HKC细胞损伤前后对草酸钙晶体吞噬能力的差异

采用人类肾脏近端小管上皮细胞系(HKC)建立氧化性损伤模型,研究损伤前后HKC调控草酸钙(CaOxa)结晶的差异。采用CCK-8法检测HKC的细胞活性;利用激光共聚焦显微镜观察HKC损伤后表达的晶体粘附分子骨桥蛋白(OPN);采用倒置显微镜观察HKC的形态变化;采用扫描电子显微镜(SEM)观察HKC微结构及其诱导的晶体;采用X射线衍射分析(XRD)表征晶体的组分。在CaOxa过饱和溶液中,正常HKC主要诱导形成二水草酸钙(COD)晶体,而损伤HKC则同时诱导了COD和一水草酸钙(COM)晶体。正常HKC对COD晶体有较强的吞噬能力,而损伤HKC的这种能力较弱;HKC损伤后表达OPN,促进CaOxa晶体的成核和聚集,从而增加了肾结石形成的危险性。

Difference of Endocytosis to Calcium Oxalate Crystals by HKC Cells before and after Injury

An oxidative injury model of human kidney proximal tubule epithelial cell line (HKC) was used to investigate the difference of crystallization of calcium oxalate (CaOxa) modulated by HKC before and after injury. Cell viability was examined by using Cell Counting Kit (CCK-8) assay; the change of expression of crystal adherent molecule osteopontin (OPN) on HKC after injury was observed by laser confocal microscope; the morphological change of HKC was observed by inverted microscope; the microstructure of HKC and the crystals induced by HKC were observed by means of scanning electron microscopy (SEM); the crystal component was characterized by X-ray diffraction (XRD). Calcium oxalate dihydrate (COD) crystals were mainly induced from CaOxa supersaturated solution by normal HKC, while COD crystals and calcium oxalate monohydrate (COM) crystals were simultaneously induced by injured HKC. The endocytosis of normal HKC to COD crystals was strong, but weak for injured HKC. After expression of OPN, the injured HKC can promote nucleation and aggregation of CaOxa crystals. It increases the risk of urolithiasis formation.