CD147超表达对γ分泌酶影响的临床研究

Effect of over expressed CD147 on γ-secretase

目的探讨CD147在小鼠胚胎成纤维细胞中超表达对γ分泌酶活性的影响。方法 CD147cDNA转染敲除淀粉样前体蛋白表达基因的小鼠胚胎成纤维细胞系,以梯度浓度G418筛选CD147表达量最高的细胞,并提取实验组样本E1、E2及IPE2和对照组C1、C2和IPC2中γ分泌酶复合物。蛋白质免疫印迹法(Western blotting)检测CD147的表达量,酶联免疫吸附测定(ELISA)分析样品Aβ40和Aβ42的产量。结果样本E1、E2及IPE2中CD147表达量较相应对照组样本C1、C2和IPC2分别上升(11.0±5.5)%(P=0.08),(28.2±9.1)%(P<0.01)及(50.0±9.8)%(P<0.002),γ分泌酶活性分别下降(61.8±3.0)%(P=0.25),(82.9±5.0)%(P<0.001),(190±7)%(P<0.001)。结论 CD147是γ分泌酶复合物的负调控因子。

Objective Construct the model of CD147 over-expression and research the effect of CD147′s over expression in mouse embryonic fibroblasts on γ-secretase.Methods MEF（KO） with the amyloid precursor protein gene knockout was transfected with CD147 cDNA.Use the G418 to choose the cells with the most expressed CD147,and then extract γ-secretase from E1,E2 and IPE2 in the experiment group and C1,C2 and IPC2 in the control group.The protein levels of CD147 were detected by Western blotting,the output of Aβ40 and Aβ42 were detected by ELISA.Results Compared with C1,C2 and IPC2,the protein levels of CD147 in E1,E2 and IPE2 were respectively increased by（11.0±5.5）%（P=0.08）,（28.2±9.1）%（P0.01）and（50.0±9.8）%（P0.002）.However,activity of γ-secretase was respectively declined by（61.8±3.0）%（P=0.25）,（82.9±5.0）%（P0.001） and（190±7）%（P0.001）.Conclusion CD147 is a negative regulator of the γ-secretase complex.