Kaizhi G. Sharman

Chronic exposure to aluminum in drinking water increases inflammatory parameters selectively in the brain.

A link between aluminum (Al) exposure and age‐related neurological disorders has long been proposed. Although the exact mechanism by which the metal may influence disease processes is unknown, there is evidence that exposure to Al causes an increase in both oxidative stress and inflammatory events. These processes have also been suggested to play a role in Alzheimers disease (AD), and exposure to the metal may contribute to the disorder by potentiating these events. Al lactate (0.01, 0.1, and 1 mM) in drinking water for 10 weeks increased inflammatory processes in the brains of mice. The lowest of these levels is in the range found to increase the prevalence of AD in regions where the concentrations of the metal are elevated in residential drinking water (Flaten [2001] Brain Res. Bull. 55:187–196). Nuclear factor‐κB as well as tumor necrosis factor‐α (TNF‐α) and interleukin 1α (IL‐1α) levels were increased in the brains of treated animals. The mRNA for TNF‐α was also up‐regulated following treatment. Enhancement of glial fibrillary acidic protein levels and reactive microglia was seen in the striatum of Al‐treated animals. The level of amyloid beta (Aβ40) was not significantly altered in the brains of exposed animals. Insofar as no parallel changes were observed in the serum or liver of treated animals, the proinflammatory effects of the metal may be selective to the brain. Al exposure may not be sufficient to cause abnormal production of the principal component of senile plaques directly but does exacerbate underlying events associated with brain aging and thus could contribute to progression of neurodegeneration.

Dietary melatonin selectively reverses age-related changes in cortical cytokine mRNA levels, and their responses to an inflammatory stimulus

The basal levels of expression of mRNA of cytokines, interleukin-6 (IL-6) and tumor necrosis factor (TNF-alpha), in the cerebral cortex of 5 and 26 month-old male B6C3F1 mice have been compared. In addition, the responsivity of animals of differing age to an inflammatory stimulus (lipopolysaccharide, LPS) has been studied. Basal levels of both of these cytokine mRNAs were elevated in aged animals relative to the younger group. However LPS administration led to a robust increase in cytokine mRNA levels in the younger animals but in aged mice, there was either an unchanged (IL-6) or a depressed (TNF-alpha) response. Administration of dietary melatonin (200 ppm) to aged mice for 6 weeks prior to sacrifice, resulted in reduction of basal levels of cytokine mRNA to values found in the younger animals. Furthermore, following administration of LPS to melatonin fed animals, cerebral cytokine mRNA levels were significantly elevated rather than being unchanged or depressed. Taken together these findings reflect a trend in the cortices of melatonin-treated aged mice, to more closely approximate the status of younger mice. For comparative purposes, parallel studies were carried out using an immunologically active organ (spleen) and a non-neural organ with a low rate of cell turnover (heart muscle). In both these tissues, basal levels of cytokine mRNAs of animals of either age were very low, and there was a marked positive response to LPS. Dietary melatonin had no effect on the responses of TNF-alpha mRNA to LPS but attenuated the reaction of splenic IL-6 mRNA, thus bringing the response closer to that of the younger mice.

Age‐related changes in murine CNS mRNA gene expression are modulated by dietary melatonin

Abstract:  Brain cellular functions decline with normal aging, accompanied by a changing profile of gene expression. Gene array analysis was used to quantitatively estimate messenger RNA (mRNA) expression levels in the cerebral cortex of both young (4‐month) and old (27‐month) B6C3F1 male mice. A stringent degree of significance was obtained by using multiple gene chips. Out of 12,423 mRNA levels, only 25 changed significantly with age. Nine of these genes coded for inflammatory proteins, all of which were elevated in aged, relative to younger mice. Melatonin (200 p.p.m.) included in the diet of aged animals for 8 wk elevated serum and cortical melatonin and reversed 13 of the 25 genes altered with age. In no case did melatonin potentiate age‐related changes in gene expression. The restoration of a more youthful gene profile to brains of aged animals by melatonin, to a large extent, involves reversal of age‐induced elevation of basal inflammatory parameters.

Reversal of biochemical and behavioral parameters of brain aging by melatonin and acetyl L-carnitine

The potential utility of dietary supplementation in order to prevent some of the oxidative and inflammatory changes occurring in the brain with age, has been studied. The cerebral cortex of 27-month-old male B6C3F1 mice had elevated levels of nitric oxide synthase 1 (EC 1.14.13.39) (nNOS) and peptide nitrotyrosine relative to cortices of younger (4-month-old) animals. After 25-month-old mice received basal diet together with 300 mg/l acetyl L-carnitine in the drinking water for 8 weeks, these levels were fully restored to those found in younger animals. A partial restoration was found when old animals received basal diet supplemented with 200 ppm melatonin in the diet. Levels of mRNA (messenger RNA) for nNOS were unchanged following these treatments implying translational regulation of nNOS activity. Behavioral indices indicative of exploratory behavior were also depressed in aged animals. Dietary supplementation with melatonin or acetyl L-carnitine partially reversed these changes. These findings suggest that dietary supplementation cannot merely arrest but indeed reverse some age-related increases in markers of oxidative and inflammatory events occurring with the cortex.

