Study on the toxicity of astragaloside-Ⅳ to fibroblasts

ObjectiveTo investigate the cytotoxicity and mechanism of astragaloside-Ⅳ(AS-Ⅳ) on L929 cells, and provide the experimental basis for the application of AS-Ⅳ in treating skin wounds.

MethodsThe 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 and γ-H2AX protein were detected using Western blotting, and the cell migration ability was detected using cell scratch test.

ResultsCompared with the control group, the L929 cells were treated with 30, 60 and 120 μmol/L AS-Ⅳ for 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 120 μmol/L AS-Ⅳ for 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-Ⅳ for 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 120 μmol/L AS-Ⅳ for 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 and γ-H2AX protein in L929 cells treated with 60 μmol/L AS-Ⅳ for 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 and γ-H2AX protein in L929 cells treated with 30, 60 and 120 μmol/L AS-Ⅳ for 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 120 μmol/L AS-Ⅳ for 12 h and 24 h decreased(P < 0.05 to P < 0.01).

ConclusionsWhen the concentration of AS-Ⅳ is higher than 30 μmol/L, it can inhibit the proliferation and migration, and induce the apoptosis of L929 cells.