杜仲桃叶珊瑚苷对人神经母细胞瘤细胞增殖、迁移、凋亡及自噬的作用  

Effects of Aucubin from Eucommia Ulmoides on Proliferation,Migration,Apoptosis and Autophagy of SH-SY5Y Cells

作　　者：刘国华 杨苏芳 肖勋立 王非非 张媛 黄志强 LIU Guohua;YANG Sufang;XIAO Xunli;WANG Feifei;ZHANG Yuan;HUANG Zhiqiang(Jinggangshan University Hospital,Ji'an Jiangxi 343000,China)

机构地区：[1]井冈山大学附属医院,江西吉安343000

出　　处：《药品评价》2024年第7期825-829,共5页Drug Evaluation

基　　金：江西省教育厅科学技术研究青年项目(GJJ2201660);江西省教育厅科学技术研究一般项目(GJJ2201649)。

摘　　要：目的研究杜仲桃叶珊瑚苷(AUC)对人神经母细胞瘤(SH-SY5Y)细胞增殖、迁移、凋亡和自噬的影响,并进一步探究其作用机制。方法利用SH-SY5Y细胞构建氧糖剥夺/复氧(OGD/R)模型,设立浓度梯度,研究AUC保护OGD/R后SH-SY5Y细胞的最佳浓度。实验分为对照(Con)组、OGD/R组、AUC+OGD/R组和AUC+OGD/R+自噬激活剂(RAPA)组。光学显微镜下,观察细胞形态变化;细胞增殖检测试剂盒(CCK8)测定增殖活力;划痕愈合实验检测细胞迁移率;流式细胞术测定细胞凋亡率和Western Blot检测Beclin 1蛋白表达。结果(1)AUC的浓度确定:经低、中、高三个浓度(1μM、10μM、100μM)AUC处理OGD/R后的SH-SY5Y细胞,存活率明显升高,10μM时细胞活性最高(P<0.05),遂选取10μM的AUC进行后续实验。(2)细胞迁移能力:经AUC含药血清干预后,与OGD/R组相比,划痕的愈合率显著增加(P<0.01)。(3)细胞凋亡率:与Con组相比,OGD/R组的细胞凋亡水平显著增高(P<0.01);与OGD/R组相比,AUC+OGD/R组细胞凋亡水平显著降低(P<0.01);与AUC+OGD/R组相比,AUC+OGD/R+RAPA组细胞凋亡水平显著升高(P<0.05)。(4)Beclin 1蛋白表达:与Con组相比,OGD/R组的Beclin 1蛋白水平显著增高(P<0.01);与OGD/R组相比,AUC+OGD/R组的Beclin 1蛋白水平显著降低(P<0.01);与AUC+OGD/R组相比,AUC+OGD/R+RAPA组的Beclin 1蛋白水平显著增高(P<0.05)。结论AUC能够提高OGD/R后SH-SY5Y细胞的增殖活力和迁移愈合能力,降低凋亡率,并且对凋亡的作用可能是通过抑制自噬完成的。Objective To investigate the effects of aucubin(AUC)from eucommia ulmoides on proliferation,migration,apoptosis and autophagy of human neuroblastoma cells(SH-SY5Y),and to further explore its mechanism.Methods OGD/R model was established using SH-SY5Y cells,and the concentration gradient was set up to study the optimal concentration of SH-SY5Y cells protected by AUC(after OGD/R).The experiment was divided into control group,OGD/R group,AUC+OGD/R group and AUC+OGD/R+autophagy activator(RAPA)group.The morphological changes of cells were observed under optical microscope.Cell proliferation assay kit(CCK8)was used to determine proliferation activity.Scratch healing experiment to detect cell migration rate.Flow cytometry was used to measure cell apoptosis rate and Western Blot was used to detect Beclin 1 protein expression.Results(1)AUC concentration was determined:the survival rate of SH-SY5Y cells treated with OGD/R at low,medium and high concentrations(1μM,10μM,100μM)was significantly increased and the cell activity was the highest at 10μM(P<0.05).Therefore,AUC at 10μM was selected for the following-up experiments.(2)Cell migration ability:after the intervention of AUC drug-containing serum,the healing rate of scratches was significantly increased vs OGD/R group(P<0.01).(3)Apoptosis rate:apoptosis level in OGD/R group was significantly increased vs Con group(P<0.01);apoptosis level of AUC+OGD/R group was significantly decreased vs OGD/R group(P<0.01).Apoptosis level in AUC+OGD/R+RAPA group was significantly increased vs AUC+OGD/R group(P<0.05).(4)Beclin 1 protein expression:Beclin 1 protein level in OGD/R group was significantly increased compared with Con group(P<0.01);Beclin 1 protein level in AUC+OGD/R group was significantly decreased compared with OGD/R group(P<0.01);Beclin 1 protein level in AUC+OGD/R+RAPA group was higher than that in AUC+OGD/R group(P<0.05).Conclusion AUC can improve the proliferative activity,migration and healing ability of SHSY5Y cells after OGD/R,and reduce the apoptosis rate,and th

关 键 词：桃叶珊瑚苷 人神经母细胞瘤细胞 增殖 迁移 凋亡 自噬 

分 类 号：R285.5[医药卫生—中药学]