Effects of melatonin and age on gene expression in mouse CNS using microarray analysis

The expression levels of a number of genes associated with inflammation and immune function change with advancing age. Melatonin modulates gene expression levels of several of these genes. Therefore the declining levels of melatonin associated with age may play a role in the physiological effects of aging. We used oligonucleotide microarrays to measure age-related changes in mRNA expression in the murine CNS, and to study the effect of prolonged administration of dietary melatonin upon these changes. CB6F1 male mice were fed 40 ppm melatonin for 2.1 months prior to sacrifice at age 26.5 months, and compared with both age-matched controls and young, 4.5-month-old untreated controls. Total RNA was extracted from whole brain (excluding cerebellum and brain stem) and individual samples were hybridized to Affymetrix Mouse 430-2.0 arrays. The expression of a substantial number of genes was modulated by melatonin treatment and changes in selected genes were validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR). A subset of these genes did not change with age. Conversely, some genes modulated by age were also modulated by melatonin treatment. In general, melatonin treatment drove the expression levels of these genes closer to the expression levels detected in the younger animals. Notably, the abundance of lipocalin 2 (Lcn2) mRNA increased with age and was decreased in old animals treated with melatonin. Lcn2 is a member of the acute phase response family of proteins and its mRNA levels in the brain increase in response to inflammation. Many of the genes with expression reduced by melatonin are involved in inflammation and the immune system. This suggests that melatonin treatment may influence the inflammatory responses of old animals, driving them to resemble more closely those occurring in young animals.

Melatonin Causes Gene Expression in Aged Animals to Respond to Inflammatory Stimuli in a Manner Differing from that of Young Animals

Groups of younger and aged mice were fed either minimal basal diet or the same diet containing 40 ppm melatonin. After 9.3 weeks half of each of these 4 groups of animals received either an intraperitoneal injection of lipopolysaccharide (LPS) or of saline. Three hours after this treatment, all animals were killed and mRNA from brains extracted. Quantitative PCR was performed on 13 selected mRNA species reflecting various aspects of the inflammatory pathway, the melatonin receptor, and a key glycolytic enzyme. An overall trend observed was that the effect of melatonin in modulating LPS-provoked immune responses differed markedly in old and young animals. Melatonin tended to enhance the reaction of younger animals to LPS but suppressed the inflammatory response of older mice. This difference with aging suggests that key immune processes are markedly altered by aging. It is likely that the ability of the immune system to mount a defense is impaired in older animals.

Extended exposure to dietary melatonin reduces tumor number and size in aged male mice

Several sets of male mice were given dietary melatonin over a series of experiments performed during a nine year period. Overall, melatonin-supplemented mice aged ≥26 months at sacrifice had significantly fewer tumors with lower severity than similarly aged control animals. The studies were originally designed to explore the potential of this agent for reducing the rate of onset of some genetic indices of brain aging. When these animals were sacrificed they were routinely examined for overt evidence of tumors and when these were found, a note was made of their occurrence, and of their size. Tumors are commonly found during senescence of several strains of mice. Since tumorigenesis was not the original intent of the study, these observations were recorded but not pursued in greater detail. In this report, these data have now been collated and summarized. This analysis has the disadvantage that tumor origin and morphology were not recorded. However, the study also has the advantage of being conducted over an extended period of time with many groups of animals. In consequence, many extraneous factors, which could be potential confounders, such as seasonal or dietary variations, are unlikely to have interfered with the analysis. The use of more than one mouse strain strengthens the possibility that the findings may have general relevance. Both aged and young animals were included in the original experiments but the tumor incidence in animals younger than 25 months was very low.

Parallel changes in gene expression in aged human and mouse cortex

The intensity of expression of over 20,000 genes and expressed sequence tags within the cerebral cortex has previously been described for both the human and mouse genomes. In both these species, the degree of expression of a relatively limited number of cortical genes, around 300, is significantly altered during senescence. The extent of similarity between age-related alterations of levels of specific mRNAs in either species has been compared. There is a significant correlation between species in those genes whose expression changes markedly in either direction with aging. This parallel serves to validate the use of mouse strains to study general age-related genetic events associated with aging.

THE HERBAL PRESCRIPTION YOUKONGDAN MODULATES RODENT MEMORY, ISCHEMIC DAMAGE AND CORTICAL mRNA GENE EXPRESSION

The effects of the herbal prescription youkongdan (YKD) on memory performance of rodents following cerebral ischemia/reperfusion and scopolamine administration were measured and compared to the age-related changes in mRNA expression induced by dietary supplementation of YKD. Following ischemia, YKD decreased neuronal cell loss in the CA1 region of rat hippocampus by 89% relative to controls. YKD improved the water maze performance of both ischemic and scopolamine-treated animals. Dietary administration of YKD resulted in significant modulation of Egr1, Grp78, Hsp86, SOD1, and αB crystallin mRNA expression and a trend toward increased exploratory behavior in older mice.