Mg-Zn-Nd-Zr镁合金浸提液对神经细胞活性和分化的影响

Effect of Mg-Zn-Nd-Zr magnesium alloy on the activity and differentiation of neural cells

目的研究Mg-Zn-Nd-Zr镁合金浸提液对神经细胞活性和分化的影响。方法制备Mg-Zn-Nd-Zr镁合金浸提液,实验分组:对照组、稀释1倍组、稀释2倍组、稀释3倍组、稀释10倍组。通过MTT法、流式细胞术、WB、CCK-8法、RT-PCR、免疫荧光法检测Mg-Zn-Nd-Zr镁合金对神经细胞活性和分化的影响。结果与对照组相比,各实验组的星型胶质细胞和神经干细胞细胞死亡率比较,差异无统计学意义(P>0.05)。各组星形胶质细胞处于G0/G1期及G2/M期的细胞比例,差异无统计学意义(P>0.05),稀释10倍组中星型胶质细胞处于S期的细胞比例减少(P<0.05)。与对照组相比,稀释10倍组和稀释3倍组中星型胶质细胞的PCNA蛋白表达量增加,而稀释2倍组和稀释1倍组中星型胶质细胞的PCNA蛋白表达量减少(P<0.05)。与对照组相比,稀释10倍组和稀释3倍组中神经干细胞的PCNAmRNA表达增加以及Bax、bcl-2的mRNA表达减少,而稀释2倍组中神经干细胞的PCNAmRNA表达减少,稀释1倍组中神经干细胞Bax、bcl-2的mRNA表达增加(P<0.05)。神经干细胞在稀释10倍组的浸提液中可分化为神经元及星形胶质细胞。结论Mg-Zn-Nd-Zr镁合金浸提液对神经细胞无明显细胞毒性。在稀释10和3倍时能促进星形胶质细胞和神经干细胞的增殖,而且稀释10倍时能促进神经干细胞向神经元及星形胶质细胞分化。但稀释2倍组和稀释1倍组的星形胶质细胞和神经干细胞凋亡增加。

Objective To investigate the effect of Mg-Zn-Nd-Zr magnesium alloy on the activity and differentiation of neural cells.Methods Preparation of different concentrations of the Mg-Zn-Nd-Zr magnesium alloy extracts,experimental groups:control group,diluted 1 times,2 times,3 times,10 times groups.The effects of Mg-Zn-Nd-Zr magnesium alloy ex⁃tracts on the activity of astrocytes were evaluated by MTT method.The cell cycle of astrocytes was detected by flow cytometry,and the expression of related proliferative proteins was detected by Western blot.The neural stem cells were tested for the cytotoxicity of the Mg-Zn-Nd-Zr magnesium alloy extracts after one day and three days by cck-8 method.The RT-PCR as⁃say extracts of each group on the gene expression of PCNA,BAX,BCL-2 of neural stem cells,neural stem cells were final⁃ly induced to differentiate at the concentrationof the Mg-Zn-Nd-Zr magnesium alloy extracts that promoted neural stem cell differentiation,and differentiated cells were identified by immunofluorescence.Results There was no statistical difference in astrocyte and neural stem cell mortality compared to controls,with the proportion of astrocytes in the 10x group increased and the decrease in phase S.PCNA protein expression was significantly increased in the 10x and 3x groups and decreased in the 2x and 1x groups.Compared with control groups,PCNA mRNA expression in Bax、bcl-2 in the 10x and 3x groups,the 2x group,and a significant increase in mRNA expression of neural stem cell Bax、bcl-2 in the 1x group.Neural stem cells can differentiate into neurons and astrocytes in 10x group extracts.Conclusions Mg-Zn-Nd-Zr magnesium alloy can pro⁃mote the proliferation of astrocytes at the dilution of 10x and PCNA protein expression at the dilution of 3x group.At the same time,Bax and bcl-2 genes were down-regulated at the dilution of 10x group.However,at the dilution of 1x and 2x group,astrocyte cell cycle and neural stem proliferation were affected by high concentrations of the Mg-Zn-Nd-Zr magne⁃sium alloy extracts.The differentiation of neural stem cells into astrocytes and neurons coμld be promoted by the dilution of 10x group of Mg-Zn-Nd-Zr magnesium alloy extract.However,apoptosis of astrocytes and neural stem cells occurred in 1x and 2x group.According to the experimental results,it can be seen that the Mg-Zn-Nd-Zr magnesium alloy has good bio⁃compatibility with nerve cells and can be used for the next animal experiment.