低聚葡萄籽原花青素联合顺铂对人肺腺癌细胞A549凋亡的影响
发布时间：2018-12-23 19:34
【摘要】：目的:研究低聚葡萄籽原花青素(oligomeric?grape?seed?proanthocyanidins,O-GSP)联合顺铂对人肺腺癌细胞A549抗氧化、细胞凋亡及凋亡相关蛋白表达的影响。方法:细胞分为对照组、顺铂(5 mg/L)组、O-GSP(4 mg/L)组、O-GSP(4 mg/L)+顺铂(5 mg/L)组。硫代巴比妥酸法测定细胞中丙二醛(malondialdehyde,MDA)含量;二硫代二硝基苯甲酸法测定谷胱甘肽(glutathione,GSH)含量;黄嘌呤氧化酶法测定超氧化物歧化酶(superoxide?dismutase,SOD)活力;流式细胞术检测细胞内活性氧(reactive?oxygen?species,ROS)含量和细胞凋亡率;Western?blot检测细胞凋亡相关蛋白Bcl-2、Bax和caspase-3的表达。结果:顺铂导致A549细胞MDA、ROS含量显著升高(P0.05),GSH含量和SOD活力显著降低(P0.05),O-GSP对细胞内MDA含量、GSH含量、SOD活力无显著影响(P0.05)。O-GSP+顺铂组相对于顺铂组,细胞内MDA、GSH、SOD水平无显著性变化(P0.05),而ROS含量明显降低(P0.05)。顺铂、O-GSP以及两者联用均能显著促进细胞凋亡(P0.05),并且能够显著提高Bax及Bax/Bcl-2值,顺铂和O-GSP+顺铂可显著降低Bcl-2的表达(P0.05),同时激活caspase-3。结论:O-GSP联合顺铂可促进A549细胞的凋亡,其机制与O-GSP的抗氧化作用没有显著联系,而是与两者可共同调节凋亡相关蛋白基因表达有关。
[Abstract]:Aim: to study the effects of oligograpetseed proanthocyanidins (oligomeric?grape?seed?proanthocyanidins,O-GSP) combined with cisplatin on antioxidation, apoptosis and expression of apoptosis-related proteins in human lung adenocarcinoma cell line A549. Methods: the cells were divided into control group, cisplatin (5 mg/L) group, O-GSP (4 mg/L) group and O-GSP (4 mg/L) cisplatin (5 mg/L) group. The contents of malondialdehyde (malondialdehyde,MDA), glutathione (glutathione,GSH) and superoxide dismutase (superoxide?dismutase,SOD) in cells were determined by thiobarbituric acid method, dinitrobenzoic acid method and xanthine oxidase method respectively. Flow cytometry was used to detect the content of reactive oxygen species (reactive?oxygen?species,ROS) and apoptosis rate, and Western?blot was used to detect the expression of Bcl-2,Bax and caspase-3. Results: Cisplatin significantly increased the MDA,ROS content (P0.05), GSH content and SOD activity decreased significantly (P0.05), O-GSP induced MDA content and GSH content in A549 cells. There was no significant effect on SOD activity (P0.05). Compared with cisplatin group, the intracellular MDA,GSH,SOD level of O-GSP cisplatin group had no significant change (P0.05), but the content of ROS decreased significantly (P0.05). Cisplatin, O-GSP and their combination significantly promoted apoptosis (P0.05), and significantly increased Bax and Bax/Bcl-2 values. Cisplatin and O-GSP cisplatin significantly decreased the expression of Bcl-2 (P0.05). Simultaneous activation of caspase-3. Conclusion: O-GSP combined with cisplatin can promote the apoptosis of A549 cells, and its mechanism is not related to the antioxidant effect of O-GSP, but to the regulation of apoptosis-related protein gene expression by both of them.
【作者单位】： 北京联合大学应用文理学院生物活性物质与功能食品北京市重点实验室;
【基金】：北京市自然科学基金项目(7163211)
【分类号】：R734.2
[Abstract]:Aim: to study the effects of oligograpetseed proanthocyanidins (oligomeric?grape?seed?proanthocyanidins,O-GSP) combined with cisplatin on antioxidation, apoptosis and expression of apoptosis-related proteins in human lung adenocarcinoma cell line A549. Methods: the cells were divided into control group, cisplatin (5 mg/L) group, O-GSP (4 mg/L) group and O-GSP (4 mg/L) cisplatin (5 mg/L) group. The contents of malondialdehyde (malondialdehyde,MDA), glutathione (glutathione,GSH) and superoxide dismutase (superoxide?dismutase,SOD) in cells were determined by thiobarbituric acid method, dinitrobenzoic acid method and xanthine oxidase method respectively. Flow cytometry was used to detect the content of reactive oxygen species (reactive?oxygen?species,ROS) and apoptosis rate, and Western?blot was used to detect the expression of Bcl-2,Bax and caspase-3. Results: Cisplatin significantly increased the MDA,ROS content (P0.05), GSH content and SOD activity decreased significantly (P0.05), O-GSP induced MDA content and GSH content in A549 cells. There was no significant effect on SOD activity (P0.05). Compared with cisplatin group, the intracellular MDA,GSH,SOD level of O-GSP cisplatin group had no significant change (P0.05), but the content of ROS decreased significantly (P0.05). Cisplatin, O-GSP and their combination significantly promoted apoptosis (P0.05), and significantly increased Bax and Bax/Bcl-2 values. Cisplatin and O-GSP cisplatin significantly decreased the expression of Bcl-2 (P0.05). Simultaneous activation of caspase-3. Conclusion: O-GSP combined with cisplatin can promote the apoptosis of A549 cells, and its mechanism is not related to the antioxidant effect of O-GSP, but to the regulation of apoptosis-related protein gene expression by both of them.
【作者单位】： 北京联合大学应用文理学院生物活性物质与功能食品北京市重点实验室;
【基金】：北京市自然科学基金项目(7163211)
【分类号】：R734.2
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