目的 探讨甘草查尔酮A(LCA)对胶质瘤U87细胞增殖、迁移、侵袭和抗氧化能力的影响及其机制.方法 将体外培养的胶质瘤U87细胞分为4组,空白对照组(常规培养)和低、中、高剂量实验组(5、10、20 μmol·L-1 LCA).用细胞计数试剂盒-8检测细胞增殖能力,用克隆形成实验检测细胞克隆形成能力,用划痕实验法检测细胞迁移能力,用Transwell小室法检测细胞侵袭能力,用比色法检测总谷胱甘肱(T-GSH)、丙二醛(MDA)和超氧化物歧化酶(SOD)水平,用蛋白质印迹法检测磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)的表达水平.结果 空白对照组和低、中、高剂量实验组的细胞增殖活力分别为(90.20±2.17)％、(79.06±1.57)％、(66.13±2.11)％和(49.52±1.82)％,细胞克隆形成率分别为(76.83±2.30)％、(42.33±2.09)％、(17.71±1.84)％和(12.12±1.97)％,12 h 细胞迁移率分别为(34.92±2.24)％、(27.90±1.89)％、(18.76±1.14)％和(14.87±0.82)％,24 h 细胞迁移率分别为(50.37±2.61)％、(39.43±2.56)％、(21.11±2.33)％和(18.32±2.39)％,穿膜细胞数分别为(120.39±4.16)、(79.95±3.83)、(45.67±3.55)和(18.14±2.85)个,T-GSH 水平分别为(71.43±2.39)、(58.51±2.91)、(49.43±2.78)和(35.44±2.76)μmol·L-1,MDA 水平分别为(4.14±0.91)、(7.23±1.75)、(9.20±1.56)和(11.37±1.90)nmol·mL-1,SOD 水平分别为(41.44±2.10)、(35.43±2.91)、(28.56±2.32)和(20.62±2.05)U·mg-1,p-Akt蛋白相对表达水平分别为1.27±0.03、1.06±0.02、0.89±0.01和0.60±0.02.低、中、高剂量实验组的上述指标与空白对照组比较,在统计学上差异均有统计学意义(均P＜0.01).结论 LCA可抑制胶质瘤U87细胞的增殖、迁移、侵袭,诱导氧化损伤,其作用机制可能与下调PI3K/Akt信号通路中p-Akt蛋白的表达相关联.
Objective To investigate the effects of Licorice chalcone A(LCA)on proliferation,migration,invasion and antioxidant capacity of human glioma U87 cells and its mechanism.Methods Glioma U87 cells cultured in vitro were divided into 4 groups,blank control group(conventional culture)and experimental-L,-M,-H groups(5,10,20 μmol·L-1 LC A).Cell proliferation capacity was detected by cell counting kit-8,cell clonogenesis ability was detected by clonogenesis assay,cell migration ability was detected by scratch assay,and cell invasion ability was detected by Transwell assay.Colorimetric assay was used to detect total glutathione(T-GSH),malondialdehyde(MDA)and superoxide dismutase(SOD),and Western blotting was used to detect the protein expression levels of phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt).Results The cell proliferation activities of blank control group and experimental-L,-M,-H groups were(90.20±2.17)％,(79.06±1.57)％,(66.13±2.11)％and(49.52±1.82)％;cell clone formation rates were(76.83±2.30)％,(42.33±2.09)％,(17.71±1.84)％and(12.12±1.97)％;12 h cell mobility rates were(34.92±2.24)％,(27.90±1.89)％,(18.76±1.14)％and(14.87±0.82)％;24 h cell mobility rates were(50.37±2.61)％,(39.43±2.56)％,(21.11±2.33)％and(18.32±2.39)％;the number of perforated cells were 120.39±4.16,79.95±3.83,45.67±3.55 and 18.14±2.85;T-GSH levels were(71.43±2.39),(58.51±2.91),(49.43±2.78)and(35.44±2.76)μmol·L-1;MDA levels were(4.14±0.91),(7.23±1.75),(9.20±1.56)and(11.37±1.90)nmol·mL-1;SOD levels were(41.44±2.10),(35.43±2.91),(28.56±2.32)and(20.62±2.05)U·mg-1;the relative expression levels of p-Akt were 1.27±0.03,1.06±0.02,0.89±0.01 and 0.60±0.02,respectively.The above indexes were statistically significant between experimental-L,-M,-H groups and blank control group(all P＜0.01).Conclusion LCA can inhibit the proliferation,migration,invasion and induce oxidative damage of glioma U87 cells,and its mechanism may be related to the down-regulation of p-Akt protein expression in PI3K/Akt signaling pathway.
李红;万珊珊;刘志新;薛聪聪;李学诚;闫磊
牡丹江医学院第一临床学院临床技能中心,黑龙江牡丹江 157011黑龙江省红十字(森工总)医院消化内科,黑龙江哈尔滨 150001牡丹江医学院基础医学院,黑龙江牡丹江 157011
中医学
甘草查尔酮A;胶质瘤;迁移;侵袭;氧化损伤
Licorice chalcone A;glioma;migration;invasion;oxidative damage
《中国临床药理学杂志》 2024 (005)
678-682 / 5
黑龙江省自然科学基金资助项目(LH2020H077);牡丹江医学院科学基金火炬计划青年拔尖人才资助项目(2022-MYHJ-013);牡丹江医学院导师科研专项计划基金资助项目(YJS2X2022015)
10.13699/j.cnki.1001-6821.2024.05.010
评论