Metformin promotes angiogenesis and functional recovery in aged mice after spinal cord injury by adenosine monophosphate-activated protein kinase/endothelial nitric oxide synthase pathway

Treatment with metformin can lead to the recovery of pleiotropic biological activities after spinal cord injury.However,its effect on spinal cord injury in aged mice remains unclear.Considering the essential role of angiogenesis during the regeneration process,we hypothesized that metformin activates the adenosine monophosphate-activated protein kinase/endothelial nitric oxide synthase pathway in endothelial cells,thereby promoting microvascular regeneration in aged mice after spinal cord injury.In this study,we established young and aged mouse models of contusive spinal cord injury using a modified Allen method.We found that aging hindered the recovery of neurological function and the formation of blood vessels in the spinal cord.Treatment with metformin promoted spinal cord microvascular endothelial cell migration and blood vessel formation in vitro.Furthermore,intraperitoneal injection of metformin in an in vivo model promoted endothelial cell proliferation and increased the density of new blood vessels in the spinal cord,thereby improving neurological function.The role of metformin was reversed by compound C,an adenosine monophosphate-activated protein kinase inhibitor,both in vivo and in vitro,suggesting that the adenosine monophosphate-activated protein kinase/endothelial nitric oxide synthase pathway likely regulates metformin-mediated angiogenesis after spinal cord injury.These findings suggest that metformin promotes vascular regeneration in the injured spinal cord by activating the adenosine monophosphate-activated protein kinase/endothelial nitric oxide synthase pathway,thereby improving the neurological function of aged mice after spinal cord injury.

Bone Density and Mechanical Properties in Femoral Bone of Swim Loaded Aged Mice

Effects of swirnming on bone density and mechanical properties of femur were investigated in aged male and female mice. R/1 strain of senescence accelerated mouse (SAM) at eleven months old was used. Two groups of males and two groups of females each consisting of 7 mice were used. One male and one female groups were loaded with a swim regiment of 40 min a day, 5 days a week for 6 consecutive weeks. The remaining groups were used as the controls. All mice were fed with the standard diet and water ad libitum during the experiments.The results of this study indicated that (i) the hady weight was significantly (P<0.05) lower in the swimming groups than in the control groups in boh sexes. (ii) The bone density was significantly higher (P <0.05) in the swimming groups than in the control groups in boh sexes. However, there was no sighficant difference in cortical thickness index. (iii) In the mechanical properties of bone, there were no significant differences in the level of the maximum breaking force, the ultimate stress and the deformation between the swimndng and the contro groups in beth sexes. However, the elasticity of the bone of the female hoce in the swimming group was significantly higher (P<0.05) than that of the control group.These results suggest that regimented swimming for the aged mice might suppress age-associated bone loss, and the effect of exercise in the females is greater that in the males.

Tibolone modulates neuronal plasticity through regulating Tau, GSK3β/Akt/PI3K pathway and CDK5 p35/p25 complexes in the hippocampus of aged male mice

Aging is a key risk factor for cognitive decline and age-related neurodegenerative disorders. Also, an age-related decrease in sex steroid hormones may have a negative impact on the formation of neurofibrillary tangles （NFTs）; these hormones can regulate Tau phosphorylation and the principal kinase GSK3β involved in this process. Hormone replacement therapy decreases NFTs, but it increases the risk of some types of cancer. However, other synthetic hormones such as tibolone （TIB） have been used for hormone replacement therapy. The aim of this work was to evaluate the long-term effects of TIB （0.01 mg/kg and 1mg/kg, intragastrically for 12 weeks） on the content of total and hyperphosphorylated Tau （PHF-1） proteins and the regulation of GSK3β/Akt/PI3K pathway and CDK5/p35/p25 complexes in the hippocampus of aged male mice. We observed that the content of PHF-1 decreased with TIB administration. In contrast, no changes were observed in the active form of GSK3β or PI3K. TIB decreased the expression of the total and phosphorylated form of Akt while increased that of p110 and p85. The content of CDK5 was differentially modified with TIB： it was increased at low doses and decreased at high doses. When we analyzed the content of CDK5 activators, an increase was found on p35; however, the content of p25 decreased with administration of low dose of TIB. Our results suggest a possible mechanism of action of TIB in the hippocampus of aged male mice. Through the regulation of Tau and GSK3β/Akt/PI3K pathway, and CDK5/p35/p25 complexes, TIB may modulate neuronal plasticity and regulate learning and memory processes.

