Mutation Research/DNAging最新文献

DNA polymerase α (pol α) purified from human diploid fibroblasts (HDF) and from livers of C57BL/6N mice showed age-related decreases in: (1) mRNA levels; (2) the amount of enzyme isolated per cell; and (3) enzyme activity (HDF); as well as: a) the amount of enzyme isolated; b) the specific activity; and c) the enzyme fidelity (liver). Hepatic pol α from dietary restricted (DR) mice exhibited less of a decline in specific activity and copied synthetic DNA templates with relatively higher fidelity than did enzymes from animals fed ad libitum (AL). Pol α from fetal-derived HDF exhibited increased expression compared with aged donor-derived HDF, with both fetal and old cell pol α in normal cells being expressed at lower levels than in their transformed cell corollaries. Treatment of human pol α from aged donor-derived HDF with a pol α accessory protein isolated from log phase murine cells resulted in increased pol α binding of DNA and increased pol α activity. However, highly active pol α isolated from fetal-derived or transformed HDF, or from transformed murine cells, showed little or no activity enhancement in the presence of accessory protein. These data indicate that, as a function of increased age, there is a decrease in pol α expression and specific activity in HDF, as well as decreases in specific activity and fidelity of pol α in essentially amitotic murine hepatic tissues. Dietary restriction impedes the age-related declines in both activity and fidelity of hepatic pol α in mice. The data further indicate that transformation of slowly dividing HDF is associated with increased expression of pol α, but suggest that increased expression alone is not sufficient to explain the difference in polymerase activity levels between parental and transformed HDF. Lastly, the data suggest that interaction of pol α with an essential accessory protein may be altered as a function of age, an alteration that appears to be correlated with the decline in pol α DNA binding and specific activity.

The basic mechanisms of aging and its retardation by caloric restriction (CR) remain unclear. One suggested means by which CR could retard aging is based on production of mitochondrial free radicals, and efficiency of their subsequent metabolism. Currently, there is little information concerning the influences of age and CR on the rates of in vivo mitochondrial free radical production. However, evidence for CR-induced modulation of free radical detoxification capacities is mounting. The direction of the influence of CR on free radical detoxification is tissue-specific. These effects are broad and appear to provide positive advantage.

The effect of caloric restriction (CR) on xenobiotic metabolizing enzyme activities results in alterations in the metabolic activation of chemical carcinogens, with a resultant impact on DNA-carcinogen adduct formation and DNA repair. Using aflatoxin B1 (AFB1) and benzo[a]pyrene (BP) as model carcinogens, we studied the effect of CR on the metabolic activation of these carcinogens and carcinogen-induced DNA damage and repair in terms of AFB1-DNA and BP-DNA adduct formation and removal. Male Fischer 344 rats fed calorie restricted diets (60% of the food consumption for ad libitum-fed rats) showed a reduction in the metabolic activation of AFB1 and decrease in both the in vitro and in vivo AFB1-DNA adduct formation. However, CR increased the activity of BP metabolizing enzymes resulting in an enhancement of BP-DNA adduct formation. Our results indicate that the effect of CR on metabolic activation of xenobiotics is dependent upon the selected xenobiotic metabolizing enzymes whose activities may be significantly altered by CR, and upon the nature of the chemical carcinogens which exert different structure-activity relationships during the process of chemically induced carcinogenesis.

Caloric restriction in rodents results in increased longevity and a decreased rate of spontaneous and chemically induced neoplasia. The low rates of spontaneous neoplasia and other pathologies have made calorically restricted rodents attractive for use in chronic bioassays. However, caloric restriction also alters hepatic drug metabolizing enzyme (DME) expression and so may also alter the biotransformation rates of test chemicals. These alterations in DME expression may be divided into two types: (1) those that are the direct result of caloric restriction itself and are detectable from shortly after the restriction is initiated; (2) those which are the result of pathological conditions that are delayed by caloric restriction. These latter alterations do not usually become apparent until late in the life of the organism. In rats, the largest direct effect of caloric restriction on liver DMEs is an apparent de-differentiation of sex-specific enzyme expression. This includes a 40–70% decrease in cytochrome P450 2C11 (CYP2C11) expression in males and a 20–30% reduction of corticosterone sulfotransferase activity in females. Changes in DME activities that occur late in life in calorically restricted rats include a stimulation of CYP2E1-dependent 4-nitrophenol hydroxylase activity and a delay in the disappearance of male-specific enzyme activities in senescent males. It is probable that altered DME expression is associated with altered metabolic activation of chemical carcinogens. For example the relative expression of hepatic CYP2C11 in ad libitum-fed or calorically restricted rats of different ages is closely correlated with the amount of genetic damage in 2-acetylaminofluorene- or aflatoxin B₁-pretreated hepatocytes isolated from rats of the same age and caloric intake. This suggests that altered hepatic drug and carcinogen metabolism in calorically restricted rats can influence the carcinogenicity of test chemicals.

