【摘要】 目的探讨氯化乙酰胆碱对体外培养的人汗腺上皮细胞内游离钙浓度的作用。方法酶消化法培养人汗腺上皮细胞。钙荧光探针Fura 3-AM染色,在原代和第1代细胞高钙或无钙培养液中加入氯化乙酰胆碱后,激光共聚焦观察其细胞内游离钙离子[Ca²⁺]；变化情况。结果细胞生长良好,约3周左右长满瓶底。细胞外液钙浓度高时（2 mmol/L）,加入氯化乙酰胆碱后原代和第1代汗腺上皮细胞的钙通道开放,钙内流,细胞内钙浓度增加,后逐渐下降,但仍高于正常。细胞外液无钙时,氯化乙酰胆碱不能刺激细胞内游离钙增加。结论体外培养的原代和第1代汗腺上皮细胞在氯化乙酰胆碱的刺激下,钙通道开放,细胞外的钙内流引起细胞内游离钙升高,而非细胞内结合钙转变成游离钙引起钙浓度增高。更多还原

【Abstract】 Objective To isolate and culture sweat gland epithelial cells in vitro,and to study the effects of acetylcholine （ACh） on intracellular flee calcium concentration ([Ca²⁺]i) of cultured sweat gland epithelial cells.Methods Sweat glands epithelial cells were collected by enzymatic digestion.After ACh was added to the primary and first passage cells,[Ca²⁺]i was examined using confocal laser scanning microscopy （CLSM） and the Ca²⁺ sensitive dye Fura 3/AM.Results The primary and first passage epithe- lial cells grew well.After ACh was added,opening of the calcium channel and significant [Ca²⁺]i increase were observed when the primary and first passage cells were incubated with high concentration of calcium （2 mmol/L）;no significant [Ca²⁺]i increase was observed in those cultured without calcium.Conclusion Upon stimulation with ACh,calcium channels of cultured primary and first passage sweat gland epithelial cells would open,influx of extracellular Ca²⁺ occurred,which resulted in an increase of [Ca²⁺]i.Extracellular bound calcium was therefore converted into intracellular free calcium.更多还原