摘要
目的:探讨持续高浓度葡萄糖干预对Kv11.1离子通道蛋白表达的影响。方法:①采用双酶切法和基因重建技术将HERG基因插入到表达绿色荧光蛋白的真核表达载体pEGFP-N1中,构建Kv11.1离子通道蛋白的表达载体pEGFP-N1-HERG并测序验证。②pEGFP-N1-HERG表达载体鉴定成功后经脂质体转染HEK293T细胞,并通过不同糖浓度(5、17.5、30mmol/L)干预细胞48h后流式细胞仪检测细胞HERG离子通道蛋白绿色荧光平均表达量。结果:流式细胞仪检测pEGFP-N1-HERG融合蛋白平均荧光强度于不同浓度葡萄糖持续干预后分别为218.87(5mmol/L)、174.83(17.5mmol/L)、142.90(30mmol/L),三组间比较差异有统计学意义(均P<0.05)。结论:持续高糖状态抑制Kv11.1离子通道蛋白的表达,为糖尿病患者长期高糖状态时QT间期延长提供理论依据并奠定实验基础。
Objective:To investigate the effect of continuous high concentration glucose intervention on the expression of Kv11.1 ion channel protein.Methods:①The HERG gene was inserted into the eukaryotic expression vector pEGFP-N1 expressing green fluorescent protein by double enzyme digestion method and gene reconstruction technology to construct the expression vector pEGFP-N1-HERG of Kv11.1 ion channel protein and sequence it for verification.②After the identification of pEGFP-N1-HERG expression vector was successful,HEK293T cells were transfected with liposomes,and the average expression of HERG ion channel protein green fluorescence was detected by flow cytometry after 48 hours of intervention with different glucose concentrations(5,17.5,30 mmol/L).Results:The average fluorescence intensity of pEGFP-N1-HERG fusion protein detected by flow cytometry was 218.87(5 mmol/L),174.83(17.5 mmol/L),142.90(30 mmol/L)respectively after continuous intervention with different concentrations of glucose.There was significant difference among the three groups(P<0.05).Conclusion:Sustained hyperglycemia inhibits the expression of Kv11.1 ion channel protein,which provides theoretical basis and experimental basis for QT interval extension in diabetes patients with long-term hyperglycemia.
作者
韩稳琦
王毅
陈海潮
尤红俊
邓纪钊
祁杰
HAN Wenqi;WANG Yi;CHEN Haichao;YOU Hongjun;DENG Jizhao;QI Jie(Second Department of Cardiovascular Medicine,Shaanxi Provincial People’s Hospital,Xi’an 710068,China)
出处
《陕西医学杂志》
CAS
2024年第1期32-36,共5页
Shaanxi Medical Journal
基金
陕西省科学技术研究发展计划资助项目(2022SF-152)
陕西省人民医院科技人才支持计划项目(2022JY-69)
陕西省人民医院科技发展孵化基金资助项目(2022YJY-53)。
