Molecular Mechanism of Adhesion of Monohydrate and Dihydrate Calcium Oxalate Crystals on Injured Kidney Epithelial Cells†

doi:10.7503/cjcu20160035

Abstract

Abstract:

Effects of cell injury on calcium oxalate monohydrate(COM) and calcium oxalate dihydrate(COD) microcrystalline adhesion and cellular response of calcium oxalate microcrystalline on African green monkey renal epithelial(Vero) cells after adhesion were evaluated. COM amd COD crystal adhesion to injured Vero cells increased oxidative damage degree, the LDH release amount, reactive oxygen species(ROS) and dead cells and decreased cell viability. The cells shrinked and apoptotic bodies appeared. COM crystals caused more serious damage to injured Vero cells than COD crystals. The results of scanning electron microscopy(SEM) showed that the adhesive capacity of injured Vero cells to COM was significantly stronger than the control group, which enhanced crystals adhesion and aggregation. Laser scanning confocal microscope showed that Vero cell injury increased the expression of crystal binding hyaluronic acid(HA) molecules which were the most important reasons for promoting microcrystalline adhesion. The microcrystalline adhesion and aggregation on cellular surface were positively correlated to cell injury degree. These findings indicated that cell injury was the leading risk factor of kidney stone formation, which may provide insights into the mechanisms of kidney stone formation from molecular and cellular levels.

Key words: Cell-mediation, Biomineralization, Crystal adhesion, Calcium oxalate, Hyaluronic acid

CLC Number:

O614.23

TrendMD:

GAN Qiongzhi, SUN Xinyuan, YAO Xiuqiong, OUYANG Jianming. Molecular Mechanism of Adhesion of Monohydrate and Dihydrate Calcium Oxalate Crystals on Injured Kidney Epithelial Cells^†[J]. Chem. J. Chinese Universities, 2016, 37(6): 1050.

Figures/Tables 10

Fig.1 SEM images of COM(A) and COD(B) crystals

Fig.2 Cell viability(A) and LDH release(B) after interaction of COM and COD with Vero cells with different damage degree for 6 hH2O2 injury time: 1 h; crystal concentration: 100 μg/mL; interaction time: 6 h.a. Control; b. with micron COD; c. with micron COM.

Fig.3 Observation of cell morphology of Vero cells with different damage degree after interaction with COM and COD crystals for 6 h(A1)—(D1) Control group; (A2)—(D2) with COD; (A3)—(D3) with COM. c(H2O2)/(mmol·L-1): (A1)—(A3) 0; (B1)—(B3) 0.3; (C1)—(C3) 0.5; (D1)—(D3) 1.0. H2O2 injury time: 1 h; crystal concentration: 100 μg/mL.

Fig.4 Fluorescence micrographs of PI staining result after the adhesion of Vero cells with different damage degree to COM and COD (A1)—(D1) Control group; (A2)—(D2) with COD; (A3)—(D3) with COM. c(H2O2)/(mmol·L-1): (A1)—(A3) 0; (B1)—(B3) 0.3; (C1)—(C3) 0.5; (D1)—(D3) 1.0.

Fig.5 PI staining result after the adhesion of Vero cells with different damage degree to COM and COD

Fig.6 Detection of intracellular ROS level of Vero cells after interaction with COM and COD crystals for 6 h(A1)—(D1) Control group; (A2)—(D2) with COD; (A3)—(D3) with COM.c(H2O2)/(mmol·L-1): (A1)—(A3) 0; (B1)—(B3) 0.3; (C1)—(C3) 0.5; (D1)—(D3) 1.0.

Fig.7 HA expression after the adhesion of Vero cells with different damage degree to COM and COD using laser confocal microscope(A1)—(D1) Control group; (A2)—(D2) with COD; (A3)—(D3) with COM.c(H2O2)/(mmol·L-1): (A1)—(A3) 0; (B1)—(B3) 0.3; (C1)—(C3) 0.5; (D1)—(D3) 1.0.

Fig.8 SEM images of COM adhered on Vero cells with different damage degreec(H2O2) /(mmol·L-1): (A) 0; (B) 0.3; (C) 1.0. Crystal concentration: 100 μg/mL; injury time: 1 h; adhesion time: 6 h.

Fig.9 SEM images of COD adhered on Vero cells with different damage degreec(H2O2)/(mmol·L-1): (A) 0; (B) 0.3; (C) 1.0. Crystal concentration: 100 μg/mL; injury time: 1 h; adhesion time: 6 h.

Fig.10 Adhesion amount change of COM and COD on surface of Vero cells with different damage degree

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