中华皮肤科杂志

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氯化乙酰胆碱对培养的汗腺上皮细胞内游离钙的调节作用

雷霞1, 伍津津1, 鲁元刚1, 朱堂友1, 龙在云2, 刘媛2   

  1. 1. 第三军医大学大坪医院野战外科研究所皮肤科 重庆 400032;
    2. 第三军医大学大坪医院野战外科研究所三室
  • 收稿日期:2005-11-08 出版日期:2006-04-15 发布日期:2006-04-15
  • 通讯作者: 伍津津, email:wjjj@163.com E-mail:wjjj@163.com
  • 基金资助:
    国家863计划资助项目(2003AA205020);国家自然科学基金(30570972)

Effects of acetylcholine chloride on intracellular free calcium concentration of cultured sweat gland epithelial cells

LEI Xia1, WU Jin-jin1, LU Yuan-gang1, ZHU Tang-you1, Long Zai-yun2, LIU Yuan2   

  1. Department of Dermatology, Daping Hospital, Third Military Medical University, Chongqing 400042, China
  • Received:2005-11-08 Online:2006-04-15 Published:2006-04-15

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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+]i;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。

关键词: 汗腺, 钙, 乙酰胆碱

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 epithelial 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.

Key words: Sweat glands, Calcium, Acetylcholine