[關(guān)鍵詞]
[摘要]
目的 觀察注射用益氣復(fù)脈(凍干)(YQFM)與順鉑(Cisplatin,DDP)聯(lián)合對(duì)肺癌細(xì)胞株A549增殖抑制的協(xié)同效應(yīng)及作用機(jī)制。方法 采用CCK-8法檢測(cè)YQFM、DDP以及兩藥聯(lián)用對(duì)肺癌細(xì)胞株A549的增殖影響,并應(yīng)用兩藥相互作用指數(shù)評(píng)價(jià)藥物的聯(lián)合效應(yīng);采用熒光探針JC-1來檢測(cè)線粒體膜電位的變化,Wester Blotting檢測(cè)凋亡相關(guān)蛋白Bax、Cleaved-Caspase 3表達(dá)。結(jié)果 藥物作用時(shí)間48 h,與DDP等劑量組比較,聯(lián)合用藥(YQFM 16.25 mg/mL+DDP 1.25 μg/mL、YQFM 16.25 mg/mL+DDP 2.5 μg/mL、YQFM 16.25 mg/mL+DDP 5.00 μg/mL)組細(xì)胞增殖抑制率顯著增加(P<0.05)。與對(duì)照組比較,各給藥組凋亡率均顯著升高(P<0.05);與DDP組比較,聯(lián)合用藥組凋亡率顯著升高(P<0.05)。與對(duì)照組比較,DDP 5.0、10.0 μg/mL組和各聯(lián)合用藥組促凋亡蛋白Bax表達(dá)顯著增加(P<0.05);DDP質(zhì)量濃度為5.0、10.0 μg/mL時(shí),聯(lián)合用藥組與DDP單獨(dú)給藥組比較,Bax蛋白表達(dá)量顯著升高(P<0.05)。與對(duì)照組比較,各給藥組凋亡蛋白Cleaved-caspase 3表達(dá)量顯著升高(P<0.05);DDP質(zhì)量濃度為5.0、10.0 μg/mL時(shí),聯(lián)合用藥組與DDP單獨(dú)給藥組比較,Cleaved-caspase 3蛋白表達(dá)量顯著升高(P<0.05)。結(jié)論 YQFM聯(lián)合DDP對(duì)A549細(xì)胞增殖存在協(xié)同抑制作用,機(jī)制可能與誘導(dǎo)線粒體凋亡相關(guān)。
[Key word]
[Abstract]
objective To observe the synergistic effect and mechanism of combined injection of Yiqi Fumai Lyophilized Injection (YQFM) and Cisplatin (DDP) on the proliferation inhibition of A549 lung cancer cell line. Methods The CCK-8 method was used to detect the effect of single DDP and YQFM, and combination of two drugs on the proliferation of lung cancer cell line A549. The changes of mitochondrial membrane potential were detected by fluorescent probe JC-1, which labeled the mitochondrial membrane potential. Western Blot was used to detect the expression of apoptosis-related proteins Bax and Caspase3. Results Compared with DDP equal dose group, the inhibition rate of cell proliferation was significantly increased in YQFM 16.25 mg/mL + DDP 1.25 μg/mL, YQFM 16.25 mg/mL + DDP 2.5 μg/mL, YQFM 16.25 mg/mL + DDP 5.00 μg/mL groups (P<0.05). Compared with the control group, the apoptotic rate of each drug group was significantly higher (P<0.05), and the apoptotic rate of the combined drug group was significantly higher (P<0.05) than that of the DDP group. Compared with control group, the expression of apoptotic protein Bax in DDP 5.0, 10.0 μg/mL and combination groups was significantly higher (P<0.05); when DDP concentration was 5.0 and 10.0 μg/mL, the expression of Bax protein in combination group was significantly higher than that in DDP alone group (P<0.05). Compared with the control group, the expression of apoptotic protein Cleaved-caspase 3 was significantly increased in each group (P<0.05). When the concentration of DDP was 5.0 and 10.0 μg/mL, the expression of Cleaved-caspase 3 in the combined group was significantly higher than that in the DDP alone group (P<0.05). Conclusion YQFM combined with DDP could inhibit the proliferation of A549 cells synergistically, and the mechanism might be related to the induction of mitochondrial apoptosis.
[中圖分類號(hào)]
[基金項(xiàng)目]
天津市中藥注射劑關(guān)鍵技術(shù)校企協(xié)同創(chuàng)新實(shí)驗(yàn)室建設(shè)項(xiàng)目(17PTSYJC00090)