槟榔碱促结肠平滑肌细胞收缩及对胞内钙离子浓度的影响  

Effects of Arecolinehydrobromide on contraction and intracellular free Ca^(2+) concentrations of colonic smooth muscle cells

作　　者：梁宁霞[1] 衣兰娟[1] 田琳[1] 胡凡[2] 刘翠萍[3] 魏睦新[1] 

机构地区：[1]南京医科大学第一附属医院中医科,江苏省南京市210029 [2]南京医科大学,江苏省南京市210029 [3]南京医科大学第一附属医院内分泌科,江苏省南京市210029

出　　处：《世界华人消化杂志》2006年第7期676-681,共6页World Chinese Journal of Digestology

基　　金：江苏省科技厅国际合作项目;No.BZ2002066江苏省"135"工程重点人才项目;No.RC2002049

摘　　要：目的:观察氢溴酸槟榔碱(Ah)促进培养的结肠平滑肌细胞收缩作用及对胞内游离Ca2+浓度的影响．方法:培养大鼠结肠平滑肌细胞,分为4组:正常对照组、Ah刺激组、乙酰胆碱(Ach)刺激组、阿托品预处理组．应用特异性Ca2+荧光批示剂Fluo-3／AM负载细胞,激光共聚焦显微镜检测游离Ca2+浓度和细胞收缩率．结果:正常对照组细胞未发生自主性收缩,因指示剂的衰减,荧光强度(FI)有递减的趋势; Ah刺激组在加药后短时间内胞内钙离子浓度迅速升高,出现一个波峰,FI平均基础值与峰值有差异(P<0．05),而后胞内钙离子浓度再缓慢攀升,在484．0±47．6 s达到高峰,而后有一个较长时间的平台期,在1 400 s左右恢复至静息水平,FI基础值与峰值有显著差异 (P<0．01),细胞收缩百分数为20．70％±0．07％; Ach刺激组在加药后胞内钙离子浓度升高,形成一个小波峰,FI基础值和峰值差异有显著性(P<0．01)．随后胞内钙离子浓度缓慢攀升, 在600 s左右达到高峰,而后有一个较长时间的平台期．FI基础值与峰值差异有显著性 (P<0．001);经阿托品预孵育处理后的细胞,加入Ah后,其收缩效应被完全抑制．结论:Ah可引起结肠平滑肌细胞收缩,升高细胞内游离Ca2+浓度．其收缩效应可以被M受体阻断剂阿托品所抑制．AIM： To explore the possible role of Arecolinehydrobromide （Ah） in the dynamics of digestive tract and the effects of Ah on the contraction of colon smooth muscle cells （SMCs） and the concentration of intraceUular [Ca^2＋]i. METHODS： SMCs from colon of mice were cultured in vitro. Based on different treatment methods, four groups were divided into as follows： Ah group in which cultured SMCs were stimulated by Ah; control group in which there was no additional treatment on cultured SMCs; Acetylcholine （Ach） group in which cultured SMCs were stimulated by Ach; Atropine group in which cultured SMCs were pretreated with atropine before Ah was added. Laser scanning confocal microscope （LSCM） was used to measure the concentration of intracellular [Ca^2＋]i and the contraction of colon SMCs in the above four groups. RESULTS： No spontaneous cellular contraction took place in control group in which there was a tendency of decrease in fluorescent intensity because of attenuation of the indicator. In Ah group, a rapid elevation in the concentration of intracellular [Ca^2＋]i occurred shortly after Ah was added, which was recorded as a pulse wave. Then there was a gradual and slow increase in intracellular [Ca^2＋]i which reached its peak at 484.0 ± 47.6 s. The peak was followed by a quite long plateau and afterward returned to the resting level at 1 400 s. The percentage of cellular contraction was 20.70% ± 0.07%. In Ah and Ach groups, a tendency of SMCs contraction in accordance with elevation of intraceUular [Ca^2＋]i was observed following Ah or Ach was added. However, the effect was faster and shorter in Ah group than that in Ach group. The contractive effect of Ah was completely inhibited in SMCs pretreated by atropine. CONCLUSION： Ah can cause contraction of SMCs of colon and increase of intracellular [Ca^2＋]i. The contractive effect of Ah can be effectively inhibited by atropine. Ah may be promising for improving dynamics of digestive tract.

关 键 词：结肠 平滑肌细胞 细胞培养 槟榔碱 钙离子 激光共聚焦显微镜 

分 类 号：R285.5[医药卫生—中药学]