Protein restriction (PR) and caloric restriction (CR) similarly impinge upon various physiological factors that can significantly inhibit the growth of DNA-damaged tissue and, therefore, carcinogenesis. Whether this effect is largely, or only in part, due to simple inhibition of body weight gain is examined. Among their many other health-improving effects, PR and CR delay the onset of puberty. It has been suggested that animals have developed mechanisms to cope with lean periods and that, when food is limited, resources are diverted from those physiological functions that offer no benefit for immediate survival (e.g., reproductive capacity) to thereby support an increase in the maintenance functions that prolong life. PR has also been shown to affect numerous other varied mechanisms that can affect carcinogenesis, including gene expression and metabolism of xenobiotics. The effects of PR on initiational and promotional growth of DNA-damaged tissue is also discussed. PR also seems to boost antioxidant defenses and inhibit the accumulation of oxidative damage (as does CR). Protein restricted animals have been shown to accumulate more calories, but develop fewer preneoplastic lesions and tumors than their high-protein counterparts. This observation seems quite counter to most ideas about dietary restrictions and CR. Despite the fact that both PR and CR induce many beneficial physiological effects in common, it is possible that PR is the more feasible option for human consideration. The levels of PR likely to improve health without negative side effects are discussed.

Restriction of diet and macronutrients has been reported to modulate the toxicity of numerous chemical agents. Of the various forms of restriction studied, using nutritionally adequate diets, food restriction (FR) appears to be most effective, but protein restriction (PR), fat restriction (FtR), carbohydrate restriction (CbR), and excess of dietary fiber (FE) also affect toxicity and the spontaneous diseases that define the background incidence in toxicity tests. The heterogeneity of the dietary macronutrients complicates simple analysis of their effects. Additionally, the interrelationships between these various components in the complex dietary mixture often make experiments difficult to interpret.

Despite these complexities, a simple model is presented, which considers the effects of dietary manipulations on the individual steps in the interaction of organism and agent, and puts the varied effects that can occur within an organism into context. Ultimately, many of the effects of dietary modulation on these steps in toxicogenesis can be considered as changing agent exposure and the biologically available dose. The effects of macronutrient restriction are discussed in terms of effects on agent at the interface of organism and toxicant, agent disposition, agent metabolism, and repair of toxicant-induced damage at the level of the genome. After illustrating the influence of these nutritional effects on the chronic bioassay, using mouse liver tumors as an example, the significance of these effects for chronic and short-term testing is discussed. Additionally, methods to address the impact of nutritional factors on toxicity testing are suggested.

Decreased dietary intake of fat and/or calories generally results in a lower incidence of mammary gland tumors in rodents. Feeding of either low-fat or calorie-restricted diets to rats also has been shown to result in decreased levels of oxidative DNA damage. Since oxidative DNA damage is suggested to have a role in carcinogenesis, this may be one mechanism by which dietary change can reduce cancer risk. The effects of calorie-restricted diets on both oxidative DNA damage levels and mammary gland tumor incidence are generally more pronounced than that of low-fat diets. There is, however, some difficulty in defining what amount of fat should be used to prepare ‘low-fat’ and ‘high-fat’ rodent diets as well as what a suitable fat intake for control diets should be in studies that examine the effects of dietary fat and/or calories on tumorigenesis. In particular, the promoting effects of dietary fat may be exerted only up to a certain level of fat, above which no further effect is observed. Another difficulty in the interpretation of the results is that there may be a time-dependent effect of high fat diets on oxidative damage, with increased damage resulting only when the diets are fed for longer periods of time. The appropriate experimental approach to model human dietary exposures therefore remains to be determined. Although the effects of caloric intake on mammary gland tumorigenesis appear to be more pronounced than that of fat intake, low-fat diets still may be useful as a preventive measure in human populations to reduce breast cancer risk for individuals who cannot safely reduce their caloric intake.

Dietary restriction is the only experimental manipulation known to extend lifespan and retard aging in mammals. Therefore, it is a powerful tool for identifying cellular processes that are involved in aging and senescence. Recently, several laboratories have begun to examine the effects of dietary restriction on the integrity of the genome and the ability of cells to repair DNA. In most studies, it was found that the repair of DNA damage, as measured by unscheduled DNA synthesis, was significantly higher in cells isolated from rodents fed calorie-restricted diets compared to cells isolated from rodents fed ad libitum. Dietary restriction also was observed to be associated with a reduction of the levels of certain types of DNA damage; however, preliminary experiments suggest that the effect of dietary restriction on the age-related accumulation of DNA damage depends on the type of DNA damage studied.