• Journal of Animal Reproduction and Biotechnology
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• v.34 no.1
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• pp.35-39
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• 2019

This study aimed to recover the ovarian function through in vitro culture of preantral follicles from aged mice. First, we isolated the preantral follicles from ovaries of sixty-seven-week old B6D2F1 mice with decreased fecundity to know how many follicles were present in them, which was 6 preantral follicles including 2 primary, 2 early secondary and late secondary follicles from 8 aged mice. It was confirmed that a few follicles (~2) were present in aged mice through histological analysis compared to adult mice as control. The 9 days of in vitro culture of preantal follicles showed in vitro growth and induced maturation after treatment with hCG (2.5 IU/mL) and EGF (5 ng/mL). Cumulus cells in the cumulus-oocyte complexes (COCs) were removed using hyaluronidase and oocytes at the germinal vesicle (GV) and GV breakdown (GVBD) were obtained from preantral follicle culture of aged mice in vitro. In conclusion, these observations demonstrated that there still were a few preantral follicles in the ovaries of 67 week-old mice, which we were able to culture in vitro and oocytes were obtained from them. This study proposed an in vitro culture system using preantral follicle as a therapeutic strategy for fertility preservation in humans for assisted reproductive medicine.

• Journal of Embryo Transfer
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• v.33 no.2
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• pp.69-73
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• 2018

This study was conducted to examine the influences of two human chorion gonadotrophins (hCGs) being injected into young or aged (45- to 65-week old) outbred (ICR) mice on developmental capacity of oocytes retrieved. In vitro-culture and parthenogenetic activation of oocytes retrieved were employed for the assessment. Superovulation was determined as being induced when more than 25 oocytes were retrieved. No aged mice were superovulated, while in contrast, 67-100% were superovulated in the 6- to 8-week-old (young) mice. In the aged, hCG injection yielded better retrieval (5 vs. 13 to 14.8 oocytes/mouse). Overall, no significant difference between two hCGs was detected but between the young and aged, significant differences in maturational arrest (0% vs. 39% MI arrest and 46% vs. 15% degeneration) and developmental capacity (24% vs. 46% 8-cell embryo development) were detected. In conclusion, hCG injection contributes to increasing oocyte retrieval from aged outbred mice, but the kinds of gonadotrophin influenced the efficiency of hyperstimulation induction in specific ages.

• Journal of Embryo Transfer
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• v.33 no.4
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• pp.305-311
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• 2018

Gonadotropin releasing hormone (GnRH) centrally plays a role in control of the hypothalamic-pituitary-gonadal axis-related hormone secretions in the reproductive neuroendocrine system. In addition, hormone receptors like luteinizing hormone receptor (LHR) are important element for hormones to take effect in target organ. However, ageing-dependent changes in terms of the distribution of GnRH neurons in the brain and LHR expression in the acyclic ovary have not been fully understood yet. Therefore, we comparatively investigated those ageing-dependent changes using young (1-5 months), middle (11-14 months) and old (21-27 months) aged female mice. Whereas a number of GnRH positive fibers and neurons with monopolar or bipolar morphology were abundantly observed in the brain of the young and middle aged mice, a few GnRH positive neurons with multiple dendrites were observed in the old aged mice. In addition, acyclic ovary without repeated development and degeneration of the follicles was shown in the old aged mice than others. LHR expression was localized in theca cells, granulosa cell, corpora lutea and atretic follicle in the ovaries from young and middle aged mice, in contrast, old aged mice had few positive LHR expression on the follicles due to acyclic ovary. However, the whole protein level of LHR was higher in the ovary of old aged mice than others. These results are expected to be used as an important basis on the relationship between GnRH and LHR in old aged animals as well as in further research for reproduction failure.

• Journal of Ginseng Research
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• v.39 no.3
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• pp.250-256
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• 2015

Background: It has been known that ginseng can be applied as a potential nutraceutical for memory impairment; however, experiments with animals of old age are few. Methods: To determine the memory enhancing effect of red ginseng, C57BL/6 mice (21 mo old) were given experimental diet pellets containing 0.12% red ginseng extract (approximately 200 mg/kg/d) for 3 mo. Young and old mice (4 mo and 21 mo old, respectively) were used as the control group. The effect of red ginseng, which ameliorated memory impairment in aged mice, was quantified using Y-maze test, novel objective test, and Morris water maze. Red ginseng ameliorated age-related declines in learning and memory in older mice. In addition, red ginseng's effect on the induction of inducible nitric oxide synthase and proinflammatory cytokines was investigated in the hippocampus of aged mice. Results: Red ginseng treatment suppressed the production of age-processed inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor-${\alpha}$, and interleukin-$1{\beta}$ expressions. Moreover, it was observed that red ginseng had an antioxidative effect on aged mice. The suppressed glutathione level in aged mice was restored with red ginseng treatment. The antioxidative-related enzymes Nrf2 and HO-1 were increased with red ginseng treatment. Conclusion: The results revealed that when red ginseng is administered over long periods, age-related decline of learning and memory is ameliorated through anti-inflammatory activity.