CpG ODN Enhances Immunization Effects of Hepatitis B Vaccine in Aged Mice

Oligodeoxynucleotides(ODN)containing unmethylated CpG dinucleotides in contexts of unique sequence (CpG motifs)is active as adjuvant in induction of cellular and humoral immune responses in young mice.To date,there are only limited reports about effect of CpG ODN on immune responses against hepatitis B(HB) infection in aged mice.Our studies demonstrated there were significant increases in secreting of total anti-HB IgG,IgG1 and IgG2a,as well as of IL-12 and IFN-γ,when CpG ODNs were injected together with hepatitis B antigen in aged mice.Moreover,CpG ODN could stimulate proliferation of spleen lymphocytes in a dose-dependent manner.Taken together,the results we obtained indicate that the adding of CpG ODN into the vaccine antigen might be useful in development of more effective vaccination for inducing anti-HB virus responses in the elderly.Cellular & Molecular Immunology.2004;1(2):148-152.

The effect of adjuvants on vaccine-induced antibody response against influenza in aged mice

While influenza remains a major threat to public health, researchers continue to search for a universal solution to improving the efficacy of the influenza vaccine. Even though influenza affects people of all different ages, it can be extremely hazardous to people of 65 years of age or older since that is the population that makes up the high majority of the death toll caused by influenza-related diseases. Elderly individuals suffer the effects of immunosenescence as they age, which is the diminishing of the overall immune response. Immunosenescence occurs by specifically affecting the adaptive immune response which controls the establishment of immunity after vaccination or infection. There are many studies under way that are trying to find a resolution to the problem of the influenza vaccine not providing enough protection in the elderly population. One of the possible strategies is to seek the use of an optimal adjuvant, an immunological agent that can enhance immune responses, with the current vaccine formulation. Here, we used the murine model to review the effects of adjuvants on the antibody response to influenza vaccines in aged mice. Since adjuvants can enhance the production of important inflammatory cytokines and activation of dendritic cells, the stimulation of these cells are boosted to increase the effectiveness of the influenza vaccine in aged mice which would hopefully translate to the elderly.

Application of a real-ambient fine particulate matter exposure system on different animal models

Simulation of fine particulate matter(PM_(2.5))exposure is essential for evaluating adverse health effects.In this work,an ambient exposure system that mimicked real atmospheric conditions was installed in Taiyuan,China to study impacts of chronic PM_(2.5) exposure on adult and aged mice as well as Sirtuin3 knockout(Sirt3 KO)mice and wild-type(WT)mice.The real-ambient exposure system eliminated the possible artificial effects caused from exposure experiments and maintained the physiochemical characteristics of PM_(2.5).The case studies indicated that aged mice exhibited apparent heart dysfunction involving in-creased heart rate and decreased blood pressure after 17-week of real-ambient PM_(2.5) exposure.Meanwhile,15-week of real-ambient PM_(2.5) exposure decreased the heart rate and amounts of associated catecholamines to induce heart failure in Sirt3 KO mice.Additionally,the increased pro-inflammatory cytokines and decreased platelet related indices suggested that inflammation occurred.The changes of biomarkers detected by targeted metabolomics confirmed metabolic disorder in WT and Sirt3 KO mice after exposed to real-ambient PM_(2.5).These results indicated that the real-ambient PM_(2.5) exposure system could evaluate the risks of certain diseases associated with air pollution and have great potential for support-ing the investigations of PM_(2.5) effects on other types of rodent models.