Preimplantation embryonic production in vitro is important in human assisted reproductive technology and animal embryo engineering. Icariin (ICA) is one type of flavonoids and a main component isolated from the stem leaf of Epimedium brevicornum. Flavonoids, which are among the best well-studied natural antioxidants, have been demonstrated to be active in clearing reactive oxygen species (ROS). The purpose of this study was to investigate the effects of ICA treatment during porcine oocyte in vitro aging and their in vitro developmental competency after parthenogenetic activation (PA). Porcine oocytes were matured in vitro for 44 h (control) and for an additional 24 h in the presence of 0, 5 μM ICA (aging, ICA-5), respectively. This study investigated the effects of ICA on nuclear maturation, ROS level, apoptosis index, and the developmental capacity of aged porcine oocytes. Oocyte survival was not different in aging group compared to control or ICA-5 group. The increased ROS level during in vitro aging was prevented in ICA-5 group, while GSH level was not decreased. The decrease of normal spindle formation during in vitro aging was prevented in ICA-5 group. After PA, although the cleavage rate was not different among treatment groups, the blastocyst formation was significantly higher in control and ICA-5 group than in aging group. However, there was significantly difference in the apoptotic index of the ICA-5 group, while it was no difference in the total cell number of the ICA-5 group. (p<0.05). Therefore, this result demonstrated that the ICA is an effective agent to prevent the deterioration during in vitro aging of porcine oocytes.

The citrus flavonoid hesperetin has various pharmacological actions, including antioxidant, anti-inflammatory, and anticancer activities. The purpose of this study is to confirm whether the treatment of hesperetin can protect the oocyte from in vitro aging. Porcine oocytes were matured in vitro for 44 h (control) and for an additional 24 h in the presence of 0, 1, 10, 100, and 250 μM hesperetin (aging, H-1, H-10, H-100 and H-250, respectively). This study investigated the effect of different concentration of hesperetin on maturation, and reactive oxygen species (ROS) level, apoptosis index, and the developmental capacity of aging porcine oocytes. In the results, the percentage of cleaved oocytes that reached to the blastocyst stage of H-100 group (37.9 ± 1.1%) was similar to control (38.1 ± 0.8%), and also significantly higher than other aging groups (23.2 ± 0.8%; H-1, 19.7 ± 1.3%; H-10, 26.7 ± 0.6%; and H-250, 18.4 ± 1.6%.)(p<0.05). The H-100 group was significantly decreased ROS activity, and increased the level of glutathione (GSH) and expression of the antioxidant genes (PRDX5, NFE2L, SOD1 and SOD2) compared to the aging group. The H-100 groups prevented aberrant spindle organization and chromosomal misalignment, blocked the decrease in the level of phosphorylated-p44/42 mitogen-activated protein kinase (MAPK), and increased the mRNA expression of cytoplasmic maturation factor genes (GDF9, CCNB1, BMP15 and MOS). Also, it was confirmed that the H-100 group expressed higher level of estrogen receptor than the aging group. Therefore, this result indicated that hesperetin is an effective agent to protect from the oxidative stress during in vitro aging of porcine oocytes.

Allicin (AL) is one of the biologically active substance in garlic. Many researchers found that AL exhibits strong antioxidant activity and considered to represent anti-aging effect in vitro. The objective of this study was to investigate the effects of allicin treatment during porcine oocyte aging and their in vitro development. The oocyte was maturated in vitro for 44 h (control) without AL or 44+24 h IVM (24 h aging) with 0, 0.1, 1, 10 and 100 μM allicin (0 AL, 0.1 AL, 1 AL, 10 AL and 100 AL). The 1 AL treated group was significantly increased on maturation rate compared to the 0 AL treated groups, but the other treated groups were not different compared to the 0 AL treated group (p < 0.05). The 1 AL treated group was significantly increased on normal spindle formation and chromosome alignment compared to 0 AL treated group. We checked the effect of AL on parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) embryo development using aged oocytes. PA embryos in 1 AL treated group significantly increased the cleavage and blastocyst formation rate compared to control (p < 0.05). However, SCNT embryos of 1 AL treated group were no significant differences in embryo development. In PA embryo quality, the total cell number was significantly higher in 0.1 AL, 1 AL and 10 AL treated groups than control and DNA fragmentation rate in 1 AL was the lowest among all groups. SCNT embryo in 1 AL significantly increased total cell number and decreased DNA fragmentation. The AL treatment on aged oocyte enhanced PA and SCNT embryo developmental capacity. Therefore, AL may be helpful for assisted reproductive technology applying aged oocytes.

