细胞外钙对HepG2细胞内钙浓度的影响  

The effect of different concentrations of extracellular Ca^(2+) on intracellular Ca^(2+) in HepG2 cells

作　　者：王业美[1] 李松涛[1] 李颖[1] 

机构地区：[1]哈尔滨医科大学公共卫生学院营养与食品卫生学教研室,黑龙江哈尔滨150086

出　　处：《现代预防医学》2013年第12期2302-2304,共3页Modern Preventive Medicine

基　　金："863"重点项目(2010AA023002);国家自然科学基金(81102111)

摘　　要：目的研究不同浓度细胞外钙对HepG2细胞内钙的影响并初步阐明细胞内钙变化的机制。方法给予不同浓度钙培养液作用24h后,用Fluo-3/AM染色剂对HepG2细胞进行染色,应用流式细胞仪检测其细胞内钙浓度的变化;利用阻断剂对钙通道进行阻断,寻找细胞内钙变化的原因;应用siRNA干扰对找寻到的钙通道进行沉默,进一步确认引起钙变化的钙通道。结果细胞内钙浓度随细胞外钙浓度的升高而升高,当细胞外钙浓度为2.75mmol/L时,细胞内钙钙荧光强度值达到最大为(123.25±12.95);钙池操纵的钙通道(SOC)阻断剂—La3+可显著抑制外钙引起的内钙升高(P﹤0.01);应用siRNA沉默TRPC1通道后可显著抑制外钙引起的内钙变化。结论随细胞外钙浓度升高,肝细胞内钙浓度显著增高;TRPC1参与的钙池操纵的钙通道可能是引起细胞内钙离子升高的主要通道。OBJECTIVE To investigate the effect of different concentrations of extracellular Ca2＋ on intracellular Ca2. and to clarify the mechanism for the changes of intracellular Ca2＋ of HepG2 cells. METHODS HepG2 cells were cultured in different concentrations of extracellular Ca2＋ for 24 hours. After staining with Fluo-3/AM, changes of intracellular Ca2＋ were detected by flow cytometry. Ca2＋ channel blockers were used to find reasons for the changes in intracellular Ca2＋ and siRNA were used to TRPCI Ca2＋ channels. RESULTS Intracellular Ca2＋ increased according to the increase of extraeellular Ca2＋ in a dose-de- pendent manner. While the extracellular Ca2＋ was 2.75 mmol/L, intraeellular Ca2＋ reached to the maximum level of 123.25±12.95. La3＋, a store-operated Ca2＋ channels （SOCs） blocker, obviously inhibited extracellular Ca2＋ influx in HepG2 cells （P 〈 0.01 ）. When the TRPC1 was by siRNA, the increase of intracellular Ca2＋ was inhibited obviously. CONCLUSIONS Higher concentrations of extracellular Ca2＋ obviously increased intracellular Ca2＋ in HepG2 cells. TRPC1 comprised SOCs was involved in the increase of intracellular Ca2＋.

关 键 词：钙浓度 HEPG2细胞 钙通道 流式细胞仪 

分 类 号：R15[医药卫生—营养与食品卫生学]