[關鍵詞]
[摘要]
目的 觀察注射用益氣復脈(凍干)(YQFM)與順鉑(Cisplatin,DDP)聯(lián)合對肺癌細胞株A549增殖抑制的協(xié)同效應及作用機制。方法 采用CCK-8法檢測YQFM、DDP以及兩藥聯(lián)用對肺癌細胞株A549的增殖影響,并應用兩藥相互作用指數(shù)評價藥物的聯(lián)合效應;采用熒光探針JC-1來檢測線粒體膜電位的變化,Wester Blotting檢測凋亡相關蛋白Bax、Cleaved-Caspase 3表達。結果 藥物作用時間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)組細胞增殖抑制率顯著增加(P<0.05)。與對照組比較,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表達顯著增加(P<0.05);DDP質量濃度為5.0、10.0 μg/mL時,聯(lián)合用藥組與DDP單獨給藥組比較,Bax蛋白表達量顯著升高(P<0.05)。與對照組比較,各給藥組凋亡蛋白Cleaved-caspase 3表達量顯著升高(P<0.05);DDP質量濃度為5.0、10.0 μg/mL時,聯(lián)合用藥組與DDP單獨給藥組比較,Cleaved-caspase 3蛋白表達量顯著升高(P<0.05)。結論 YQFM聯(lián)合DDP對A549細胞增殖存在協(xié)同抑制作用,機制可能與誘導線粒體凋亡相關。
[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]
[基金項目]
天津市中藥注射劑關鍵技術校企協(xié)同創(chuàng)新實驗室建設項目(17PTSYJC00090)