反义P38cDNA抑制胆红素诱导的人神经母细胞瘤SH-SY5Y细胞凋亡  

Antisense P38 cDNA inhibited bilir ubin-induced apoptosis in human ne uroblastoma cell line SH-SY5Y

作　　者：韩张[1] 胡萍[2] 倪道凤[1] 

机构地区：[1]中国协和医科大学中国医学科学院北京协和医院 [2]中国协和医科大学中国医学科学院基础医学研究所北京100730

出　　处：《中国民康医学》2003年第10期577-580,587,共5页Medical Journal of Chinese People’s Health

基　　金：高等学校博士学科点专项科研基金资助(No 1 9990 0 2 31 1 )

摘　　要：目的 :观察P3 8激酶特异性抑制剂SB2 0 3 580及反义P3 8激酶cDNA对胆红素诱导人神经母细胞瘤SH -SY 5Y细胞凋亡的影响 ,探讨P3 8激酶信号转导通路在胆红素诱导SH -SY5Y细胞凋亡中的作用及高胆红素血症损伤中枢神经细胞的分子机制及在听神经病发病中的作用。方法 :分别经SB2 0 3 580和二甲基亚砜预处理的SH -SY5Y细胞在胆红素作用 3h和 5h后 ,在倒置光显微镜下观察细胞形态的变化及存活情况 ;流式细胞仪检测凋亡细胞百分率。采用脂质体法将含人反义P3 8cDNA的真核表达载体 pcDNA 3 0 -antiP3 8导入SH -SY5Y细胞 ,建立稳定低表达P3 8蛋白的细胞株 ,经Westernblotting鉴定后命名为SH -SY5Y -antiP3 8,而转染空载体pcD NA3 0的SH -SY5Y细胞株则命名为SH -SY5Y -pcDNA3 0 ;SH -SY5Y -antiP3 8细胞和SH -SY5Y -pcD NA3 0细胞分别经胆红素作用 3h和 5h ,在倒置光显微镜下观察细胞形态的变化及存活情况 ;流式细胞仪检测凋亡细胞百分率。结果 :在 0 0 2 g/L胆红素作用下 ,流式细胞仪显示与对照相比SB2 0 3 580预处理的SH -SY5Y细胞在胆红素作用 3h后 ,其凋亡细胞由 19 4%下降到 12 1% ,5h后由 66 2 %降到 3 9 3 % ;与SH -SY5Y -pcDNA3 0细胞相比 ,SH -SY5Y -antiP3 8细胞在胆红素作用 3h后 。Objective:To study the role of P38 singnaling pathway in the process of bilirubin-induced a poptosis in SH-SY5Y cells Methods: Respectively pretreated with DMSO and SB203580,SH-SY5Y cells were ex posed to bilirubin,then flow cytom etry measured the number of apopto tic cells;Utilizing the antisense technique we set up cell lines wit h low expression of P38 protein wh ich confirmed by Western blotting analysis,and name it SH-SY5Y-anti P38,naming the cell line transfect ed with vector SH-SY5Y-pcDNA3 0 Th e SH-SY5Y-antiP38 cells and SH-SY5 Y-pcDNA3 0 cells were respectively exposed to bilirubin,percentage of apoptotic cells were measured by f low cytometry Results:Compare d with control the percentage of apopto tic cells in SB203580-pretreated S H-SY5Y cells decreased from 19 4% to 12 1% after treatment with bili ruin for 3h,and 66 2% to 39 3% for 5h;Compared with SH-SY5Y-pcDNA3.0 cell s the percentage of apoptotic cell s in SH-SY5Y-antiP38 cells decreased from 11 1% to 8 02% after exposed to bilirubin for 3h,and 67% to 23.4% for 5h Conclusion:P38 mediate the signal transduction of bilirubin-i nduced apoptosis in SH-SY5Y cells

关 键 词：反义P38cDNA 胆红素 诱导 神经母细胞瘤 SH-SY5Y 细胞凋亡 

分 类 号：R730.264[医药卫生—肿瘤]