Renal protection by 3H-1,2-dithiole-3-thione against cisplatin through the Nrf2-antioxidant pathway.

Cisplatin is commonly used for the treatment of several solid tumors. However, its clinical use is often limited by renal toxicity. The indirect antioxidant 3H-1,2-dithiole-3-thione (D3T) has been known to protect cells from oxidative damage by up-regulating the expression of antioxidative genes through the transcription factor NF-E2-related factor 2 (Nrf2) pathway. We hypothesized that D3T treatment may be protective against cisplatin-induced nephrotoxicity by enhancing the antioxidative capacity of renal cells. In cultured murine tubular epithelial cells, D3T facilitates the nuclear accumulation of Nrf2 and the subsequent expression of its target genes such as glutamate cysteine ligase (GCL). Increased GSH pool in D3T-treated renal cells appears to be associated with amelioration of cisplatin-mediated cell death. Protective effects of D3T were also observed in mice. Oral administration of D3T (0.25mmol/kg) increased the expression of GCL in mouse kidney, which resulted in suppression of cisplatin-mediated increases in blood urea nitrogen and serum creatinine. Histopathological changes representing cisplatin-induced acute renal failure were also effectively ameliorated by D3T treatment. Collectively, these results indicate that pharmacological activation of the Nrf2 pathway might have a beneficial effect on reducing chemotherapy-associated cytotoxic adverse effects.

Adaptive response to GSH depletion and resistance to L-buthionine-(S,R)-sulfoximine: involvement of Nrf2 activation.

Pharmacological depletion of L-gamma-glutamyl-L-cysteinyl-glycine (GSH) has been implicated in the sensitization of cancer cells to alkylating agents and apoptosis. However, some types of cells do not induce apoptotic response following chemical depletion of GSH. In the present study, we report that murine embryonic fibroblasts (MEFs) can survive in the presence of GSH inhibitor L-buthionine-(S,R)-sulfoximine (BSO), even though most intracellular GSH was depleted. As a cellular adaptive mechanism, BSO treatment effectively activated the NF-E2-related factor 2 (Nrf2) pathway, which led to up-regulation of antioxidant enzymes in these cells through the extracellular signal-regulated kinase cascade. While nrf2-deficient MEFs lost the inducibility of antioxidant genes, which resulted in higher levels of reactive oxygen species accumulation, caspase-3 activation, and cell death than wild-type cells. Finally, nrf2-deficient cells can be more sensitized to doxorubicin-induced cell death by BSO pre-incubation, while wild-type cells were not. In addition, BSO-mediated cell death was facilitated by administering Nrf2 siRNA to chemoresistant human ovarian cancer cells. These results indicate that Nrf2 is the primary factor inducing the cell survival system under GSH depletion and that the effect of BSO as a chemosensitizer might be enhanced by inhibition of Nrf2.

Role of the Nrf2-antioxidant system in cytotoxicity mediated by anticancer cisplatin: implication to cancer cell resistance.

The treatment of alkylating cytotoxic drug cisplatin is often limited by high incidence rate of resistance. In the present study, the potential involvement of the transcription factor Nrf2 in determination of cisplatin cytotoxicity has been investigated. Nrf2-deficient murine embryonic fibroblasts showed increased cell death, cytotoxicity, and apoptosis in response to cisplatin treatment compared to wild-type cells. Cisplatin-resistant human ovarian cancer SK-OV cells, which are retaining 25-fold higher levels of GSH than murine fibroblasts, could be sensitized by inhibition of Nrf2. Transfection with Nrf2 siRNA into SK-OV cells resulted in severe degree of GSH depletion and exacerbated cytotoxicity following cisplatin treatment compared to scrambled RNA control. In conclusion, we propose that the Nrf2 pathway, which plays a protective role in normal cells, can be a potential target to control cancer cell resistance to oxidants, cytotoxic chemicals, and radiation.

Effects of defatted safflower seed extract and phenolic compounds in diet on plasma and liver lipid in ovariectomized rats fed high-cholesterol diets.

Six polyphenolic compounds were isolated from ethylacetate extract secondary to 80% ethanol extraction of defatted safflower seeds. They were categorized into three types: lignans, flavones and serotonin derivatives. Female Sprague-Dawley rats weighing 163.4 +/- 6.3 g were ovariectomized (Ovx) and fed either ethylacetate extract at a level of 1% (w/w) or three types of safflower polyphenolic compounds at a level of 200 mg/kg in a diet containing 0.5% (w/w) cholesterol for four wk. The sham and Ovx control groups were fed the same diet without safflower components. Plasma GOT and GPT levels did not differ among the six experimental groups. The plasma levels of total cholesterol were reduced in the four safflower groups by 20-30% as compared to the Ovx control. The plasma level of HDL-cholesterol was higher in the Ovx+ethylacetate extract group or appeared to be in the three Ovx+safflower polyphenolic groups than in the Ovx control. The level of plasma triglyceride was also significantly lower in the Ovx+lignan group than in the Ovx control. The liver level of cholesterol was significantly reduced in the Ovx+ethylacetate extract group. Fecal excretion of cholesterol increased by the safflower lignans and flavones, whereas that of bile acid was not significantly changed by the safflower polyphenols. Matairesinol and acacetin isolated from safflower seeds reduced the cholesterol content in cultured HepG2 cells at a concentration of 0.01-0.1 microM and all three safflower polyphenolics decreased triglyceride content at the concentration of 0.1 microM. These results suggest that safflower polyphenols have the effect of improving blood lipid status via increasing HDL-cholesterol formation and cholesterol excretion without significant uterotropic action in estrogen-deficient animals.