四氯化碳对体外培养肝星状细胞钙浓度的影响

Effect of carbon tetrachloride on calcium concentration of hepatic stellate cells in vitro

目的研究四氯化碳（CCl4）对肝星状细胞内游离Ca^2＋（[Ca^2＋]i）的影响。方法用常规的细胞培养方法，每组加入5～15mmol／L的CCl4处理细胞，分别作用2—8h后，以Fluo-3／AM进行荧光染料负载，用激光共聚焦显微镜观察不同剂量CCl4作用不同时间[Ca^2＋]i的变化。结果CCl4在5—15mmol／L范围内可明显增加肝星状细胞[Ca^2＋]i，且[Ca^2＋]i随CCl4剂量的增大而升高（P〈0．05），作用2h后，各组[Ca^2＋]i均上升，与对照组比较差异有显著性（P〈0．05），4h后各组[Ca^2＋]i继续升高；8h后各组[Ca^2＋]i维持在较高水平，各组分别与作用4h比较差异均无显著性（P〈0．05）。结论CCl4可能引起肝星状细胞[Ca^2＋]i升高。

Objective To study the effects of carbon tetrachloride （ CCl4 ） on intracellular calcium concentration （ [ Ca^2＋ ] i） of hepatic stellate cells（HSC）. Methods Hepatic stellate cells were cultured and treated at the doses of 5 - 15 mmol/L CCl4 for 2 - 8 hours respectively. After the cells were loaded with fluorescent indicator Fluo-3/AM, [ Ca^2＋ ] i levels in HSC were measured by laser scanning confocal microscopy （LSCM）. Results At the doses of 5 - 15 mmol/L CCl4, [ Ca^2＋ ] i levels in HSC were sharply increased （ P 〈 0.05 ） and related to the dosage of CCl4. [ Ca^2＋ ] i levels in each group were increased （ P 〈 0.05 ） when HSCs were treated with CCl4 for 2 hours and had significant differences with those of the control （ P 〈 0.05 ）. [ Ca^2＋ ] i levels in HSC at 4 hours were continuously increased and remained at high stable levels at 8 hours. In comparison with [ Ca^2＋ ] i levels in HSC at the time of 4 hours, no significant difference was observed in other dose groups. Conclusion CCl4 could cause the increase of [ Ca^2＋ ] i level of hepatic steUate ceils.