摘要
目的探讨腺病毒介导的人Na+-H+交换体-1(NHE-1)miRNA转染人源性神经母细胞瘤细胞株(SH-SY5Y)后,细胞内pH值变化及其对β位淀粉样前体蛋白裂解酶1(BACE1)的影响。方法培养SH-SY5Y,随机分为正常对照组、NHE-1基因敲低组和阴性对照组,并构建NHE-1基因敲低细胞模型。采用荧光分光光度计测定各组细胞内pH值,通过Real-time PCR及Western blotting检测BACE1 mRNA及蛋白表达。结果与正常对照组相比,NHE-1基因敲低组细胞内pH值明显降低(P<0.01),阴性对照组细胞内pH值无明显变化;NHE-1基因敲低组细胞内BACE1 mRNA表达及蛋白含量明显增高(P<0.01),阴性对照组细胞内BACE1 mRNA表达及蛋白含量无明显变化。结论细胞内pH值降低能够增强BACE1的表达。
Objective To transfect the adenovirus mediated NHE-1 miRNA into human neuroblastoma（ SH-SYSY）, and investigate the expression of BACE1 with the condition of the change of intracellular pH value. Methods The human neuroblastoma （SH-SYSY） cell were cultured, enrolled and randomly divided into 3 groups, named as normal control group, NHE-1 gene knock down group and negative control group, and structured the NHE-1 gene knock down model. The intracellular pH value was detected in each group by fluorescence spectrophotometer, cells were examined for mRNA and protein expression of BACEI by Real-time PCR and Western blotting. Results Compared with control group, the intracel- lular pH value was significantly lower in NHE-1 gene knock down group by fluorescent probe method （P 〈 0.01 ）, while it had no change in negative control group; furthermore, the BACE1 mRNA expression and protein level were significantly higher in NHE-1 gene knock down group than those in paired normal control group （P 〈0.01 ）, the BACEI mRNA expres- sion and protein level had no change in negative control group. Conclusions The lower intracellular pH value condition is responsible for the higher BACEI expression.
出处
《山东医药》
CAS
2012年第9期1-3,共3页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81100962)
辽宁省博士科研启动
自然科学基金资助项目(20082179)
辽宁省教育厅高等学校科研项目(20060524)
