Mg-6Zn合金对大鼠肠上皮细胞紧密连接蛋白表达的影响

Effects of Mg-6Zn alloy on tight junction protein expression in intestinal epithelial cells

目的:研究不同浓度Mg-6Zn合金浸提液对大鼠肠上皮细胞(IEC-6细胞)紧密连接蛋白表达的影响。方法:将Mg-6Zn合金制成20%和40%两个浓度的浸提液,以含10%胎牛血清的高糖DMEM培养基作为阴性对照体外培养IEC-6细胞。通过噻唑蓝(MTT)法计算细胞相对增殖率评价不同浓度Mg-6Zn合金浸提液对IEC-6细胞的毒性作用;采用逆转录聚合酶链反应(RT-PCR)法检测不同浓度Mg-6Zn合金浸提液对IEC-6细胞紧密连接相关基因ZO-1、Occludin、Claudins mRNA表达的影响。结果:20%和40%浓度Mg-6Zn合金浸提液在体外培养IEC-6细胞第7天时,细胞毒性分级分别为0级和1级,属于无毒性。20%和40%浓度Mg-6Zn合金浸提液中,IEC-6细胞紧密连接相关基因ZO-1、Occludin、Claudins mRNA的表达相对于对照组增高,差异有统计学意义(P<0.05);40%浓度组中,IEC-6细胞紧密连接相关基因相对于20%浓度组增高,差异有统计学意义(P<0.05)。结论:Mg-6Zn合金浸提液在一定浓度范围内对IEC-6细胞无毒性,对细胞紧密连接相关基因的表达有促进作用,且随浓度升高影响越大。

Objective： To evaluate the effects of different concentration of Mg-6Zn alloy extracts on tight junction protein expression in intestinal epithelial cells（IEC）-6. Methods： IEC-6 cells were cultured in 20%, 40% concentration of Mg-6Zn alloy extracts , and the Dulbecco＇s modified Eagle＇s medium-high Glucose cell medium containing 10% fetal bovine serum was served as a control group. The cytotoxicity of Mg-6Zn alloy extracts on IEC-6 cells was studied by calculating the cell relative growth rate through the MTT method. The expression of zo-1, occludin and claudins was investigated using real-time polymerase chain reaction. Results： The results showed that the cytotoxicity of the two extracts was Grade 0 1, demonstrating non-toxic to the IEC-6 cells which cultured for 7 days. The expression of tight junction related genes were up-regulated in IEC-6 cells cultured in 20% and 40% Mg-6Zn alloy extract as compared to the control group. And as compared to the 20% group, the results were also increased in 40% alloy extract. All the differences were significant （P〈0.05）. Conclusion： Mg-6Zn alloy extract at a level of concentration could promote the expression of tight junction related genes in IEC-6 cells in vitro, and the influence was more effective as the concentration of extract increased.