In mammal, unfertilized oocytes remain in the oviduct or under in vitro culture, which is called "oocyte aging". This asynchrony negatively affects fertilization in pre- and post-implantation embryo development. Caffeine a phos-phodiesterase inhibitor is known to rescue oocyte aging in several species. The objective of this study is to determine the cytoskeleton distribution in aged oocytes and the embryo developmental ability of aged oocytes in the present or absence of caffeine during maturation. Caffeine treatment increased the incidence of normal spindle assembly of aged oocytes (treatment, 67.57±4.11% aging, 44.61±6.4%) and no significant differences compared to control group. Fluorescence values were compared using ROS (Reactive oxidation species) stain. Fluorescence values appear of con-trol group intensity rate (51.53.±3.80), aging group (68.10±5.54) and treatment of caffeine (45.04±2.98). Aged oocytes that were derived from addition of caffeine to the IVM (in vitro maturation) medium had significantly increased 2-cell that developed to the blastocyst stage compared to the aging group. Blastocysts, derived from caffeine treatment group, significantly increased the total cell number compare aging (90.44±10.18 VS 67.88±7.72). Apoptotic fragments of genomic DNA were measured in individual embryo using TUNEL assay. Blastocyst derived from caffeine treatment group decreased significantly the apoptotic index compared to blastocyst derived from aging group. In conclusion, we inferred that the caffeine treatment during oocyte aging can improve the developmental rate and quality in bovine embryos developing in vitro

The elderly suffer from an impaired immune function being obvious in a higher susceptibility to infections. Although the inflammatory cells are the major immunomodulatory cells, fibroblasts also secrete a variety of inflammatory cytokines and chemokines. Therefore periodontal tissue aging might playa role in development and progress of periodontitis. In this study, we investigated the effect of in vitro periodontal ligament cellular aging on the inflammatory cytokines, chemokines, and matrix metalloprotease(MMP)-2 expression induced by lipopolysaccharide(LPS) treatment. Three different cell populations were used; passages 4-5, 14-15, and 24-25 (at passage 27, more than 90% cells were replicative senescent). LPS increased the expression of interleukin(IL)-1β, IL-6, and tumor necrosis factor-α, IL-8, RANTES, and MMP-2. However, the order of induction folds were passages 14-15 > 4-5 > 24-25. While the expression level of Toll-like receptor(TLR) 4 decreased according to the increase in passage number, the level of TLR2 was highest at passages 14-15 and then decreased at passages 24-25. While the spontaneous expression of IL-8 decreased according to the increase in passage number, that of RANTES and proMMP-2 increased according to the increase in passage number. These results suggest that the aging of periodontal ligament fibroblasts differentially affect the role as immunomodulatory cells in response to periodontopathic bacteria and therefore might be another risk factor of periodontitis progression.

Background : Rehmannia glutinosa root (R. root) has been used as traditional medicine, and is important resource for natural medicines and functional foods. In Korea, various varieties of Rehmannia glutinosa have been developed, and these have various characteristics depending on the variety. Therefore, this study was performed to compare the antioxidant and anti-aging activities of R. root extracts with various varieties (Jihwang 1ho, Goryeo, Daegyeong, Gogang, Togang, Dagang, Wongang, Yeongang, Hwanggang, and Segang). Methods and Results : R. roots were extracted by reflux extraction method with 70% ethanol-water solution at 40℃, and their extracts were used for total phenolic contents, antioxidant and anti-aging activities. The total phenolic content, ABTS and DPPH radical scavenging activities were highest in Yeongang, and these values were 11.50 ㎎/g, 9.37 ㎍/g TEAC, and 7.13 ㎍/g TEAC, respectively. However, the anti-aging activity on human dermal fibroblast cells treated with doxorubicin was higher in Segang, Dagang, and Goryeo than other varieties, and their senescence ratios were measured at 50.66, 57.16, and 61.36% when treated at 200 ㎍/㎖, respectively. The total phenolic content and DPPH radical scavenging activity were negatively correlated with anti-aging activity. Conclusion : These results indicated that the antioxidant and anti-aging activities of R. root extracts were differed according to variety of Rehmannia glutinosa, and positive correlation between anti-oxidant activities and anti-aging activity was not shown.

본 연구에서는 하고초(Prunella vulgaris) 추출물의 항노화 효과에 대하여 알아보고자 하였다. 시험 결과, 하고초 추출물의 콜라겐 생성 촉진 효과는 250 g/mL 농도에서 75.7 %, MMP-1 생성 억제 효과는 200 g/mL 농도에서 90.2 %로 나타났고, 엘라스테이즈 활성 억제 효과는 2.0 % 농도에서 43.7 %로 나타났다. 또한, 하고초 추출물의 항산화 효과 측정을 위한 자유 라티칼 소거 효과 시험 결과 2.0 % 농도에서 76.9 %의 소거 효과를 나타내었으며, 과산화수소에 의한 세포사멸 억제 효과는 2.0 % 농도에서 49.9 %로 나타났다. 하고초 추출물의 피부에 대한 영향을 알아보기 위하여 22명(연령분포 : 34세 ∼ 48세)의 지원자를 대상으로 실시한 이중 맹검 임상 연구 결과, 하고초 추출물 4.0 % 함유 제형을 12주간 도포하였을 때, 대조군과 비교하여 전문가의 육안평가, 모사판을 제작하여 주름의 정도를 분석한 결과 주름 개선에 현저한 효과가 있음을 확인하였다(p < 0.05). 이러한 결과를 통해 하고초 추출물은 피부의 탄력과 주름을 효과적으로 개선함으로써, 천연 항노화 소재로 유용하게 이용될 수 있을 것으로 기대된다.