萝藦果壳多糖诱导人肺癌A549细胞凋亡的研究  

Induction of apoptosis in human lung cancer A549 cells by polysaccharides from Metaplexis japonica Makino nutshell

作　　者：李佳楠 韩冠英 郭斌 LI Jia-nan;HAN Guan-ying;GUO Bin(School of Pharmaceutical Sciences,Jinzhou Medical University,Jinzhou,Liaoning 121000,China;不详)

机构地区：[1]锦州医科大学药学院,辽宁锦州121000 [2]锦州医科大学附属第一医院,辽宁锦州121000

出　　处：《现代预防医学》2022年第2期339-343,共5页Modern Preventive Medicine

基　　金：辽宁省“兴辽英才计划”项目(XLYC1907187)。

摘　　要：目的探究萝藦果壳多糖(MJP-1’)诱导人肺癌A549细胞凋亡的作用。方法采用CCK8法检测萝藦果壳多糖对A549细胞活力的影响;采用比色法检测A549细胞内乳酸脱氢酶的释放量变化情况;通过TUNEL实验探究萝藦果壳多糖对A549细胞凋亡的影响;通过JC-1荧光探针探究萝藦果壳多糖对A549细胞线粒体膜电位的影响;通过蛋白质分子印迹法探究萝藦果壳多糖作用后A549细胞凋亡相关蛋白表达的变化。结果 CCK8法显示萝藦果壳多糖抑制A549细胞的增殖,并呈剂量依赖趋势(125μg/ml组:t=3.092,P=0.015;250μg/ml组:t=3.929,P=0.004;500μg/ml组:t=5.626,P<0.001;1 000μg/ml组:t=7.512,P<0.001;2 000μg/ml组:t=9.677,P<0.001;4 000μg/ml组:t=12.290,P<0.001;8 000μg/ml组:t=16.000,P<0.001);比色法显示A549细胞乳酸脱氢酶释放量增加(2 mg/ml组:t=8.233,P=0.001;4 mg/ml组:t=12.640,P<0.001;8 mg/ml组:t=27.530,P<0.001);TUNEL实验显示萝藦果壳多糖可促进A549细胞凋亡(2 mg/ml组:t=7.803,P<0.001;4 mg/ml组:t=12.460,P<0.001;8 mg/ml组:t=39.340,P<0.001);JC-1探针实验结果显示萝藦果壳多糖导致A549细胞线粒体膜电位降低(2 mg/ml组:t=24.120,P<0.001;4 mg/ml组:t=28.530,P<0.001;8 mg/ml组:t=31.600,P<0.001);蛋白质分子印迹法结果显示萝藦果壳多糖可以下调Bcl-2蛋白表达(2 mg/ml组:t=3.790,P=0.019;4 mg/ml组:t=6.598,P=0.003;8 mg/ml组:t=12.840,P<0.001),并使Bax、cleavedcaspase-3蛋白表达增加(Bax组:2 mg/ml组:t=17.280,P<0.001;4 mg/ml组:t=36.040,P<0.001;8 mg/ml组:t=22.520,P<0.001;cleaved-caspase-3组:2 mg/ml组:t=6.487,P=0.003;4 mg/ml组:t=9.326,P<0.001;8 mg/ml组:t=22.380,P<0.001)。结论本研究发现萝藦果壳多糖可以诱导人肺癌A549细胞凋亡,为进一步开发利用萝藦果壳多糖奠定了基础。Objective To Explore the role of polysaccharides from Metaplexis japonica Makino nutshell(MJP-1’) in inducing apoptosis of human lung cancer A549 cells.Methods The CCK8 method was used to detect the effect of MJP-1’ on the viability of A549 cells.Colorimetric method was used to detect the changes in the release of lactate dehydrogenase in A549 cells.The TUNEL experiment was used to explore the effects of MJP-1’ on A549 cell apoptosis.The effect of MJP-1’ on the mitochondrial membrane potential of A549 cells were explored with JC-1 fluorescent probe.The expression of apoptosis-re-lated proteins in A549 cells after the action of MJP-1’ was explored by Western Blot experiment.Results The CCK8 method showed that MJP-1’ could inhibit the proliferation of A549 cells in a dose-dependent manner(125 μg/ml:t=3.092,P=0.015;250 μg/ml:t=3.929,P=0.004;500 μg/ml:t =5.626,P <0.001;1 000 μg/ml:t =7.512,P <0.001;2 000 μg/ml:t =9.677,P <0.001;4 000 μg/ml:t=12.290,P<0.001;8 000 μg/ml:t=16.000,P<0.001).Colorimetry showed that the release of lactate dehydrogenase was increased in A549 cells(2 mg/ml:t=8.233,P=0.001;4 mg/ml:t=12.640,P<0.001;8 mg/ml:t=27.530,P<0.001).TUNEL experiment showed that MJP-1’ could promote A549 cell apoptosis(2 mg/ml:t=7.803,P<0.001;4 mg/ml:t=12.460,P<0.001;8 mg/ml:t=39.340,P<0.001).JC-1 probe experiment showed that MJP-1’ caused a decrease in the mitochondrial membrane potential of A549 cells(2 mg/ml:t=24.120,P<0.001;4 mg/ml:t=28.530,P<0.001;8 mg/ml:t=31.600,P<0.001).The Western Blot results showed that MJP-1’ could down-regulate the expression of Bcl-2 protein and increase the expression of Bax and cleaved-caspase-3 proteins(Bcl-2:2 mg/ml:t=3.790,P=0.019;4 mg/ml:t=6.598,P=0.003;8 mg/ml:t =12.840,P <0.001;Bax:2 mg/ml:t =17.280,P <0.001;4 mg/ml:t =36.040,P <0.001;8 mg/ml:t =22.520,P <0.001;cleaved-caspase-3:2 mg/ml:t=6.487,P=0.003;4 mg/ml:t=9.326,P<0.001;8 mg/ml:t=22.380,P<0.001).Conclusion MJP-1’ can induce apoptosis of human lung cancer A549 cells,which lays a foundation for develop

关 键 词：萝藦果壳 多糖 人肺癌A549细胞 细胞凋亡 

分 类 号：R285[医药卫生—中药学]