Search

Purpurin, an anthraquinone, has potent anti-oxidant and anti-inflammatory effects in various types of brain damage. In a previous study, we showed that purpurin exerts neuroprotective effects against oxidative and ischemic damage by reducing pro-inflammatory cytokines. In the present study, we investigated the effects of purpurin against D-galactose-induced aging phenotypes in mice. Exposure to 100 mM D-galactose significantly decreased cell viability in HT22 cells, and purpurin treatment significantly ameliorated the reduction of cell viability, formation of reactive oxygen species, and lipid peroxidation in a concentration-dependent manner. Treatment with 6 mg/kg purpurin significantly improved D-galactose-induced memory impairment in the Morris water maze test in C57BL/6 mice and alleviated the reduction of proliferating cells and neuroblasts in the subgranular zone of the dentate gyrus. In addition, purpurin treatment significantly mitigated D-galactose-induced changes of microglial morphology in the mouse hippocampus and the release of pro-inflammatory cytokines such as interleukin-1β, interleukin-6, and tumor necrosis factor-α. In addition, purpurin treatment significantly ameliorated D-galactose-induced phosphorylation of c-Jun N-terminal kinase and cleavage of caspase-3 in HT22 cells. These results suggest that purpurin can delay aging by reducing the inflammatory cascade and phosphorylation of the c-Jun N-terminal in the hippocampus., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Response to cerebral ischemia in young animals was very different from that in the adult. The aim of this study was to investigate differences in neuronal death and gliosis in the hippocampal CA1 region (CA1) between adult and young gerbils following 5 and 15 min of transient cerebral ischemia. Delayed neuronal death (DND) of pyramidal cells occurred in the CA1 was similar in all the adult gerbils after 5 and 15 min of ischemia: the DND occurred 4 days after ischemia. In the young groups, DND of pyramidal cells in the CA1 region occurred 7 and 3 days after 5 and 15 min of ischemia, respectively. On the other hand, the activation of GFAP-immunoreactive ((+)) astrocytes and Iba-1(+) microglia was different in the young groups from the adult groups after ischemia. The change pattern of GFAP immunoreactivity in the adult groups was similar in both the adult groups after ischemia; in the young groups, the activation of GFAP(+) astrocytes after 5 min of ischemia was much delayed than that after 15 min of ischemia. Activated Iba-1(+) microglia were aggregated in the stratum pyramidale 4 days after ischemia in all the adult ischemia-operated groups; in the young groups, activated Iba-1(+) microglia were aggregated in the stratum pyramidale 7 days after 5 min of ischemia and 3 days after 15 min of ischemia. These observations indicate that DND in young animals is very different from the adult according to different duration of transient cerebral ischemia and glial activation is very different in young animals after different duration of transient ischemia., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Aging is an inevitable process that occurs in the whole body system accompanying with many functional and morphological changes. Inflammation is known as one of age-related factors, and inflammatory changes could enhance mortality risk. In this study, we compared immunoreactivities of inflammatory cytokines, such as interleukin (IL)-2 (a pro-inflammatory cytokine), its receptor (IL-2R), IL-4 (an anti-inflammatory cytokine), and its receptor (IL-4R) in the cervical and lumbar spinal cord of young adult (2-3 years old) and aged (10-12 years old) beagle dogs using immunohistochemistry and western blotting. IL-2 and IL-2R-immunoreactive nerve cells were found throughout the gray matter of the cervical and lumbar spinal cord of young adult and aged dogs. In the spinal cord neurons of the aged dog, immunoreactivity and protein levels were apparently increased compared with those in the young adult dog. Change patterns of IL-4- and IL-4R-immunoreactive cells and their protein levels were also similar to those in IL-2 and IL-2R; however, IL-4 and IL-4R immunoreactivity in the periphery of the neuronal cytoplasm in the aged dog was much stronger than that in the young adult dog. These results indicate that the increase of inflammatory cytokines and their receptors in the aged spinal cord might be related to maintaining a balance of inflammatory reaction in the spinal cord during normal aging.

The activation of caspase-3 is considered to be a reliable marker for apoptotic cell death, and a 120-kDa fragment of αII-spectrin is generated by caspase-3 mediated cleavage of this structural protein. In the present study, we compared cleaved αII-spectrin (120-kDa) and cleaved caspase-3-immunoreactive cells and their protein levels in the cervical (C₅-C₆) and lumbar (L₃-L₄) levels of the spinal cord in adult (1-2 year-old) and aged (10-12 year-old) dogs (German shepherds). Weak cleaved αII-spectrin and cleaved caspase-3 immunoreactivity was found in neurons of the adult group; however, their immunoreactivity was distinctively increased in the neuronal cytoplasm in the aged group compared to those in the adult group, although the distribution pattern of their neurons was similar between the adult and age group. In addition, cleaved αII-spectrin and cleaved caspase-3 levels in the aged spinal cord were markedly increased compared to those in the adult group. These findings suggest that the increases of cleaved αII-spectrin and cleaved caspase-3 immunoreactivity may be related to aging of the spinal cord in dogs.

Microglia are main form of active immune defense, and they are constantly moving and analyzing the CNS for damaged neurons and infectious agents. In this study, we compared microglia in the spinal cord of the young adult (1-2 years old) and aged (10-12 years old) German Shepherd dogs via immunohistochemistry and western blot analysis for ionized calcium-binding adapter molecule 1 (Iba-1), a microglial marker. In addition, we also observed the interferon-gamma (IFN-gamma), a pro-inflammatory cytokine, and interleukin-1beta (IL-1beta), produced by activated microglia/macrophage, protein levels in these groups. At first, we found that neuronal nuclei (NeuN, a neuronal marker)-immunoreactive neurons were distributed throughout the grey mate of the spinal cord, and there were no significant differences between the adult and aged groups. Most of Iba-1-immunoreactive microglia were morphologically ramified microglia (resting form) in the adult group, while some Iba-1-immunoreactive microglia were morphologically activated microglia in the aged group. In western blot analysis, Iba-1, IFN-gamma and IL-1beta expression were increased in the aged group. This result may be associated with age-dependent changes in the spinal cord.

Myb genes are a family of transcription factors and have been implicated in the control of the proliferation and differentiation of normal and transformed cells. c-Myb is the best characterized member of the myb family. In the present study, we investigated age-dependent changes of c-myb immunoreactivity, its protein and mRNA level in the hippocampus proper (CA1-3 regions) at various age stages in gerbils. In the postnatal month 1 (PM 1) group, c-myb immunoreactivity was detected in non-pyramidal neurons of the strata oriens and radiatum as well as in pyramidal neurons of the stratum pyramidale. At PM 3, c-myb immunoreactivity and its protein level were similar to those at PM 1. Thereafter, c-myb immunoreactivity and its protein level were increased with time. In the PM 24 group, c-myb immunoreactivity, its protein and mRNA levels were highest. These results suggest that the significant increase of c-myb immunoreactivity, protein and mRNA levels in the aged hippocampus may be associated with neuronal aging.