壁虎粗多肽对SH-SY5Y细胞增殖和凋亡的影响  

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

作　　者：尹慧萍 王兵兵 黄泽月 王艳 王建刚[1] YIN Hui-ping;WANG Bing-bing;HUANG Ze-yue;WANG Yan;WANG Jian-gang(Medical College,Henan University of Science and Technology,Luoyang 471023,Henan Province,China;Department of Pharmacy,The First Affiliated Hospital of Henan University of Science and Technology,Luoyang 471000,Henan Province,China)

机构地区：[1]河南科技大学医学院,河南洛阳471023 [2]河南科技大学第一附属医院药学部,河南洛阳471000

出　　处：《中国临床药理学杂志》2021年第6期681-684,共4页The Chinese Journal of Clinical Pharmacology

基　　金：河南省科技重点攻关基金资助项目(142102310031)。

摘　　要：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结果分组。将SH-SY5Y细胞分为空白组和低、中、高剂量实验组(GCPs:0,0.12,0.16,0.20 mg·mL^(-1))。用流式细胞仪检测SH-SY5Y细胞的凋亡率,用Hoechst 33258荧光染色检测细胞核变化,用蛋白质印迹法检测胱天蛋白酶-3(Caspase-3)和Caspase-9蛋白的表达水平。结果GCPs作用SH-SY5Y细胞24,48,72 h的半数抑制浓度(IC_(50))值分别为(0.23±0.01),(0.20±0.01)和(0.17±0.01)mg·mL^(-1)。空白组和低、中、高剂量实验组的细胞凋亡率分别为(10.21±0.73)%,(20.93±2.07)%,(32.90±1.07)%和(57.71±8.51)%,低、中、高剂量实验组的凋亡率与空白组比较,差异均有统计学意义(均P<0.05)。在荧光显微镜下观察,低、中、高剂量实验组蓝色荧光分布不均,细胞核缩小,有较强荧光团出现。空白组和低、中、高剂量实验组的Caspase-3与GAPDH灰度值比值分别为0.25±0.05,0.43±0.06,0.52±0.07和0.59±0.06,低、中、高剂量实验组与空白组比较,差异均有统计学意义(均P<0.01);Caspase-9与GAPDH灰度值比值分别为0.17±0.01,0.19±0.02,0.23±0.02和0.26±0.02,高剂量实验组与空白组比较,差异有统计学意义(P<0.05)。结论GCPs可抑制SH-SY5Y细胞增殖并诱导其凋亡,其机制可能与激活Caspase凋亡途径相关。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(IC_(50))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.

关 键 词：壁虎粗多肽 SH-SY5Y细胞 增殖 凋亡 

分 类 号：R28[医药卫生—中药学]