枸橼酸二乙酯、枸橼酸钠和膦甲酸钠对高钙诱导小鼠血管平滑肌细胞钙化的抑制  

Inhibition of Diethyl Citrate,Sodium Citrate,and Phosphonoformic Acid on Mouse Aortic Smooth Muscle Cells Calcification Induced by High Calcium

作　　者：黄芳 张冲宇 黄玲红 欧阳健明[1] HUANG Fang;ZHANG Chong-Yu;HUANG Ling-Hong;OUYANG Jian-Ming(Institute of Biomineralization and Lithiasis Research,Jinan University,Guangzhou 510632,China)

机构地区：[1]暨南大学生物矿化与结石病防治研究所,广州510632

出　　处：《无机化学学报》2020年第7期1353-1365,共13页Chinese Journal of Inorganic Chemistry

基　　金：国家自然科学基金(No.21975105)资助项目。

摘　　要：在高钙及不同浓度枸橼酸二乙酯(Et2Cit)、枸橼酸钠(Na3Cit)和膦甲酸钠(PFA)存在下培养小鼠血管平滑肌细胞(MOVAS)14 d,分别通过茜素红染色、免疫荧光和annexin V染色检测细胞的分化、凋亡和细胞钙沉积量,研究了Et2Cit、Na3Cit和PFA对高钙诱导MOVAS细胞钙化的抑制效果及可能的作用机制。结果表明,Et2Cit、Na3Cit和PFA均能减少高钙诱导的MOVAS钙化,减少细胞外钙化斑块和钙沉积量。这些抑制剂均能抑制平滑肌细胞向成骨样细胞表型转化,导致向成骨细胞转化的标记物碱性磷酸酶(ALP)活性降低。抑制效果均存在浓度依赖性。当抑制剂浓度相同时,其抑制效果从大到小为:PFA>Na3Cit>Et2Cit。低浓度的Et2Cit和Na3Cit可通过减少细胞凋亡来抑制钙化,但高浓度的Et2Cit、Na3Cit和PFA自身具有毒性,这增加了细胞的凋亡。作为血液抗凝剂的Et2Cit和Na3Cit可以有效地抑制MOVAS的钙化。Mouse aortic smooth muscle cells(MOVAS)were cultured for 14 d under the conditions of high calcium and different concentrations of diethyl citrate(Et2Cit),sodium citrate(Na3Cit),and phosphonoformic acid(PFA).Alizarin red staining,immunofluorescence and annexin V staining were used to detect cell transdifferentiation,apoptosis,and cell calcium deposition.The inhibitory effects of Et2Cit,Na3Cit,and PFA on calcium-induced calcifi-cation of MOVAS cells and their possible mechanisms were studied.The results showed that Et2Cit,Na3Cit,and PFA can all reduce MOVAS calcification induced by high calcium and reduce extracellular calcified plaque and calcium deposition.These inhibitors can inhibit the transdifferentiation of MOVAS to osteoblast-like cells,resulting in a decrease in the activity of alkaline phosphatase(ALP),a marker transformed to osteogenic differentiation.Their inhibitory effects exhibited concentration dependence.The inhibitory effects of the inhibitors at the same concentra-tion showed the following trend:PFA>Na3Cit>Et2Cit.Low concentrations of Na3Cit and Et2Cit inhibit calcification by reducing cell apoptosis,but high concentrations of Na3Cit,Et2Cit,and PFA increased cell apoptosis due to their own toxicity.Et2Cit and Na3Cit as blood anticoagulants can effectively inhibit MOVAS calcification induced by high calcium.

关 键 词：生物矿化 磷酸钙 血管钙化 高钙 柠檬酸二乙酯 柠檬酸钠 膦甲酸钠 

分 类 号：R543[医药卫生—心血管疾病] O614.23[医药卫生—内科学]