亚砷酸钠对大鼠肝星状细胞增殖与凋亡的影响

Effects of exposure to sodium arsenite on proliferation and apoptosis in hepatic stellate cells of rats

目的探讨亚砷酸钠对大鼠肝星状细胞(HSC-T6)增殖与凋亡的影响。方法取处于对数生长期的HSC-T6细胞,调整细胞密度为4×10~4/ml,分别暴露于含终浓度为0(对照)、5、15、25μmol/L亚砷酸钠的DMEM高糖培养基24、48、72 h。采用MTT比色法检测细胞增殖活性;采用流式细胞术检测细胞早期凋亡率。结果随染毒剂量和时间增加,5μmol/L组的细胞增殖活性高于对照组,差异有统计学意义(P<0.05),25μmol/L组的细胞增殖活性低于对照组,差异有统计学意义(P<0.05)。HSC-T6细胞形态由染毒前的椭圆形和不规则形转变为染毒后的索条形,有纤维化倾向。随着染毒时间延长,染毒剂量升高,HSC-T6细胞早期凋亡率随之增高,差异有统计学意义(P<0.05),具有剂量-效应和时间-效应关系。结论低剂量亚砷酸钠暴露促进肝星状细胞增殖,高剂量亚砷酸钠暴露抑制肝星状细胞增殖,会改变细胞形态,促进细胞凋亡,具有明显的细胞毒性。

Objective To explore the effects of sodium arsenite on the proliferation and apoptosis of rat hepatic stellate cells （HSC-T6）. Methods HSC-T6 cells in logarithmic phase with the cell density of 4×10^4/ml were used, treated with nicotine at the doses of 0 （control）, 5, 15, 25 μ mol/L sodium arsenite DMEM sugar medium for 24, 48 and 72 h. The proliferation of HSC-T6 was determined by MTT assay;The early apoptosis rate of HSC-T6 was identified by flow cytometry assay. Results With the increase of exposure dose and time, ceil proliferation activity in low dose group was higher than the control group, the difference was statistically significant （P〈0.05）, in high dose group cell proliferation activity was lower than the control group, the difference was statistically significant （P〈0.05）. HSC-T6 cell morphology variated from oval and irregular deformation to bar, which had a tendency to fibrosis. As exposure time prolonged and exposure dose increased, HSC-T6 cell early apoptosis rate increased accordingly, the difference was statistically significant （P〈0.05）, with a dose-effect and time-effect manner. Conclusion Low dose exposure to sodium arsenite promotes hepatic stellate cell proliferation, high dose exposure to sodium arsenite induces inhibition of hepatic steliate cell proliferation, will change the cell morphology, promote apoptosis, has obvious cytotoxicity.