Inhibitory effect and mechanism of sodium citrate on calcification of mouse vascular smooth muscle cells induced by high phosphorus

doi:10.3760/cma.j.issn.1007-1245.2021.06.019

Abstract

Abstract: Objective To investigate the effect of sodium citrate (Na₃Cit) on calcification of vascular smooth muscle cells (MOVAS) in mice with chronic kidney disease (CKD) induced by high phosphorus and its mechanism, and to search for drugs that are anticoagulant and can improve vascular calcification (VC) of CKD. Methods MOVAS was cultured in high phosphorus and different concentrations of Na₃Cit for 14 days. Calcium deposition was detected by alizarin red staining and o-cphenolphthalein complex ketone method; phenotype transformation was detected by real-time fluorescent quantitative PCR and alkaline phosphatase (ALP) staining; apoptosis was detected by annexin V staining. Results Na₃Cit significantly inhibited the calcification of CKD MOVAS induced by high phosphorus, and the inhibitory effect of high concentration of Na₃Cit was better. Na₃Cit could inhibit calcium and phosphorus deposition by forming calcium citrate chelate, thus inhibiting calcification. Na₃Cit could also inhibit ALP activity and up-regulate SM22-α expression of smooth muscle cells under high phosphorus conditions, thus preventing smooth muscle cells from differentiating into osteoblast like phenotype; meanwhile, it could reduce apoptosis induced by high phosphorus to inhibit calcification. Conclusion Na₃Cit can effectively reduce the calcification of MOVAS induced by high phosphorus. As a blood anticoagulant, Na₃Cit may play an anti VC role in clinic.

Key words: Chronic kidney disease, Vascular calcification, High phosphorus, Sodium citrate

摘要： 目的探究枸橼酸钠(Na₃Cit)对高磷诱导的慢性肾脏病(CKD)小鼠血管平滑肌细胞(MOVAS)钙化的影响及其机制,寻找既能抗凝又能改善CKD血管钙化(VC)的药物。方法在高磷及不同浓度Na₃Cit条件下培养MOVAS 14 d,通过茜素红染色和邻甲酚酞络合酮法检测细胞钙沉积量;实时荧光定量PCR和碱性磷酸酶(ALP)染色检测细胞表型转化;annexin V染色检测细胞凋亡情况。结果 Na₃Cit能明显抑制高磷诱导的CKD MOVAS钙化,高浓度Na₃Cit抑制效果更好。Na₃Cit能够通过形成枸橼酸钙螯合物抑制钙磷的沉积从而抑制钙化。Na₃Cit还能抑制高磷条件下ALP的活性及上调平滑肌细胞标示物SM22-α的表达,从而阻止平滑肌细胞向成骨样细胞表型的分化,同时能减小高磷诱导的细胞凋亡来抑制钙化。结论 Na₃Cit能有效减少高磷诱导的MOVAS钙化。作为血液抗凝剂的Na₃Cit有可能在临床中发挥抗VC的作用。

关键词: 慢性肾脏病, 血管钙化, 高磷, 枸橼酸钠

Chen Hui, Yu Hui. Inhibitory effect and mechanism of sodium citrate on calcification of mouse vascular smooth muscle cells induced by high phosphorus[J]. International Medicine and Health Guidance News, 2021, 27(6): 861-868.

陈辉, 余辉. 枸橼酸钠对高磷诱导小鼠血管平滑肌细胞钙化的抑制作用及机制[J]. 国际医药卫生导报, 2021, 27(6): 861-868.

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