Effects of CdCl2 and Cd-metallothionein on cultured mesangial cells

Cadmium is a potent nephrotoxin known to cause damage to the proximal tubular epithelium in vivo. The renal glomerulus is less frequently a target and is sensitive to Cd in vitro. We have previously presented evidence that the mesangial cell is a major target for Cd2+ toxicity in the isolated glomerulus (D. M. Templeton and N. Chaitu, Toxicology 61, 119-133, 1990). The present study was undertaken to investigate the sensitivity to Cd of rat mesangial cells grown in homogeneous culture. At a concentration of 1 microM, Cd2+ was a potent inducer of its binding protein metallothionein (MT). Cd2+ inhibited DNA synthesis in these cells with an EC50 of 5.4 +/- 0.4 microM, while preinduction of MT with Zn2+ was protective, raising the EC50 to 17.6 +/- 0.7 microM Cd2+. DNA synthesis in these cells is especially sensitive to Cd2+; only at concentrations of 20 microM Cd2+ and higher were significant effects on cell viability, attachment, and protein synthesis observed. Renal function depends in part on synthesis of specialized matrices by glomerular cells. Synthesis of both matrix and secreted proteoglycans was specifically affected by Cd2+ with an EC50 of about 10 microM for proteoglycan sulfation. We also investigated the effects of Cd-MT on these parameters. Contrary to observations that extracellular Cd-MT is a potent nephrotoxin in vivo, we were unable to demonstrate any effects of Cd-MT on DNA and protein synthesis at Cd concentrations below 60 microM in the cultured cells. Nor did Cd-MT at these concentrations affect DNA or protein synthesis in LLC-PK1 cells, a proximal tubule cell line. This was not due to failure of the cells to take up Cd because they accumulated comparable amounts of Cd whether it was provided as CdCl2 or Cd-MT. We conclude that ionic Cd2+ is the most toxic form of this metal to cultured mesangial cells. While these cells respond to micromolar concentrations of Cd2+ by increasing their content of metallothionein, presumably a protective response, only slightly higher levels may impair the regenerative capacity of mesangial cells, in addition to interfering with the specialized function of matrix synthesis.