丙泊酚对人子宫平滑肌细胞Ca2+跨膜内流和肌浆网钙释放功能的影响

目的观察丙泊酚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,探讨其抑制子宫平滑肌收缩功能的可能机制。方法取正常孕足月待产的子宫平滑肌组织,胶原酶消化法培养子宫平滑肌细胞。①40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):对照组(C组)、低浓度丙泊酚组(P1组)、中浓度丙泊酚组(P2组)、高浓度丙泊酚组(P3组),用Fluo-3AM钙荧光指示剂染色后,分别给予Hank液、10μmol/L丙泊酚(终浓度)、50μmol/L丙泊酚、250μmol/L丙泊酚处理20 min,然后加入40 mmol/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,取钙荧光强度峰值的平均值反映子宫平滑肌细胞游离Ca2+浓度([Ca2+]i)。②再以20 mmol/L咖啡因代替氯化钾,其余处理及分组同前。结果①与基础值比较,各组加入丙泊酚后子宫平滑肌细胞[Ca2+]i差异无统计学意义(P>0.05),加入氯化钾后[Ca2+]i均升高(P<0.01);与C组相比,加入氯化钾后P1组子宫平滑肌细胞[Ca2+]i差异无统计学意义(P>0.05),P2组和P3组加入氯化钾后[Ca2+]i明显低于C组(均P<0.01),且P3组[Ca2+]i低于P2组(P<0.05)。②与基础值比较,各组加入丙泊酚后子宫平滑肌细胞[Ca2+]i差异无统计学意义(均P>0.05),加入咖啡因后[Ca2+]i升高(P<0.01);加入咖啡因后各组子宫平滑肌细胞[Ca2+]i差异无统计学意义(P>0.05)。结论丙泊酚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Ryanodine型Ca2+释放功能无明显影响。

Effect of Propofol on Ca2+ Transsarcolemmal Influx and Ca2+ Release Function of Sarcoplasmic Reticulum in Human Uterine Smooth Muscle Cells

Objective To observe the effect of propofol on Ca2+ transsarcolemmal influx and Ca2+ release function of sarcoplasmic reticulum in full-term pregnant women’s maternal uterine smooth muscle cells and to investigate its possible mechanism inhibiting uterine contraction.Methods The uterine smooth muscle tissues were obtained from healthy full-term pregnant women and human uterine smooth muscle cells were cultured by collagenase digestion methods.①The cells of 40 shadow masks were equally divided into four groups(n=10):the control group(group C),low concentration propofol group(group P1),middle concentration propofol group(group P2),high concentration propofol group(group P3),which were preconditioned with Hank liquid,10 μmol/L,50 μmol/L and 250 μmol/L propofol respectively after staining with Fluo-3AM calcium fluorescent indicator.The effects of propofol(10,50,250 μmol/L)on the changes of intracellular Ca2+ fluorescent intensity induced by 40 mmol/L KCl were investigated by a laser scanning confocal microscope.Ten uterine smooth muscle cells were randomly selected in each plate under a laser confocal microscope and the intracellular free Ca2+ concentration(i)changes were evaluated by detecting the intracellular calcium fluorescence intensity of the dynamic changes.②The empirical procedure was similar to above-mentioned except for 20 mmol/L caffeine instead of 40 mmol/L KCl.Results ①There was no significant difference in i between groups P1,P2or P3 and group C(P>0.05).The i in uterine smooth muscle cells after addition of KCl was increased(P<0.01).In the presence of KCl,there was no significant difference in i between group P1 and group C(P>0.05),but i in groups P2 and P3 was lower than in group C(P<0.01),and that in group P3 was lower than in group P2(P<0.05);②The i in groups P1,P2 and P3showed no statistically significant difference from group C(all P>0.05).In the presence of caffeine,the i in uterine smooth muscle cells was increased(P<0.01),but there was no statistically significant difference among all groups(P>0.05).Conclusion Propofol can decreases Ca2+ transsarcolemmal influx through concentration-dependently inhibiting full-term pregnant women’s uterine smooth muscle cells voltage-dependent calcium channel,but has no significant effect on Ryanodine-sensitive Ca2+ stores of endoplasmic reticulum.