Renal injury evokes tubular cholesterol accumulation, mediated in part by increased HMG CoA reductase (HMGCR) levels. The present study was undertaken to define potential molecular determinants of these changes and to ascertain the relative importance of increased cholesterol production versus mevalonate pathway-driven protein prenylation, on the emergence of the so-called postrenal injury "cytoresistant state." Cultured proximal tubule (HK-2) cells were subjected to Fe or ATP depletion injury, followed 1 to 24 hours later by assessments of: 1) sterol transcription factor expression (SREBP)-1 and -2); 2) HMGCR mRNA levels; and 3) Ras/Rho prenylation. HMGCR mRNA and Ras/Rho prenylation were also assessed after in vivo ischemic and Fe-mediated renal damage. Using specific inhibitors, the relative importance of protein prenylation versus terminal cholesterol synthesis on HK-2 cell susceptibility to injury was also assessed. Acute injury induced HK-2 cell SREBP disruption and reductions in HMGCR mRNA. Renal cortical HMGCR mRNA also fell in response to either in vivo ischemic or Fe-mediated oxidant damage. At 24 hours after in vitro/in vivo injury, a time of cholesterol buildup, no increase in Ras/Rho prenylation was observed. Prenylation inhibitors did not sensitize HK-2 cells to injury. Conversely, squalene synthase (terminal cholesterol synthesis) blockade sensitized HK-2 cells to both Fe and ATP depletion attack. We concluded that: 1) acute tubular cell injury can destroy SREBPs and lower HMGCR mRNA. This suggests that posttranscriptional/translational events are responsible for HMGCR enzyme and cholesterol accumulation after renal damage. 2) Injury-induced cholesterol accumulation appears dissociated from increased protein prenylation. 3) Cholesterol accumulation, per se, seems to be the dominant mechanism by which the mevalonate pathway contributes to the postrenal injury cytoresistant state.

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Natural killer (NK) cells are a subset of lymphocytes that are capable of killing tumor cells, virally infected cells and antibody coated cells. Tributyltin (TBT) is a toxic chemical used for various industrial purposes such as: slime control in paper mills, disinfection of circulating industrial cooling waters, anti-fouling agents, and the preservation of wood. TBT can be found in edible items such as fish. A previous study showed that a 1 h exposure of NK cells to TBT caused persistent inhibition of NK-cell ability to destroy tumor cells in the 24 and 48 h periods following exposure and that this loss of function could be significantly prevented and/or reversed if the NK-stimulatory interleukins (IL) 2 or 12 were present during the 24 and 48 h periods. We had also shown that TBT exposure was able to significantly decrease the protein and mRNA levels of the cytotoxic proteins, granzyme B and perforin, and the phosphorylation of cAMP-response-element-binding protein (CREB) under these conditions. In this study we address the effects of IL-2 and IL-12 on the TBT-induced decreases in NK-cell levels of the cytotoxic proteins, their mRNAs, and CREB phosphorylation. IL-2 appeared to prevent/reverse TBT-induced declines in perforin protein levels and the mRNA for perforin seen in the 24 h period following a 1 h exposure to 300 nM TBT. However, the TBT-induced decreases in the levels of perforin and perforin mRNA seen in the 48 h period following a 1 h exposure to TBT were not statistically significantly prevented/reversed by IL-2. Additionally, the TBT-induced decreases in granzyme B, granzyme B mRNA, and CREB phosphorylation were not statistically significantly reversed by either IL-2 or IL-12 after 24 or 48 h.