• Molecules and Cells
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• v.45 no.12
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• pp.950-962
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• 2022

Aging is a major risk factor for common neurodegenerative diseases. Although multiple molecular, cellular, structural, and functional changes occur in the brain during aging, the involvement of caveolin-2 (Cav-2) in brain ageing remains unknown. We investigated Cav-2 expression in brains of aged mice and its effects on endothelial cells. The human umbilical vein endothelial cells (HUVECs) showed decreased THP-1 adhesion and infiltration when treated with Cav-2 siRNA compared to control siRNA. In contrast, Cav-2 overexpression increased THP-1 adhesion and infiltration in HUVECs. Increased expression of Cav-2 and iba-1 was observed in brains of old mice. Moreover, there were fewer iba-1-positive cells in the brains of aged Cav-2 knockout (KO) mice than of wild-type aged mice. The levels of several chemokines were higher in brains of aged wild-type mice than in young wild-type mice; moreover, chemokine levels were significantly lower in brains of young mice as well as aged Cav-2 KO mice than in their wild-type counterparts. Expression of PECAM1 and VE-cadherin proteins increased in brains of old wild-type mice but was barely detected in brains of young wild-type and Cav-2 KO mice. Collectively, our results suggest that Cav-2 expression increases in the endothelial cells of aged brain, and promotes leukocyte infiltration and age-associated neuroinflammation.

• Journal of Ginseng Research
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• v.42 no.4
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• pp.447-454
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• 2018

Background: Korean ginseng has been widely evaluated to treat human diseases; however, most studies on Korean ginseng have focused on its root. In this study, polysaccharides [acidic-polysaccharide-linked glycopeptide (APGP) extracted with 90% ethanol and hot water] were prepared from Korean ginseng berries, and their effect on immunosenescence was explored. Methods: The effect of APGP on thymic involution was evaluated by measuring the size of thymi dissected from aged mice. The effect of APGP on populations of immune cells, including natural killer (NK) cells, dendritic cells, age-correlated CD11c-positive B cells, and several subtypes of T cells [CD4-positive, CD8-positive, and regulatory (Treg) T cells] in the thymi and spleens of aged mice was analyzed by fluorescence-activated cell sorting analysis. Serum levels of interleukin (IL)-2 and IL-6 were evaluated by enzyme-linked immunosorbent assay analysis. Profiles of APGP components were evaluated by high-performance liquid chromatography (HPLC) analysis. Results: APGP suppressed thymic involution by increasing the weight and areas of thymi in aged mice. APGP increased the population of NK cells, but showed no effect on the population of dendritic cells in the thymi and spleens of aged mice. APGP decreased the population of age-correlated CD11c-positive B cells in the spleens of aged mice. APGP showed no effect on the populations of CD4- and CD8-positive T cells in the thymi of aged mice, whereas it increased the population of Treg cells in the spleens of aged mice. APGP further decreased the reduced serum levels of IL-2 in aged mice, but serum levels of IL-6 were not statistically changed by APGP in aged mice. Finally, HPLC analysis showed that APGP had one major peak at 15 min (a main type of polysaccharide) and a long tail up to 35 min (a mixture of a variety of types of polysaccharides). Conclusion: These results suggested that APGP exerted an anti-immunosenescent effect by suppressing thymic involution and modulating several types of immune cells.

