The purpose of this study was to clarify the effect of silica-induced cytosolic free calcium mobilization and cell injury in immortalized cell lines from transgenic mice kidney harboring SV40 T-antigen gene. The proximal convoluted tubule (S
1)- and the inner medullary collecting tubule (IMCT)- originated cell lines were used. Cytosolic free calcium concentration ([Ca
2+]
i) was measured employing Fura-2 fluorescence and cell injury was evaluated by a vital dye exclusion procedure. Silica increased [Ca
2+]
i in a concentration-dependent manner in S
1 (60μg/ml-600μg/ml) and IMCT (6μg/ml-600μg/ ml). Silica caused a biphasic increase in [Ca
2+]
i which was composed of an initial rapid rise and following sustained phase. Ca
2+ removal from the medium resulted in abolishment of initial and sustained phase of silica (600μg/ml)-induced [Ca
2+]
i in both cell lines. Silica-induced cell injury was increased in a dose-dependent manner. This silica-induced cell injury was attenuated by the pretreatment with EGTA (100μM) and nifedipine (1μM). Cellular ATP content ([ATP]
i) by silica also decreased in a concentration-dependent manner. The relationship between [Ca
2+]
i and [ATP]
i showed that [ATP]
i depletion caused [Ca
2+]
i to rise. This study suggests hat 1) the elevation of [Ca
2+]
i caused by silica is due mainly to influx through plasma membrane Ca
2+ channel and non specific membrane damage (at high concentration) and 2) nephrotoxicity of silica shows site-specificity within the kidney.
View full abstract