摘要
目的 探讨丹参多糖经丝裂原激活蛋白激酶(MAPK)/细胞外信号调节激酶(ERK)信号轴抑制对肺癌细胞A549增殖、迁移和凋亡的影响。方法 体外培养A549细胞,用不同质量浓度(0,1,2,4,8,16,32,64 mg/mL)丹参多糖干预48 h后,确定丹参多糖的半数抑制浓度(IC50);将A549细胞分为对照组、高剂量组、中剂量组和低剂量组(分别以0,8,4,2 mg/mL丹参多糖干预),以及抑制剂组(40μmol/L PD 98059)。采用划痕愈合试验检测A549细胞的迁移水平,采用流式细胞术检测A549细胞的凋亡情况,同时采用实时荧光定量聚合酶链反应法和免疫印迹法分别检测A549细胞中MAPK、ERK、基质金属蛋白酶-9(MMP-9)、B细胞淋巴瘤2基因(Bcl-2)、胱天蛋白酶3(caspase-3)mRNA和蛋白表达水平。结果 随着丹参多糖质量浓度的增加,A549细胞增殖率显著降低(P <0.05);丹参多糖对A549细胞的IC50为7.82 mg/mL,高、中、低剂量组干预剂量分别为8,4,2 mg/mL。与对照组比较,抑制剂组,高、中、低剂量组细胞的迁移距离及MMP-9,MAPK,ERK,Bcl-2 mRNA和蛋白表达水平均显著降低(P <0.05),细胞凋亡率、caspase-3 mRNA和蛋白表达水平均显著升高(P <0.05);与抑制剂组比较,高、中、低剂量组细胞的迁移距离及MMP-9,MAPK,ERK,Bcl-2 mRNA和蛋白表达水平均显著升高(P <0.05),细胞凋亡率、caspase-3 mRNA和蛋白表达水平均显著降低(P <0.05),且随丹参多糖剂量的增加呈量效依赖关系(P <0.05)。结论 丹参多糖可能通过MAPK/ERK信号轴诱导A549细胞凋亡,抑制A549细胞增殖和迁移。
Objective To investigate the effect of Salvia miltiorrhiza polysaccharide on proliferation,migration and apoptosis of lung cancer cells A549 through mitogen active protein kinase(MAPK)/extracellular signal-regulated kinase(ERK)signal axis inhibition.Methods A549 cells were cultured in vitro and treated with different mass concentrations(0,1,2,4,8,16,32,64 mg/mL)of Salvia miltiorrhiza polysaccharides for 48 h to determine the half inhibitory concentration(IC50)of Salvia miltiorrhiza polysaccharides.A549 cells were divided into the control group,high-dose group,medium-dose group,and low-dose group(0,8,4,2 mg/mL of Salvia miltiorrhiza polysaccharide),and the inhibitor group(40μmol/L PD 98059).The migration of A549 cells was detected by the scratch healing assay,and the apoptosis of A549 cells was detected by the flow cytometry.The mRNA and protein expression levels of matrix metalloproteinase-9(MMP-9),MAPK,ERK,B cell lymphoma-2(Bcl-2)and caspase-3 were detected by the reverse transcription-polymerase chain reaction(RT-PCR)and the Western blot respectively.Results With the increase of the mass concentration of Salvia miltiorrhiza polysaccharides,the proliferation rate of A549 cells significantly decreased(P<0.05).The IC50 of Salvia miltiorrhiza polysaccharides on A549 cells was 7.82 mg/mL,and 8,4,and 2 mg/mL were selected as the intervention doses for high-,medium-,and low-dose groups respectively.Compared with those in the control group,the migration distance of cells,MMP-9,MAPK,ERK,and Bcl-2 mRNA and protein expression levels significantly decreased(P<0.05),while the apoptosis rate and caspase-3 mRNA and protein expression levels significantly increased in the inhibitor group and the high-,medium-,and low-dose groups(P<0.05).Compared with those in the inhibitor group,the migration distance of cells,MMP-9,MAPK,ERK,and Bcl-2 mRNA and protein expression levels significantly increased(P<0.05),while the apoptosis rate and caspase-3 mRNA and protein expression levels significantly decreased in the high-,medium-,and low-dose groups(P<0.05),which showed a dose-dependent relationship with the increase of Salvia miltiorrhiza polysaccharide dose(P<0.05).Conclusion Salvia miltiorrhiza polysaccharide can induce apoptosis of A549 cells and inhibit their proliferation and migration through the MAPK/ERK signaling axis.
作者
郭志青
支学军
王布
苏菁
刘芳
GUO Zhiqing;ZHI Xuejun;WANG Bu;SU Jing;LIU Fang(The First Affiliated Hospital of Hebei North University,Zhangjiakou,Hebei,China 075000)
出处
《中国药业》
CAS
2024年第1期40-44,共5页
China Pharmaceuticals
基金
河北省中医药管理局2023年度中医药类科学研究课题计划项目[2023331]。
