不同浓度钙对人腹膜间皮细胞影响的实验研究

Effect of Various Concentration Calcium on Human Peritoneal Mesothelial Cells

【目的】观察不同浓度钙对体外培养的人腹膜间皮细胞(human peritoneal mesothelial cells,HPMCs)及细胞外基质的影响。【方法】采用人腹膜间皮细胞株体外培养,用乳酸脱氢酶(LDH)和四甲基偶氮唑盐比色法(MTT)评估细胞的活力和增殖能力;采用免疫细胞化学法来检测纤维连接蛋白(FN)的表达。【结果】不同浓度钙均可促进HPMCs增殖(P<0.01),而以1.25 mmol/L及1.0 mmol/L钙作用最明显,且呈时间依赖关系。不同浓度钙组与对照组相比均可明显升高LDH水平,其中以1.25 mmol/L钙的LDH水平最低(P<0.01),且呈时间依赖关系。2.0 mmol/L和1.75 mmol/L钙可明显上调FN的表达(P<0.01)。【结论】高钙可明显损伤HPMCs及促进FN合成,其在腹膜纤维化的发生中可能起了一定作用;而浓度为1.25 mmol/L钙可保护腹膜,并在一定程度上有预防腹膜纤维化的作用。

[ObjectivelTo study the effect of calcium on human peritoneal mesothelial cells （HPMCs）, [Methods]Proliferation abilities of HPMCs were assessed by tetrazolium salt colorimetry assay （MTT assay） and the levels of LDH in the supernatant were detected in all groups to evaluate the damage of HPMCs. The expression of fibronectin（FN） in cytoplasm was detected by Immunohistochemistry. [Resuhs]Calcium enhanced proliferation of HPMCs was time-dependence（ P〈0. 01）, especially calcium with 1.25mmol/L and 1.0 mmol/L, Calcium with different concentrations significantly increased the levels of LDH in HPMCs in a tlme-dependence, while the effect of calcium with 1.25 mmol/L was lightest（ P〈0.01）. Calcium with 2.0 mmol/L and 1.75mmol/L significantly up-regulated the expression of FN（ P〈0, 01） . [Conclusion]High calcium damages HPMCs and up-regulates the expression of FN by HPMCs, while low calcium （1.25 mmol/L） can protect peritoneum and prevent its fibrosis.