摘要
目的:探索黄芪甲苷(astragaloside-Ⅳ,AS-Ⅳ)对小鼠成纤维细胞L929的毒性及影响机制,为AS-Ⅳ在皮肤创伤治疗中的应用提供实验依据。方法:体外培养L929细胞,在倒置显微镜下观察各组细胞形态,采用CCK8检测细胞增殖,Annexin V-FITC/PI双染法检测细胞凋亡,Western blotting检测细胞内cleaved Caspase-3和γ-H2AX蛋白表达,细胞划痕实验检测细胞迁移能力。结果:与Control组相比,30、60、120μmol/L的AS-Ⅳ处理L929细胞12 h、24 h,随给药浓度及给药时间增加,圆形细胞逐渐增多,悬浮细胞增多,死亡细胞逐渐增多。30、60、90、120μmol/L AS-Ⅳ处理L929细胞12 h和24 h,其OD值均低于Control组(P<0.05~P<0.01);在同一处理时间,随着给药浓度增加,其OD值逐渐降低(P<0.05~P<0.01);同一浓度AS-IV处理L929细胞12 h和24 h,其OD值均低于处理6 h(P<0.05~P<0.01)。30、60、120μmol/L AS-Ⅳ干预L929细胞24 h,细胞凋亡率均高于Control组(P<0.05~P<0.01)。与Control组相比,60μmol/L AS-Ⅳ干预L929细胞6 h、12 h、24 h均能升高细胞内cleaved Caspase-3及γ-H2AX蛋白表达(P<0.05~P<0.01);与Control组相比,30、60、120μmol/L AS-Ⅳ处理L929细胞12 h均能升高细胞内γ-H2AX和cleaved Caspase-3蛋白表达(P<0.05~P<0.01)。与Control组相比,30、60、120μmol/L AS-Ⅳ干预L929细胞12 h及24 h,相对迁移率均下降(P<0.05~P<0.01)。结论:AS-Ⅳ浓度大于30μmol/L时能抑制L929细胞增殖、迁移,诱导其凋亡。
Objective:To investigate the cytotoxicity and mechanism of astragaloside-Ⅳ(AS-Ⅳ)on L929 cells,and provide the experimental basis for the application of AS-Ⅳin treating skin wounds.Methods:The L929 cells were cultured in vitro,and the cell morphology of each group was observed under inverted microscope.The cell proliferation was detected using CCK8,and the apoptosis was detected using Annexin V-FITC/PI double staining.The expression levels of cleaved Caspase-3 andγ-H2AX protein were detected using Western blotting,and the cell migration ability was detected using cell scratch test.Results:Compared with the control group,the L929 cells were treated with 30,60 and 120μmol/L AS-Ⅳfor 12 h and 24 h,with the increasing of drug concentration and time,the number of round cells,suspended cells and dead cells increased gradually.The OD value of L929 cells treated with 30,60,90 and 120μmol/L AS-Ⅳfor 12 h and 24 h were lower than that of control group(P<0.05 to P<0.01).At the same treatment time,with the increasing of drug concentration,the OD value decreased gradually(P<0.05 to P<0.01).The OD value of L929 cells treated with the same concentration of AS-Ⅳfor 12 h and 24 h were lower than that of L929 cells treated for 6 h(P<0.05 to P<0.01).The apoptosis rate of L929 cells treated with 30,60 and 120μmol/L AS-Ⅳfor 24 h were higher than that of control group(P<0.05 to P<0.01).Compared with the control group,the expression levels of cleaved Caspase-3 andγ-H2AX protein in L929 cells treated with 60μmol/L AS-Ⅳfor 6 h,12 h and 24 h increased(P<0.05 to P<0.01).Compared with the control group,the expression levels of cleaved caspase-3 andγ-H2AX protein in L929 cells treated with 30,60 and 120μmol/L AS-Ⅳfor 12 h increased(P<0.05 to P<0.01).Compared with the control group,the relative mobility of L929 cells treated with 30,60 and 120μmol/L AS-Ⅳfor 12 h and 24 h decreased(P<0.05 to P<0.01).Conclusions:When the concentration of AS-Ⅳis higher than 30μmol/L,it can inhibit the proliferation and migration,and induce the apoptosis of L929 cells.
作者
曹静
罗时成
谈笑
程月
CAO Jing;LUO Shi-cheng;TAN Xiao;CHNEG Yue(Department of Dermatology,Lianyungang East Hospital Affiliated to Bengbu Medical College,Lianyungang Jiangsu 222042;Department of Urology,Lianyungang East Hospital Affiliated to Bengbu Medical College,Lianyungang Jiangsu 222042;Kanda College of Nanjing Medical University,Lianyungang Jiangsu 222000,China)
出处
《蚌埠医学院学报》
CAS
2021年第11期1500-1506,共7页
Journal of Bengbu Medical College
