SHSY5Y神经细胞中尼古丁受体α3亚单位基因表达抑制与APP代谢的关系临床医学.docVIP

SHSY5Y神经细胞中尼古丁受体α3亚单位基因表达抑制与APP代谢的关系临床医学 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：SHSY5Y神经细胞中尼古丁受体α3亚单位基因表达抑制与APP代谢的关系临床医学 1 1 材料与方法 3 2 结 果 5 3 讨 论 6 文2：人参水提物对β淀粉样蛋白诱导SHSY5Y细胞凋亡的保护作用 7 1 材料与方法 8 2 结果 10 3 讨论 12 参考文摘引言： 13 原创性声明（模板） 14 文章致谢（模板） 15 正文 SHSY5Y神经细胞中尼古丁受体α3亚单位基因表达抑制与APP代谢的关系临床医学 文1：SHSY5Y神经细胞中尼古丁受体α3亚单位基因表达抑制与APP代谢的关系临床医学 【Abstract】 Objective To investigate the influence of inhibited gene expression of α3 nicotinic acetylcholine receptor (nAChR) induced by RNA interference on the metabolism of (amyloid precuor peptide (APP). Methods The siRNA coding oligonucleotide sequences targeting α3 nAChR were designed and synthesized. The annealed product was cloned into pSliencer H1 neo vector. The recombinant α3 nAChR pSilencer H1 neo vector was trafected into the SHSY5Y cells. The stable clones were screened by G418 medium, and the levels of α3 nAChR mRNA and protein were monitored by using Realtime PCR and Western blotting respectively. The levels of the (form of secreted APP ((APPs) and total APP were also determined by Western blotting. Results Compared with control, the expression levels of mRNA and protein in the SHSY5Y cells trafected with the recombinant α3 nAChR pSilencer H1 neo vector were decreased by the inhibitory efficiency with 98% and 66% respectively. The protein level of the αAPPs was reduced and no significant change of total APP was observed. Conclusio The inhibited gene expression of α3 nAChR resulted from the recombinant α3 nAChR siRNA can decrease cleavage of APP by (secretase, which suggest that α3 nAChR may play a significant neuroprotective role in the pathogenesis of Alzheimer′s disease. 【Key words】 Silence interference RNA; α3 nAChR; Neuroblastoma cell; Amyloid precuor peptide 神经型尼古丁受体（nAChR）与大脑学习记忆、智力发育和识别能力等脑功能有关，还具有明显的神经保护作用〔1～3〕。研究表明，尼古丁受体与淀粉样前体蛋白(APP) 的代谢过程有密切联系，在阿尔茨海默病(AD)的发病中起着重要的作用〔2〕。小干扰RNA(siRNA)能特异性结合并切割同源性mRNA，是转录后基因沉默现象的重要机制之一〔4〕。运用siRNA真核表达载体在哺乳动物细胞内合成siRNA能高效抑制特异性的基因表达〔5，