【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(xì)胞A549抗氧化、細(xì)胞凋亡及凋亡相關(guān)蛋白表達(dá)的影響。方法:細(xì)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(xì)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(xì)胞術(shù)檢測細(xì)胞內(nèi)活性氧(reactive?oxygen?species,ROS)含量和細(xì)胞凋亡率;Western?blot檢測細(xì)胞凋亡相關(guān)蛋白Bcl-2、Bax和caspase-3的表達(dá)。結(jié)果:順鉑導(dǎo)致A549細(xì)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(xì)胞內(nèi)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(xì)胞內(nèi)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(jìn)細(xì)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(dá)(P0.05),同時激活caspase-3。結(jié)論:O-GSP聯(lián)合順鉑可促進(jìn)A549細(xì)胞的凋亡,其機(jī)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(diào)節(jié)凋亡相關(guān)蛋白基因表達(dá)有關(guān)。
[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(xué)應(yīng)用文理學(xué)院生物活性物質(zhì)與功能食品北京市重點(diǎn)實(shí)驗(yàn)室;
【基金】：北京市自然科學(xué)基金項(xiàng)目(7163211)
【分類號】：R734.2

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