紫云英苷对肺癌A549细胞增殖、迁移、凋亡和氧化应激的影响

Effect of Astragalin on proliferation,migration,apoptosis and oxidative stress of lung cancer A549 cells

目的 探讨紫云英苷(AG)对肺癌A549细胞增殖、迁移、凋亡和氧化应激的影响及其机制。方法 将体外培养的A549细胞分为AG低(AG-L)、中(AG-M)、高(AG-H)剂量(分别给予25、50、100μmol·L^(-1)AG处理)组和AG-H+核因子E2相关因子2(Nrf2)/抗氧化反应元件(ARE)通路激活剂SFN(AG-H+SFN,100μmol·L^(-1)AG+10μmol·L^(-1)SFN)组及对照组(正常培养不做处理)。用噻唑蓝法、Transwell小室和流式细胞仪分别检测细胞增殖、迁移和凋亡能力,用蛋白质印迹法检测细胞中B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、醌氧化还原酶1(NQO1)和血红素加氧酶-1(HO-1)蛋白的表达水平。结果 对照组和AG-L、AG-M、AG-H组及AG-H+SFN组的细胞增殖活力(48 h时光密度值)分别为0.62±0.04、0.54±0.03、0.43±0.03、0.34±0.03和0.48±0.03,迁移细胞数分别为(121.06±11.79)、(101.17±7.10)、(74.22±5.64)、(37.83±4.24)和(87.06±6.19)个,细胞凋亡率分别为(6.50±1.41)%、(13.78±2.03)%、(25.57±3.37)%、(38.10±3.40)%和(10.13±1.56)%,SOD水平分别为(21.11±1.15)、(18.93±1.03)、(15.79±0.81)、(13.65±0.85)和(17.64±1.14)U·mL^(-1),MDA水平分别为(4.33±0.46)、(6.42±0.61)、(7.21±0.66)、(8.85±0.89)和(6.58±0.60)nmol·mL^(-1),Nrf2蛋白相对表达水平分别为1.26±0.08、1.08±0.08、0.86±0.05、0.61±0.03和0.90±0.05,HO-1蛋白相对表达水平分别为0.85±0.06、0.76±0.05、0.57±0.04、0.23±0.03和0.56±0.04;AG-L、AG-M、AG-H组的上述指标与对照组比较,AG-H+SFN组的上述指标和AG-H组比较,差异均有统计学意义(均P<0.05)。结论 AG可通过抑制Nrf2/ARE通路活化抑制肺癌A549细胞增殖、迁移并诱导细胞凋亡和氧化应激。

Objective To investigate the effects of astragalin(AG)on proliferation,migration,apoptosis and oxidative stress of lung cancer A549 cells and its mechanism.Methods A549 cells cultured in vitro were divided into low,medium and high AG groups(AG-L、AG-M、AG-H,25,50 and 100μmol·L^(-1) AG,respectively)and AG-H+nuclear factor groups.The proliferation,migration and apoptosis of nuclear factor E2-related factor 2(Nrf2)/antioxidant reaction element(ARE)pathway activator SFN(AG-H+SFN,100μmol·L^(-1) AG+10μmol·L^(-1) SFN)group and control group(normal culture without treatment) were detected by methyl thiazolyl tetrazolium method,Transwell chamber and flow cytometry. Theexpression of B - cell lymphoma - 2 ( Bcl - 2 ) ,Bcl - 2 associated X protein ( Bax) ,quinone oxidoredutase 1( NQO1) and heme oxygenase - 1 ( HO - 1) proteins were detected by Western blot. Results The proliferationactivity ( optical density value at 48 h) of the control group,AG - L,AG - M,AG - H and AG - H + SFN groupswere 0. 62 ± 0. 04,0. 54 ± 0. 03,0. 43 ± 0. 03,0. 34 ± 0. 03,0. 48 ± 0. 03;the number of migrating cell swere( 121. 06 ± 11. 79) ,( 101. 17 ± 7. 10) ,( 74. 22 ± 5. 64) ,( 37. 83 ± 4. 24) and ( 87. 06 ± 6. 19);the apoptosis rateswere ( 6. 50 ± 1. 41) %,( 13. 78 ± 2. 03) %,( 25. 57 ± 3. 37) %,( 38. 10 ± 3. 40) %,( 10. 13 ± 1. 56) %;SODlevels were ( 21. 11 ± 1. 15) ,( 18. 93 ± 1. 03) ,( 15. 79 ± 0. 81) ,( 13. 65 ± 0. 85) and ( 17. 64 ± 1. 14) U·mL^(-1);MDA levels were ( 4. 33 ± 0. 46) ,( 6. 42 ± 0. 61) ,( 7. 21 ± 0. 66) ,( 8. 85 ± 0. 89) ,( 6. 58 ± 0. 60) nmol·mL^(-1);Nrf2 protein expression levels were 1. 26 ± 0. 08,1. 08 ± 0. 08,0. 86 ± 0. 05,0. 61 ± 0. 03,0. 90 ± 0. 05;HO - 1protein expression levels were 0. 85 ± 0. 06,0. 76 ± 0. 05,0. 57 ± 0. 04,0. 23 ± 0. 03,0. 56 ± 0. 04,respectively.There were statistically significant differences in the above indicators between AG - L,AG - M,AG - H groups andcontrol group,between AG - H + SFN group and AG - H group ( all P < 0. 05) . Conclusion AG can inhibit theproliferation and migration of lung cancer A549 cells and induce apoptosis and oxidative stress by inhibiting theactivation of Nrf2 /ARE pathway.