The CC chemokine receptor 5 (CCR5) is a G protein-coupled receptor that regulates chemotaxis and effector functions of immune cells. It also serves as the major co-receptor for the entry of human immunodeficiency virus (HIV). Recently, CCR5 inhibitors have been developed and used for the treatment or prevention of HIV infections. Additionally, it has been identified that CCR5 controls bone homeostasis by regulating osteoclastogenesis and the communication between osteoblasts and osteoclasts. However, the effects of CCR5 inhibition on bone tissue in elderly patients are unknown. This study aimed to examine the bone phenotype of aged CCR5 knockout (KO) mice. Femoral and tibial bones were isolated from 12-month and 18-month old wild-type (WT) and CCR5 KO mice, and microcomputed tomography and histology analyses were performed. Twelve-month-old CCR5 KO mice exhibited a decreased trabecular bone mass and cortical bone thickness in both femoral and tibial bones compared with agematched WT mice. Eighteen-month-old mice also showed a decreased trabecular bone mass in femurs compared with control WT mice, but not in tibial bones. Unlike in 12-month-old mice, the cortical margin of femurs and tibias in 18-month-old mice were rough, likely because they were aggravated by the deficiency of CCR5. Overall, our data suggest that the deficiency of CCR5 with aging can cause severe bone loss. When CCR5 inhibitors or CCR5 inactivating technologies are used in elderly patients, a preventive strategy for bone loss should be considered.

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.

Gonadotropin releasing hormone (GnRH) centrally plays a role in control of the hypothalamicpituitary- 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.

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.

Aging is a growing concern of age-associated aberrations in immune system or immunosenescence. Probiotics contain not only these general health advantages but also other the regulation of host immune function. Among those probiotics, the immunomodulatory effects of Weissella cibaria JW15 (JW15) strain has already been reported in our previous studies. The objective of this study is to assess improved immunity of JW15 strain in aged mice. In experiment, mice were divided into five groups. Twenty eight–month-old C57BL/6J mice were given a daily dose of 1×109 CFU/ mouse (JW15-H group) and 1×108 CFU/mouse (JW15- L group) of viable JW15. The young mice group (YM) and old mice group (OM) were given PBS. After four weeks of administration, mice were euthanized. Mice blood was collected and analyzed for complete blood cell count (CBC). Then, mice spleen was weighed and analyzed for splenocyte. Finally, we measured the concentration of cytokine secreted by splenocyte and serum. The results showed splenocyte proliferation of JW15-L group by the addition of LPS and concanavalin A was significantly higher than that of OM group. Splenocyte TNF-α production was significantly increased by JW15 intake. In particular, splenocyte TNF-α production of JW15-L group by the addition of LPS (5 μg/mL) was significantly greater than that other group. Splenocyte IL-6 production was also the highest in JW15-L group. Serum IFN-γ production of JW15-L (59.13 ± 16.13 pg/mL) group was significantly higher than that of OM group (39.71 ± 2.51 pg/ mL). When compared to the OM group (32.22 ± 1.92 pg/mL), Serum TNF-α production was significantly increased by JW15-H intake (69.01± 26.51 pg/mL). In conclusion, JW15 enhanced immunomodulative effects. In particular, the effect was significantly excellent in JW15-L group (1 × 108 CFU/mouse). Therefore, Weissella cibaria JW15 would be suitable for consideration as a helpful functional food for companion animals and humans.

This study was conducted for evaluation of the alleviating effects of Phellodendrin cortex water extract (PCWE) on skin inflammation in hairless mice. Skin inflammation was induced by UVB irradiation and application of squalene monohydroperoxide (Sq-OOH) to the back skin of hairless mice for six weeks. At the same time, saline (C), jojoba oil (VC), PCWE (E), and 0.01% retinoic acid diluted with polyethylene glycol (PC) were applied topically twice per day, six days per week for a period of six weeks. The skin erythema index of the E group was lower than that of the C group. Epidermis and dermis of the C group were remarkably thickened, compared to the PC or E group. In the C group, infiltration of many inflammatory cells, including neutrophils and lymphocytes, was observed in dermis, and a large number of mast cells were observed in dermis and hypodermis; the degree of degranulation was remarkable. However, these phenomena were alleviated in the PC and E2 groups. The E group showed a lower activity in skin xanthine oxidase but a higher activity in skin superoxide dismutase, compared to the C group (P<0.05). The VC, PC, and E groups also showed a high activity of skin catalase by 25.3%, 58%, and 42%, respectively, compared to the C group. Taken together, these results indicate that PCWE could have an alleviating efficacy on skin inflammation induced by UVB irradiation and application of Sq-OOH in hairless mice.

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 β-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 β-glucan (1-100 μg/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 β-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, β-glucan could be used clinically for the treatment of diseases such as cancer therapy in the young.