酸核糖聚合酶（PARP）的蛋白表达以及细胞色素C在线粒体内外的蛋白表达；以0（对照）、5、10 μmol/L的PG作用于A375细胞48 h后，采用酶底物法测定细胞中Caspase-8、Caspase-9的活性。结果：PG能有效降低A375细胞的存活率；与对照比较，20、50 μg/mL的PG作用后细胞凋亡率明显升高（P＜0.01），细胞中Caspase-3、PARP及细胞基质中细胞色素C的蛋白表达均明显增强（P＜0.05或P＜0.01），线粒体中细胞色素C蛋白表达明显减弱（P＜0.05或P＜0.01）；5、10 μmol/L的PG作用后细胞中Caspase-9活性明显增强（P＜0.05或P＜0.01），Caspase-8活性无明显变化。结论：PG能抑制A375细胞活性、促进细胞凋亡；其是通过破坏线粒体膜电位、促进细胞色素C外流来发挥促凋亡作用的。 |
| ABSTRACT： | OBJECTIVE： To study the effect and mechanism of physcion 8-O-β-glucopyranoside （PG） on the apoptosis of skin melanoma A375 cells. METHODS： After A375 cells were treated by PG with 0， 10， 20， 50 μg/mL for 24， 48， 72 h， CCK-8 method was adopted to determine the survival rate of cells. After A375 cells were treated by PG with 0 （control）， 20， 50 μg/mL for 48 h， flow cytometry was used to detect the apoptosis rate of cells with membrane protein Ⅴ/propidium iodide （PI） double staining. Immunoblotting was used to detect the protein expressions of Caspase-3 and polyadenyl adenine diphosphate ribose polymerase （PARP） and protein expressions of cytochrome C inside and outside mitochondria. After A375 cells were treated by PG with 0 （control）， 5， 10 μmol/L for 48 h， enzyme substrate method was used to determine the activities of Caspase-8 and Caspase-9. RESULTS： PG can effectively decrease the survival rate of A375 cells. Compared with control， apoptosis rate of cells was obviously increased after treated by PG with 20， 50 μg/mL （P＜0.01）； protein expressions of Caspase-3， PARP in cells and cytochrome C in cell matrix were obviously enhanced （P＜0.05 or P＜0.01）； and protein expression of cytochrome C in mitochondria was obviously weakened （P＜0.05 or P＜0.01）. Caspase-9 activity in cells was obviously enhanced after treated by PG with 5， 10 μmol/L （P＜0.05 or P＜0.01）； and Caspase-8 activity had no obvious changes. CONCLUSIONS： PG can inhibit activity of A375 cells and promote its apoptosis， and its pro-apoptotic effects is achieved by destructing mitochondrial membrane potential and promoting cytochrome C outflow. |
| 期刊： | 2017年第28卷第28期 |
| 作者： | 李晖，李文静，马荣，曹建华，韩志武 |
| AUTHORS： | LI Hui，LI Wenjing，MA Rong，CAO Jianhua，HAN Zhiwu |
| 关键字： | 大黄素甲醚8-O-β-吡喃葡萄糖苷；皮肤黑色素瘤A375细胞；细胞凋亡；线粒体途径 |
| KEYWORDS： | Physcion 8-O-β-glucopyranoside； Skin melanoma A375 cells； Cell apoptosis； Mitochondrial pathway |
| 阅读数： | 317 次 |
| 本月下载数： | 5 次 |
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