• Korean Journal of Plant Resources
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• v.23 no.6
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• pp.498-502
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• 2010

The immune system may play an important role in aging and the changes in the immune status are associated with treatment of various immunomodulators. This study examined the effects of $\beta$-glucans isolated from mushroom fungi, Coriolus versicolor on macrophages functions in young (8-weeks-old) and aged (82-weeks-old) male C57BL/6 mice. When peritoneal macrophages were treated with various concentrations of $\beta$-glucan ($1-100\;{\mu}g/ml$) for 24 hrs, tumoricidal activity, NO production and phagocytic activity were significantly increased in the young mice, whereas there are no effects in the aged mice. These results suggest that $\beta$-glucans has differential effects on the macrophage functions in young and aged mice and age nutrition might need to be considered to select proper immunomodulator. In addition, $\beta$-glucan could be used clinically for the treatment of diseases such as cancer therapy in the young.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.3
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• pp.269-274
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• 2015

Chronic inflammation has been proposed as one of the main molecular mechanisms of aging and age-related diseases. Although evidence in humans is limited, short-term calorie restriction (CR) has been shown to have anti-inflammatory effects in aged experimental animals. We reported on the long-term treatment of daumone, a synthetic pheromone secreted by Caenorhabditis elegans in an energy deficient environment, extends the life-span and attenuates liver injury in aged mice. The present study examined whether late onset short-term treatment of daumone exerts anti-inflammatory effects in the livers of aged mice. Daumone was administered orally at doses of 2 or 20 mg/kg/day for 5 weeks to 24-month-old male C57BL/6J mice. Increased liver macrophage infiltration and gene expression of proinflammatory cytokines in aged mice were significantly attenuated by daumone treatment, suggesting that short-term oral administration of daumone may have hepatoprotective effects. Daumone also dose-dependently suppressed tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$ )-induced nuclear factor-${\kappa}B$ (NF-${\kappa}B$) phosphorylation in HepG2 cells. The present data demonstrated that short-term treatment of daumone has anti-inflammatory effects in aged mouse livers possibly through suppression of NF-${\kappa}B$ signaling and suggest that daumone may become a lead compound targeting aging and age-associated diseases.

• Journal of Microbiology and Biotechnology
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• v.27 no.12
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• pp.2094-2103
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• 2017

Aging is associated with distinct changes in immune cells and a decline in immune function, leading to increased susceptibility to infection and reduced responses to vaccination. Certain strains of lactic acid bacteria exert beneficial effects on the immune system. Previously, we reported that Weissella cibaria JW15 isolated from kimchi possesses immune stimulatory activity in vitro. In the present study, we further investigated whether oral administration of JW15 improves immune function in aged mice. Eighteen-month-old female mice were administered JW15 daily at low (JW15-L; $1{\times}10^8CFU/mouse$) or high dosage (JW15-H; $1{\times}10^9CFU/mouse$), or with Lactobacillus rhamnosus GG (LGG) using oral gavage. Two-month-old female mice were included as healthy young mice. After 4 weeks, the mice were euthanized and immune profiles were analyzed using whole blood and spleen. In complete blood count analysis, the numbers of white and red blood cells were significantly increased in the JW15-L group compared with those in the old mouse (OM) control group. In addition, administration of either JW15 of LGG resulted in higher numbers of splenocytes in comparison with the OM group. Furthermore, proliferative potentials were higher in all probiotic groups than OM. Cytokines such as IFN-${\gamma}$ and IL-6 were secreted at higher levels in splenocytes isolated from JW15-fed mice than in OM control mice. Similarly, mRNA expression of various cytokines was altered in the JW15 groups. Collectively, these results suggest that JW15 supplementation induces immunomodulatory effects in aged mice and indicate JW15 as a potential probiotic strain to improve immune function in aged animals.

• Journal of Veterinary Clinics
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• v.14 no.1
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• pp.30-36
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• 1997

It is well estabilished that IL-4, IL-6, $IFN{\gamma}$ and immunoglobulin production are increased as a consequence of advancing age, and endothelial cells increase their expression of pro-inflammatory surface protein, such as ICAM-1, as a consequence of contact with inflammatory cytokines, including IL-1, $TNF{\alpha} or INF{\gamma}.$ This study was designed to define the relationship between age-associated changes of cytokines and immunoglobulin production and ICAM-1 expression in aged mice. Serum from aged mice had elevated IL-6 and immunoglobulin levels compared to mature adult controls and activation-induced production of $IFN{\gamma}$ of splenocytes from aged mice were increased above normal adult level. By immunoperoxidase staining, ICAM-1 in hearts of normal adult mice was expressed occasionally at low levels, but in aged mice, the number of positive blood vessels for ICAM-1 were increased and also the immunoreactivity were stronger. Therefore, these finding indicate that the altered expression of ICAM-1 eith normal aging may actually be correlative to these age-related changes in cytokiness and immunoglobulin production.