CAJ | 학술논문

目的：探讨人血管内皮细胞上TMEM16A、TMEM16B蛋白的表达情况，并初步探讨后者的功能。方法采用实时PCR方法检测人脐带静脉内皮细胞（human umbilical vein endothelial cells，HUVECs）中TMEM16A、TMEM16B的基因表达情况。采用Western-Blot及免疫荧光染色方法进一步证实HUVECs中TMEM16B的表达情况。Fura-2am标记HUVECs后采用Flex-station检测细胞内钙变化情况。结果 HUVECs细胞表达TMEM16B，而未表达TMEM16A。Western-Blot及免疫荧光方法从蛋白水平证明HUVECs表达TMEM16B。在使用TMEM16B的siRNA干预HUVECs后，无论是有钙及无钙溶液，在50 nM血管内皮细胞生长因子（vascular endothelial growth factor，VEGF）刺激下，钙浓度峰值均增大，而且峰值出现时间提前。结论人血管内皮细胞特异性表达TMEM16B，生理情况下TMEM16B对VEGF刺激引起的Ca2+的释放过程可能起着负向调节作用。
목적：탐토인혈관내피세포상TMEM16A、TMEM16B단백적표체정황，병초보탐토후자적공능。방법채용실시PCR방법검측인제대정맥내피세포（human umbilical vein endothelial cells，HUVECs）중TMEM16A、TMEM16B적기인표체정황。채용Western-Blot급면역형광염색방법진일보증실HUVECs중TMEM16B적표체정황。Fura-2am표기HUVECs후채용Flex-station검측세포내개변화정황。결과 HUVECs세포표체TMEM16B，이미표체TMEM16A。Western-Blot급면역형광방법종단백수평증명HUVECs표체TMEM16B。재사용TMEM16B적siRNA간예HUVECs후，무론시유개급무개용액，재50 nM혈관내피세포생장인자（vascular endothelial growth factor，VEGF）자격하，개농도봉치균증대，이차봉치출현시간제전。결론인혈관내피세포특이성표체TMEM16B，생리정황하TMEM16B대VEGF자격인기적Ca2+적석방과정가능기착부향조절작용。
ObjectiveTo investigate the expression of TMEM16A and TMEM16B protein in human vascular endothelial cells and try to clarify the function of TMEM16B.MethodsReal time PCR was used to check the expression of TMEM16A and TMEM16B in human umbilical vein endothelial cells (HUVECs). Western-Blot and immunofluorescence methods were used to confirm the expression of TMEM16B in HUVECs. Flex-station was performed to detect the change of intracellular calcium in HUVECs marked with fura-2am.ResultsHUVECs expressed TMEM16B but not TMEM16A. Western-Blot and immunofluorescence methods both proved that HUVECs expressed TMEM16B. After being intervened by siRNA of TMEM16B, with the stimulation of 50 nM VEGF, HUVECs in solution with normal Ca2+ showed not only an increased peak value but also an advanced latency of peak value. Above effect remained when HUVECs were in solution without Ca2+.ConclusionHUVECs expressed TMEM16B specifically,and it could play an negative feedback function of intracellular calcium releasing evoked by VEGF in physiological condition.