不同浓度Mg-6Zn合金浸提液对肠上皮细胞凋亡及其相关基因Caspase-3表达的影响

发布时间：2018-10-14 19:43

【摘要】：研究不同浓度Mg-6Zn合金浸提液对大鼠肠上皮细胞IEC-6的凋亡及凋亡相关基因Caspase-3表达的影响。将Mg-6Zn合金制成不同浓度(20%和40%)的浸提液体外培养大鼠肠上皮细胞IEC-6,并以含10%胎牛血清的高糖DMEM(Dulbecco′s modified Eagle′s medium)培养基作为阴性对照。采用噻唑蓝(MTT)法通过计算细胞相对增殖率检测不同浓度合金浸提液对IEC-6细胞的毒性作用;采用流式细胞仪检测不同浓度合金浸提液对IEC-6细胞凋亡的影响;采用逆转录聚合酶链反应(RT-PCR)法检测不同浓度合金浸提液对IEC-6细胞凋亡相关基因Caspase-3mRNA表达的影响。实验结果表明,20%和40%两种浓度的镁合金浸提液在体外培养IEC-6细胞第七天时细胞毒性分级分别为0级和1级,属于无毒性,符合医用材料细胞毒性分级的合格标准。40%和20%浓度镁合金浸提液中,IEC-6细胞的凋亡比例及Caspase-3活化度相对于对照组都增高,差异有统计学意义(P0.05),40%浓度组中,IEC-6细胞的凋亡比例及Caspase-3活化度相对于20%浓度组增高,差异有统计学意义(P0.05),说明Mg-6Zn合金浸提液在一定范围内处于较高浓度时易诱导IEC-6细胞的凋亡及Caspase-3的表达,且随浓度升高影响越大。
[Abstract]:The effects of different concentrations of Mg-6Zn alloy extract on the apoptosis of IEC-6 and the expression of apoptosis-related gene Caspase-3 in rat intestinal epithelial cells were studied. Rat intestinal epithelial cells (IEC-6,) were cultured in different concentrations (20% and 40%) of Mg-6Zn alloy, and high glucose DMEM (Dulbecco's modified Eagle's medium) medium containing 10% fetal bovine serum was used as negative control. The cytotoxicity of different concentrations of alloy extract to IEC-6 cells was detected by (MTT) assay, and the apoptosis of IEC-6 cells was detected by flow cytometry. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the effect of different concentrations of alloy extract on the expression of apoptosis-related gene Caspase-3mRNA in IEC-6 cells. The results showed that 20% and 40% magnesium alloy extractions were classified as 0 and 1 respectively on the 7th day after cultured in vitro for IEC-6 cells, which were nontoxic. The ratio of apoptosis and Caspase-3 activation of IEC-6 cells in 40% and 20% magnesium alloy extracts were higher than those in control group. There was significant difference (P0.05). In 40% concentration group, the percentage of apoptosis of IEC-6 cells and the degree of Caspase-3 activation were higher than those of 20% group. The difference was statistically significant (P0.05), which indicated that Mg-6Zn alloy extract at a higher concentration could induce apoptosis and Caspase-3 expression of IEC-6 cells, and the higher the concentration was, the greater the effect was.
【作者单位】：上海交通大学附属第六人民医院普外科;南京医科大学附属无锡市第二人民医院普外科;上海交通大学材料科学与工程学院;苏州奥芮济医疗科技有限公司;
【基金】：国家自然科学基金(30901422;51271117)
【分类号】：R114

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