氯化钴对小鼠NIH3T3细胞增殖和凋亡的影响

Effects of Cobalt Chloride on Proliferation and Apoptosis of NIH3T3 Cells in Mice

目的:观察氯化钴(Cocl2)对体外培养的小鼠NIH3T3细胞增殖和凋亡的影响。方法:首先利用MTT法和流式细胞术检测不同浓度Cocl2对小鼠NIH3T3细胞增殖及凋亡的影响。然后利用免疫印迹检测HIF-1α和凋亡相关蛋白Bcl-2及Bax的表达。结果:(1)MTT结果显示,低于250μmol/L的Cocl2作用NIH3T3细胞24h对细胞的增殖活力影响不大,当浓度增加到500μmol/L以上时会明显抑制细胞的增殖活力。(2)流式细胞仪检测结果显示低浓度Cocl2(≤250μmol/L)并不会引起NIH3T3细胞明显的凋亡和坏死,增加浓度(≥500μmol/L)会导致细胞早期凋亡和坏死增加。3)不同浓度Cocl2作用NIH3T3细胞24h均可诱导细胞中HIF-1α表达上调,同时Bax/Bcl-2的比值增高。结论:高浓度Cocl2可以抑制小鼠NIH3T3细胞增殖,促进凋亡发生。

Objective：To study effects of Cocl2 on survival,proliferation and apoptosis of mouse NIH3T3 cells.Methods：The survival,proliferation and apoptosis effects of different concentrations of Cocl2 on mouse NIH3T3 were detected by MTT and flow cytometer assay.The expression of HIF-1α,Bcl-2 and Bax protein were detected by Western blot.Results：（1） MTT assay showed that the cellular proliferation activity was inhibited significantly by higher concentration（≥500 μmol/L） of Cocl2,however lower concentration（250 μmol/L） of Cocl2 had little effects on the proliferation of NIH3T3.（2） Flow cytometer assay showed higher concentration of Cocl2 increased the early apoptosis and necrosis rates of NIH3T3,but the lower concentration of Cocl2 could not affect the apoptosis and necrosis rates.（3） The expression of HIF-1α protein could be upregulated by different concentrations of Cocl2 in NIH3T3 cells and the expression ratio of Bax/Bcl-2 was increased.Conclusions：Higher concentrations of Cocl2 could inhibit the proliferation and promote the apoptosis of mouse NIH3T3 cells.