抗钠钙交换体特异位点抗体对心肌钠钙交换电流的激活作用及其与钙通道、钠泵的交叉反应  

Site specific antibody of Na/Ca exchanger stimulates Na/Ca exchanger current and has cross-reaction with Ca channel and Na pump

作　　者：白晓洁[1] 吴博威[1] 

机构地区：[1]山西医科大学生理学教研室,山西太原030001

出　　处：《中国病理生理杂志》2008年第6期1078-1083,共6页Chinese Journal of Pathophysiology

摘　　要：目的:通过制备抗钠钙交换体(NCX)特异位点(α1重复序列124HNFTAGDLGPSTIVGSAAFNMF145)抗体探讨激动NCX的新途径,同时观察该抗体对于细胞膜上其它主要的离子通道和转运蛋白是否具有交叉反应。方法:将按照上述序列合成的肽段作为抗原,对健康Wistar大鼠进行连续9周的主动免疫,用SA-ELISA法检测其血清中抗体的滴度,并对抗体滴度大于1:640的样本血清进行IgGs的提取和纯化。用膜片钳全细胞电流记录方法观测在10nmol/L、20nmol/L和40nmol/L时该抗肽抗体对大鼠心室肌细胞钠钙交换电流(INa/Ca)、L-型钙电流(ICa-L)、钠泵电流(INa/kpump)、钠电流(INa)、瞬时外向钾电流(Itko)、内向整流钾电流(Ik1)以及豚鼠延迟整流钾电流(IK)的影响。结果:该抗肽抗体对于内向和外向的INa/Ca电流成分均产生剂量依赖性地激活作用,同时对IL-Ca和INa/kpump也有一定程度的激动,对检测的其余电流无显著影响。结论:该抗NCX特异位点抗体可以激动大鼠心肌钠钙交换电流,并且与L-型钙通道和钠泵具有交叉反应。AIM： To look for a new way of NCX activation by the antibody against the specific site of NCX （etl repeat sequence124 HNFTAGDLGPSTIVGSAAFNMF145 ）, and to observe the cross -reaction of the antibody to other ion channel and transporter. METHODS： Synthesized peptide as antigen was used to immunize rat for 9 weeks. The titer of the antibody was monitored by ELISA. IgGs in the positive sera （ titer 〉 1 ： 640） was purified by affinity chromatography with a Mab Trap kit. The effects of the purified IgGs on the ion channel and transporter （INa/Ca, INa pump, ICa-L, Itko, Ik1, IK ） at concentrations of 10 nmol/L, 20 nmol/L and 40 nmol/L were observed by whole cell voltage clamp technique. RESULTS： The purified IgG enhanced INa/Ca, ICa-L and INa/k pomp in a dose - dependent manner and no significant effect on INa, Itko, Ik1 and Ik was found. CONCLUSION ： The site specific antibody of Na-Ca exchanger stimulates Na-Ca exchanger current and has cross-reaction with L-type Ca channel and Na pump.

关 键 词：钠-钙交换 抗体 钙通道 钠泵 心脏 

分 类 号：R363[医药卫生—病理学]