咪达唑仑对子宫平滑肌细胞钙离子移动的影响  

Effect of Midazolam on Ca^(2+) transsarcolemmal influx and Ca^(2+) release function of reticulum in human's uterine smooth muscle cells

作　　者：黄穗萍[1] 黄晓雷[1] 李元涛[1] 曹君[1] 孟馨[1] 苏娇玲[1] 胡薇[1] 

机构地区：[1]南方医科大学附属深圳妇幼保健院麻醉科,深圳518028

出　　处：《陕西医学杂志》2011年第9期1123-1126,1133,共5页Shaanxi Medical Journal

基　　金：深圳市科技计划项目(No201002083)

摘　　要：目的:探讨咪达唑仑对足月产妇子宫平滑肌细胞Ca2+跨膜内流和肌浆网Ca2+释放功能的影响及其对子宫平滑肌收缩功能影响的机制。方法:取正常孕足月待产产妇的子宫平滑肌组织,采用胶原酶消化法培养子宫平滑肌细胞,并分成两部分。第一部分:先将40个孔板中活性良好的子宫平滑肌细胞随机等分为4组(n=10),即:对照组、低浓度咪达唑仑组(M1组)、中浓度咪达唑仑组(M2组)和高浓度咪达唑仑组(M3组),用Fluo-3AM钙荧光指示剂染色后,分别给予Hank液、1μmol/L咪达唑仑(终浓度)、3μmol/L咪达唑仑、15μmol/L咪达唑仑预处理20min,然后加入40mmol/L氯化钾,每个孔板在激光共聚焦显微镜下随机选定10个子宫平滑肌细胞,利用图形分析软件分析细胞内钙荧光强度,取平均值反映子宫平滑肌细胞游离Ca2+浓度([Ca2+]i)。第二部分:以20mmol/L咖啡因代替氯化钾,其余处理及分组同第一部分。结果:第一部分与基础值比较,各组加入咪达唑仑后细胞内钙荧光强度无显著性差异(P>0.05),加入氯化钾后钙荧光强度升高(P<0.01);M1组和对照组比较加入氯化钾后钙荧光强度无显著性差异(P>0.05),M2组和M3组加入氯化钾后钙荧光强度峰值低于对照组(P<0.01),且M3组低于M2组(P<0.05)。第二部分与基础值比较,各组加入咪达唑仑后细胞内钙荧光强度无显著性差异(P>0.05),加入咖啡因后钙荧光强度升高(P<0.01);加入咖啡因后各组钙荧光强度无显著性差异(P>0.05)。结论:咪达唑仑可浓度依赖性地抑制足月产妇子宫平滑肌细胞电压依赖型钙通道开放而减少Ca2+跨膜内流,而对肌浆网Ca2+释放功能无明显影响。Objective： To study the effect of midazolam on Ca^2＋mobilization induced by KCl or caffeine in uterine smooth muscle cells intracellular,to explore its impact on uterine contractions in the possible mechanism.Methods ：The first section： Healthy full-term pregnancy from maternal uterine smooth muscle, with collagenase digestion of cultured human uterine smooth muscle cells.The cells were divided into four groups,each of 10 uterine smooth muscle cells： the control group（group C）,low concentration midazolam l（final concentration of 1μmol/L） M1 group（n=10）,middle concentration midazolam（final concentration of 3μmol/L） M2 group（n=10）,high concentration midazolam（final concentration of 15umol/L） M3 group（n=10）,with Fluo-3AM calcium fluoresceent indicator staining,respectively Hank＇s liquid,midazolam（final concentration of 1μmol/L）,midazolam（final concentration of 3μmol/L）,midazolam（final concentration of 15umol/L）.Then they were respectively preconditioned with 40mmol/L KCl.Using a confocal laser scanning microscope for determination of intracellular calcium fluorescence intensity of the dynamic changes,reflecting the intracellular free Ca^2＋concentration（ i） changes.The second section： the empirical procedure was similar to the first Section except for 20mmol/L caffeine instead of 40mmol/L KCL.Results： The first Section：Thei in Group M1,M2 and M3 were not statistically significant difference compared to group C（P〈0.05）.The[Ca^2＋]i in OHCs after addition of KCl were increased（P〈0.01）,Group M1 were not statistically significant difference compared to group C（P〈0.05）,but Group M2 and M3 were lower than group C（P〈0.01）,and Group M3 were lower than group M2 （P〈0.05）.The second section： Thei in Group M1,M2 and M3were not statistically significant difference compared to group C（P〈0.05）.Thei in OHCs after addition of caffeine were increased（P〈0.01）,but group M1,M2 and M3 were not statistically significant differen

关 键 词：子宫肌层/药物作用 子宫肌层/病理生理学 咪达唑仑/治疗应用 细胞培养 

分 类 号：R322.65[医药卫生—人体解剖和组织胚胎学]