高等学校化学学报 ›› 2020, Vol. 41 ›› Issue (6): 1231-1242.doi: 10.7503/cjcu20190713

• 研究论文: 无机化学 • 上一篇    下一篇

不同草酸/钙摩尔比条件下草酸钙晶体的生长及对HK-2细胞的毒性

刘虹1,邹国均1,孙新园2,*(),欧阳健明1,*()   

  1. 1. 暨南大学生物矿化与结石病防治研究所, 广州 510632
    2. 广州医科大学附属第一医院, 广州 510230
  • 收稿日期:2019-12-25 出版日期:2020-06-10 发布日期:2020-03-06
  • 通讯作者: 孙新园,欧阳健明 E-mail:sunxinyuan1985@163.com;toyjm@jnu.edu.cn
  • 基金资助:
    国家自然科学基金(21371077);国家自然科学基金(21975105)

Differences of Growth and Cytotoxicity of Calcium Oxalate Crystals Formed on HK-2 Cells Under Different Oxalic Acid/Calcium Ratios

LIU Hong1,ZOU Guojun1,SUN Xinyuan2,*(),OUYANG Jianming1,*()   

  1. 1. Institute of Biomineralization and Lithiasis Research, Jinan University, Guangzhou 510632, China
    2. The First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510230, China
  • Received:2019-12-25 Online:2020-06-10 Published:2020-03-06
  • Contact: Xinyuan SUN,Jianming OUYANG E-mail:sunxinyuan1985@163.com;toyjm@jnu.edu.cn
  • Supported by:
    † National Natural Science Foundation of China(21371077);National Natural Science Foundation of China(21975105)

摘要:

研究了不同草酸/钙(Ox/Ca)摩尔比对CaOx晶体在损伤前后的人肾近曲小管上皮细胞(HK-2)表面的生长差异及形成的晶体对细胞的毒性差异. 实验结果表明, CaOx过饱和溶液对正常细胞和损伤细胞均会产生进一步的损伤, 导致细胞活力、 溶酶体的完整性和线粒体膜电位降低, 而细胞内活性氧(ROS)、 细胞骨架的紊乱程度、 磷酯酰丝氨酸(PS)外翻比例和骨桥蛋白(OPN)表达量均增加; 且随着过饱和溶液中Ox/Ca摩尔比的增加而损伤加重. 正常细胞主要诱导二水草酸钙(COD)晶体形成, 且COD的含量与Ox/Ca摩尔比成正相关. 损伤细胞表面主要生成一水草酸钙(COM), 且晶体的数量和聚集程度与Ox/Ca摩尔比成正相关. 相比于正常细胞, 损伤细胞诱导的晶体棱角更加尖锐, 其对细胞的损伤大于棱角圆钝的晶体. 实验结果还表明, 降低CaOx的过饱和度、 减小Ox/Ca摩尔比和修复受损伤的肾上皮细胞均有利于抑制CaOx结石形成.

关键词: 草酸钙, Ox/Ca摩尔比, 细胞毒性, 肾结石, 过饱和度

Abstract:

We investigated the growth differences of CaOx crystals with different oxalic acid/calcium(Ox/Ca) molar ratios on the surface of human renal proximal tubule epithelial cells(HK-2) before and after injury and the toxicity of the crystals formed on the cells. CaOx supersaturated solution caused further damage to both the normal and damaged cells, resulting in the decrease of cell viability, lysosomal integrity and mitochondrial membrane potential, while the increase of intracellular reactive oxygen species(ROS), cytoskeleton disorder, phosphatidylserine(PS) eversion ratio and osteopontin(OPN) expression. With the increase of Ox/Ca molar ratio in saturated solution, the cell damage increased. Normal cells mainly induce the formation of calcium oxalate dihydrate(COD) crystals, and the percentage of COD is positively correlated with the Ox/Ca ratio. Damaged cells mainly induced calcium oxalate monohydrate(COM), and the number and aggregation of crystals are positively correlated with Ox/Ca molar ratio. Compared with the control group cells, the crystal edges induced by the damaged group cells are sharper, and the damage of sharp crystals to the cells is greater than that of round and blunt crystals. The results of this study suggest that reducing the supersaturation of CaOx, reducing the Ox/Ca molar ratio and repairing the damaged renal epithelial cells are all beneficial to inhibit CaOx stone formation.

Key words: Calcium oxalate, Ox/Ca molar ratio, Cytotoxicity, Kidney stones, Supersaturation degree

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