【摘要】：目的:研究低聚葡萄籽原花青素(oligomeric?grape?seed?proanthocyanidins,O-GSP)聯(lián)合順鉑對人肺腺癌細胞A549抗氧化、細胞凋亡及凋亡相關蛋白表達的影響。方法:細胞分為對照組、順鉑(5 mg/L)組、O-GSP(4 mg/L)組、O-GSP(4 mg/L)+順鉑(5 mg/L)組。硫代巴比妥酸法測定細胞中丙二醛(malondialdehyde,MDA)含量;二硫代二硝基苯甲酸法測定谷胱甘肽(glutathione,GSH)含量;黃嘌呤氧化酶法測定超氧化物歧化酶(superoxide?dismutase,SOD)活力;流式細胞術檢測細胞內活性氧(reactive?oxygen?species,ROS)含量和細胞凋亡率;Western?blot檢測細胞凋亡相關蛋白Bcl-2、Bax和caspase-3的表達。結果:順鉑導致A549細胞MDA、ROS含量顯著升高(P0.05),GSH含量和SOD活力顯著降低(P0.05),O-GSP對細胞內MDA含量、GSH含量、SOD活力無顯著影響(P0.05)。O-GSP+順鉑組相對于順鉑組,細胞內MDA、GSH、SOD水平無顯著性變化(P0.05),而ROS含量明顯降低(P0.05)。順鉑、O-GSP以及兩者聯(lián)用均能顯著促進細胞凋亡(P0.05),并且能夠顯著提高Bax及Bax/Bcl-2值,順鉑和O-GSP+順鉑可顯著降低Bcl-2的表達(P0.05),同時激活caspase-3。結論:O-GSP聯(lián)合順鉑可促進A549細胞的凋亡,其機制與O-GSP的抗氧化作用沒有顯著聯(lián)系,而是與兩者可共同調節(jié)凋亡相關蛋白基因表達有關。
[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.
【作者單位】： 北京聯(lián)合大學應用文理學院生物活性物質與功能食品北京市重點實驗室;
【基金】：北京市自然科學基金項目(7163211)
【分類號】：R734.2

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