壁虎粗多肽对SH-SY5Y细胞增殖和凋亡的影响

目的 研究壁虎粗多肽(GCPs)对人神经母细胞瘤(SH-SY5Y)细胞增殖、凋亡能力的影响.方法 用不同浓度GCPs(0,0.08,0.12,0.16,0.20,0.24,0.32 mg·mL-1)分别处理SH-SY5Y细胞24,48,72 h,用噻唑蓝(MTT)比色法检测SH-SY5Y细胞增殖能力的变化,根据MTT结...

Effects of Gecko crude peptides on the proliferation and apoptosis of SH-SY5Y cells

Objective To explore the effect of Gecko crude peptides(GCPs)on the proliferation and apoptosis of neuroblastoma(SH-SY5 Y)cells.Methods SH-SY5 Y cells were treated with seven different concentrations of GCPs(0,0.08,0.12,0.16,0.20,0.24,0.32 mg·mL^-1)for 24 h,48 h,72 h,respectively.Methyl thiazolyl tetrazolium(MTT)assay was used to screen out the cell proliferation.According to MTT results,the cells were divided into four groups:blank,experimental-L,-M,-H groups(GCPs:0,0.12,0.16,0.20 mg·mL^-1).Flow cytometry was used to detect the apoptosis rate of SH-SY5 Y cells.Hoechst 33258 fluorescence staining was used to detect nuclear changes.The expressions of Caspase-3 and Caspase-9 were detected by Western Blot assay.Results After treatment of GCPs for 24,48 and 72 h,the 50%inhibitory dose(IC₅₀)values were(0.23±0.01),(0.20±0.01)and(0.17±0.01)mg·m L-1,respectively.The apoptosis rates in blank,experimental-L,-M,-H groups were(10.21±0.73)%,(20.93±2.07)%,(32.90±1.07)%and(57.71±8.51)%;the apoptosis rates of experimental-L,experimental-M,and experimental-H groups were significantly higher than that of blank group(all P<0.05).Fluorescence microscopy showed that the distribution of blue fluorescence in the experimental-L,-M,-H groups were uneven,the nuclei were smaller,and there were stronger fluorophores.The protein levels of Caspase-3 in blank,experimental-L,-M,-H groups were 0.25±0.05,0.43±0.06,0.52±0.07 and0.59±0.06,respectively.Comparing between the experimental-L,-M,-H groups with the blank group,the differences were statistical(all P<0.01).The protein levels of Caspase-9 in blank,experimental-L,-M,-H groups were 0.17±0.01,0.19±0.02,0.23±0.02 and 0.26±0.02,respectively.The difference between the experimental-H group and the blank group was statistically significant(P<0.05).Conclusion GCPs can inhibit proliferation and migration of SH-SY5 Y cells,which may be associated with the Caspase pathway.