小檗碱、莲心碱和甲基莲心碱对HERG通道表达的影响

Effect of berberine,liensinine and neferine on HERG channel expression

目的:利用免疫荧光化学染色法和Western blot法分别定性、定量地检测不同浓度的小檗碱、莲心碱和甲基莲心碱对稳定转染在HEK-293细胞中HERG钾通道表达的影响,以及不同浓度的小檗碱对大鼠心肌组织Ikr通道蛋白表达的影响,探讨小檗碱、莲心碱和甲基莲心碱的抗心律失常机制。方法:Western blot法检测HERG-HEK细胞上HERG通道蛋白的表达;免疫荧光化学染色法并利用共聚焦显微镜定性测定不同浓度的小檗碱、莲心碱和甲基莲心碱对HERG通道蛋白表达的影响;Western blot法定量检测不同浓度的小檗碱对大鼠心肌组织Ikr通道蛋白表达的影响以及小檗碱、莲心碱和甲基莲心碱对稳定转染在HEK-293细胞中HERG钾通道蛋白表达的影响。结果:Western blot法检测稳定转染有HERG基因的HEK293细胞能高水平的表达HERG通道蛋白。10,30μmol·L-1的小檗碱明显抑制HERG-HEK细胞中HERG通道蛋白的表达(P<0.01)。10,20 mg·kg-1的小檗碱抑制大鼠心肌组织Ikr通道蛋白的表达(P<0.05)。3,10,30μmol·L-1的莲心碱促进HERG-HEK细胞中HERG通道蛋白的表达(P<0.05)。甲基莲心碱不影响HERG-HEK细胞中HERG通道蛋白的表达。结论:稳定转染的HERG-HEK细胞能高水平的表达HERG通道蛋白;小檗碱对体外HERG-HEK细胞和大鼠心肌组织Ikr通道的蛋白表达均有抑制作用;莲心碱促进HERG-HEK细胞中HERG通道蛋白的表达;甲基莲心碱对HERG通道蛋白表达无影响。

Objective： Immunofluorescence and Western blot methods were adopted for qualitative and quantitative detections of the effect of different concentrations of berberine, liensinine and neferine on the expression of stable transfection in HERG potassium channel in HEK-293 cells, as well as the effect of different concentrations of berberine on protein expression of Ikr channel in cardiac muscular tissues, in order to investigate the anti-arrhythmic mechanism of berberine, liensinine and neferine. Method： Western blot method was used to detect protein expression of HERG channel in HERG-HEK cells, hnmunoflnorescence method as well as conibcal laser microscope were used to detect the effect of different concentrations of berberine, liensinine and neferine on protein expression of HERG channel. Western blot method was used to detect the effect of different concentrations of berberine on protein expression of Ikr channel in cardiac muscular tissues as well as the effect of berberine, liensinine and neferine on protein expression of stable transfection in HERG potassium channel in HEK-293 ceils. Result： Western blot experiment manifested that stable transfection of HEK293 cells containing HERG genes could increase protein expression of HERG channel. Berberine （ 10, 30 μmol·L^-1 ） remarkably inhibited protein expression of HERG channel in HERG-HEK cells （P 〈0. 01 ）. Berberine （ 10, 20 mg·kg^-1 ） also inhibited protein expres- sion of Ikr channel in rat ventricular tissues （P 〈 0. 05）. Liensinine （3, 10, 30 μmol·L^-1 ） increased protein expression of HERG channel in HERG-HEK cells （P 〈 0. 05 ）. Neferine showed no effect on protein expression of HERG channel in HERG-HEK cells. Conclusion ： The stably transfection of HERG-HEK ceils can increase protein expression of HERG channel. Berberine shows inhibitory effect on protein expressions of in vitro HERG-HEK cells and Ikr channel in rat ventricular tissues. Liensinine improves protein expres- sion of HERG channe in HERG-HEK cells. Neferine shows no effect on protein expression of HERG channel.