Institute of Biomineralization and Lithiasis Research, Jinan University, Guangzhou 510632, China.
Department of Urology, Guangzhou Institute of Urology, Guangdong Key Laboratory of Urology, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou Medical University, Guangzhou, Guangdong 510230, China.
Biomater Adv. 2022 Jun;137:212854. doi: 10.1016/j.bioadv.2022.212854. Epub 2022 May 13.
Injury of renal tubular epithelial cells (HK-2) is an important cause of kidney stone formation. In this article, the repairing effect of polysaccharide (PCP0) extracted from the traditional Chinese medicine Poria cocos and its carboxymethylated derivatives on damaged HK-2 cells was studied, and the differences in adhesion and endocytosis of the cells to nanometer calcium oxalate monohydrate (COM) before and after repair were explored.
Sodium oxalate (2.8 mmol/L) was used to damage HK-2 cells to establish a damage model, and then Poria cocos polysaccharides (PCPs) with different carboxyl (COOH) contents were used to repair the damaged cells. The changes in the biochemical indicators of the cells before and after the repair and the changes in the ability to adhere to and internalize nano-COM were detected.
The natural PCPs (PCP0, COOH content = 2.56%) were carboxymethylated, and three carboxylated modified Poria cocos with 7.48% (PCP1), 12.07% (PCP2), and 17.18% (PCP3) COOH contents were obtained. PCPs could repair the damaged HK-2 cells, and the cell viability was enhanced after repair. The cell morphology was gradually repaired, the proliferation and healing rate were increased. The ROS production was reduced, and the polarity of the mitochondrial membrane potential was restored. The level of intracellular Ca ions decreased, and the autophagy response was weakened.
The cells repaired by PCPs inhibited the adhesion to nano-COM and simultaneously promoted the endocytosis of nano-COM. The endocytic crystals mainly accumulated in the lysosome. Inhibiting adhesion and increasing endocytosis could reduce the nucleation, growth, and aggregation of cell surface crystals, thereby inhibiting the formation of kidney stones. With the increase of COOH content in PCPs, its ability to repair damaged cells, inhibit crystal adhesion, and promote crystal endocytosis all increased, that is, PCP3 with the highest COOH content showed the best ability to inhibit stone formation.
肾小管上皮细胞(HK-2)损伤是结石形成的重要原因。本文研究了从中药茯苓中提取的多糖(PCP0)及其羧甲基化衍生物对损伤的 HK-2 细胞的修复作用,并探讨了修复前后细胞对纳米草酸钙一水合物(COM)的黏附和内吞作用的差异。
用草酸钠(2.8mmol/L)损伤 HK-2 细胞建立损伤模型,然后用不同羧基(COOH)含量的茯苓多糖(PCPs)修复损伤细胞。检测修复前后细胞生化指标的变化及纳米-COM 黏附和内吞能力的变化。
天然 PCPs(PCP0,COOH 含量=2.56%)进行羧甲基化,得到 3 种羧基含量分别为 7.48%(PCP1)、12.07%(PCP2)和 17.18%(PCP3)的羧甲基化茯苓多糖。PCPs 能修复损伤的 HK-2 细胞,修复后细胞活力增强。细胞形态逐渐修复,增殖和愈合率提高。ROS 产生减少,线粒体膜电位极性恢复。细胞内 Ca 离子水平降低,自噬反应减弱。
PCPs 修复的细胞抑制纳米-COM 的黏附,同时促进纳米-COM 的内吞。内吞晶体主要在溶酶体中积累。抑制黏附和增加内吞可以减少细胞表面晶体的成核、生长和聚集,从而抑制结石的形成。随着 PCPs 中 COOH 含量的增加,其修复损伤细胞、抑制晶体黏附、促进晶体内吞的能力均增加,即 COOH 含量最高的 PCP3 表现出抑制结石形成的最佳能力。