本文關(guān)鍵詞： Brugada綜合征 SCNA基因 GC 鈉電流　出處：《南方醫(yī)科大學(xué)學(xué)報(bào)》2017年02期 　論文類型：期刊論文

【摘要】：目的探討B(tài)rugada綜合征SCN5A基因新突變G1712C的電生理機(jī)制。方法采用體外定點(diǎn)誘變法構(gòu)建攜帶有基因突變G1712C的p Rc/CMV-h H1的表達(dá)載體,lipo3000脂質(zhì)轉(zhuǎn)染法建立穩(wěn)定表達(dá)p GFP-IRES-hβ1質(zhì)粒的HEK293細(xì)胞系,并用G418進(jìn)行篩選鑒定。分別做野生型的p Rc/CMV-h H1(h H1)和攜帶有基因突變G1712C的p Rc/CMV-h H1(mh H1)瞬時(shí)轉(zhuǎn)染表達(dá)。進(jìn)行全細(xì)胞膜片鉗實(shí)驗(yàn)記錄鈉通道電流。實(shí)驗(yàn)結(jié)果由Patch Master以及IGOR Pro 6.0軟件分析。結(jié)果 G1712C位于Na+通道蛋白α亞單位的DⅣ區(qū)S5與S6之間的P-loop上。在瞬時(shí)轉(zhuǎn)染野生型的h H1的細(xì)胞系中,指令電位從-60 m V逐漸上升時(shí),鈉電流也漸變大,在-20 m V時(shí)完全激活;激活電壓在-60 m V到-50 m V,反轉(zhuǎn)電位在50 m V左右。在瞬時(shí)轉(zhuǎn)染突變型G1712C的細(xì)胞系中,沒有發(fā)現(xiàn)鈉電流。結(jié)論野生型h H1所表達(dá)的鈉通道蛋白與正常心肌細(xì)胞鈉通道電生理特性相似。SCN5A基因G1712C突變導(dǎo)致Nav1.5通道失去功能,可能是該家系Brugada綜合征的病因。
[Abstract]:Objective to investigate the electrophysiological mechanism of G1712C, a new mutation of SCN5A gene in Brugada syndrome. Methods to construct the gene mutation G1712C by site-directed mutagenesis in vitro. The expression vector of Rc/CMV-h H1. HEK293 cell lines stably expressing p GFP-IRES-h 尾 1 plasmid were established by lipo3000 lipid transfection. Using G418 to screen and identify. The wild type p Rc/CMV-h H1 1 and the gene mutation G1712 C carrying p Rc/CMV-h 1 1 were made respectively. Mh 1). Transient transfection expression. The sodium channel currents were recorded by whole-cell patch clamp experiments. The results were analyzed by Patch Master and IGOR Pro 6.0 software. G1712C is located on the P-loop between S5 and S6 in the D 鈪，

本文編號(hào)：1482499