본 연구에서 사용된 국내산과 중국산 댕댕이나무 열매 는 평균 길이가 약 2.2 cm이고, 자흑색의 타원형으로 육안 상 국내산과 중국산 댕댕이나무 열매는 비슷한 외형을 보 인다. 국내산과 중국산 댕댕이나무 열매를 기능성 식품소 재로 활용하기 위하여 polyphenol, amino acid 및 fatty acid 를 분석하고, 항염증 활성을 측정하여 원산지에 따른 차 이를 비교하였다. 총 플라보노이드 함량은 국내산 열수 추 출물과 30% 에탄올 추출물에서 각각 8.58±0.11 mg QE/g 과 11.83±0.25 mg QE/g의 결과를 보였고, 중국산 열수 추 출물과 30% 에탄올 추출물에서 각각 3.49±0.09 mg QE/g 과 4.46±0.10 mg QE/g의 결과를 보여 국내산이 중국산 열수 추출물과 30% 에탄올 추출보다 2배 이상 높았다. Chlorogenic acid, caffeic acid 및 rutin을 분석한 결과, chlorogenic acid 함량이 국내산 열수 추출물과 30% 에탄 올 추출물에서 각각 1.51±0.02 mg/g과 1.94±0.03 mg/g으 로 중국산 열수 추출물과 30% 에탄올 추출물에서 각각 0.23±0.00 mg/g과 0.25±0.01 mg/g을 보인 결과보다 7배 이상 높았으며, caffeic acid와 rutin은 모든 추출물에서 미 량 또는 낮은 함량을 보였다. 아미노산 분석결과, 국내산 열수 추출물과 30% 에탄올 추출물에서 17종의 아미노산 이 확인되었고, 중국산 열수 추출물과 30% 에탄올 추출 물에서 cystine을 제외한 16종의 아미노산이 확인되었으며, 주요 아미노산은 glutamic acid로 확인되었다. 총 아미노 산 함량은 국내산 열수 추출물과 30% 에탄올 추출물, 중 국산 열수 추출물과 30% 에탄올 추출물에서 각각 2.02±0.03 g/100 g과 0.81±0.03 g/100 g, 0.15±0.01 g/100 g 과 0.05±0.00 g/100 g의 결과를 보였다. 지방산 분석결과, 국내산 열수 추출물과 30% 에탄올 추출물 및 중국산 30% 에탄올 추출물에서 caprylic acid (C8:0)를 제외한 13종 지 방산이 확인되었고, 중국산 열수 추출물에서 caprylic acid (C8:0)와 lignoceric acid (C24:0)를 제외한 12종 지방산이 확인되었으며, 모든 추출물에서 주요 지방산은 palmitic acid (C16:0)로 확인되었다. 지방산 조성은 포화지방산이 국내산 및 중국산 열수 추출물과 30% 에탄올 추출물에서 52.02±2.00- 84.17±3.51%로 대부분을 차지하였고, 총 지방산 함량은 국 내산 열수 추출물과 30% 에탄올 추출물, 중국산 열수 추출 물과 30% 에탄올 추출물에서 각각 0.49±0.06 mg/g과 1.12±0.03 mg/g, 0.45±0.03 mg/g과 0.76±0.02 mg/g을 보였 으며, 모든 추출물에서 지방산 함유량이 대체로 낮은 결과를 보였다. 세포독성 측정결과, 국내산 및 중국산 열수 추출물과 30% 에탄올 추출물은 500-2,000 μg/mL 농도에 서 RAW 264.7 대식세포에 대한 독성이 나타내지 않았다. NO 생성억제 활성은 국내산 및 중국산 열수 추출물과 30% 에탄올 추출물의 농도에 따라 유의적으로 NO의 생 성량을 감소시키는 것으로 확인되었고, cytokine 생성억제 활성 또한 TNF-α와 IL-6의 생성량이 유의적인 감소를 보 여 항염증 활성을 나타내는 것으로 확인되었다. 이러한 연 구결과를 통하여 댕댕이나무 열매를 활용한 기능성 식품 소재 연구에 이용될 수 있을 것으로 사료된다.

Lonicera caerulea (Honey berry, HB) has been used in medical treatment in Russia, Japan, China and Korea. It has high level of vitamin C and polyphenolics. Polyphenolics can improve anti-inflammatory effect and prevent cancer, diabetes mellitus type 2. Also, Vitamin C is a representative anti-oxidant. however, it is still unknown what effect it will have on the oxidation stress of the reproductive system. In previous studies, ROS can be produced when it is exposed to heat stress and has negative effect on sperm’s maturation, capacitation, hyperactivation, acrosome reaction and fusion of egg and sperm. Therefore, the purpose of this study is to investigate the antioxidant effects of L. Caerulea on the sperm and mice. At first, it conducted using ICR mouse (n = 20) for 4 weeks. There are four groups of mice (n = 5 per group). Also, L. Caerulea was taken by oral gavage. Group I (control) kept at 23℃-27℃ and administer D.W (0.5 mL/day), Likewise, Group II (HB) kept at room temperature but gave HB (250 mg/kg, 0.5 mL/day), Group III (HB + HS) received heat stress (40℃) using hyperthermia induction chamber and gave HB at same dose. and Group IV (HS) exposed heat stress only. Mainly, we showed degree of gene expression using Western blot in SOD, HSP 70, 17β-HSD and Real-time PCR. It can find correlation between intracellular activity like steroid hormone, apoptosis under ROS and antioxidant activity of L. Caerulea.

Mitochondria is energy generating organelle. It synthesizes ATP, which is the essential energy source of many cellular processes. During producing energy, some redox centres leak electrons to oxygen and it is contributory to the reactive oxygen species. Besides, mitochondria have significant functions in metabolism, calcium homeostasis, and fatty acid oxidation. Also mitochondria has importance to the breakdown of the ovarian follicles and could be factor determining oocyte of quality adversely. Increasing evidence shows that the number of mitochondria affect oocyte of developmental competence and maturation detrimentally during aging. Oocyte is the mitochondria-rich cell and enable the organelle to have competence for fertilization and early embryonic development. Occurrence of blastomere depends on distribution change of mitochondria which present in the egg. Lonicera caerulea treatment inhibited ovarian mitochondrial oxidative damage by suppressing mitochondrial reactive oxygen species (mROS) generation, decreasing apoptosis, controlling disintegration of mitochondrial membrane potential and conserving respiratory chain complex activities. The purpose of this study is to identify if mouse accepting treatment with L. caerulea could counter age-induced sterility and ovarian mitochondrial OS in a model organism of ovarian ageing.

Mammalian oocytes are sensitive to psychological stress at each period of follicular development. Especially, thermal stress interfere with reproductive condition by inducing formation of reactive oxygen species (ROS) and oxidative stress (OS). ROS lead to oocyte apoptosis, weakening oocyte quality and lowering the fertilization rate. As a result, the pregnancy rate is lowered, leading to infertility. Thermal stress also seems to influence zygotes through physiological changes in the maternal environment surrounding them. Loss of developmental competence suggests hyperthermia-induced oxidative stress in embryos. Interest in organic Lonicera caerulea berries has increased in recent years. They are abundant in various health-improving materials. Berries that found from natural products can be as free as possible from the bioactive toxicity of the active ingredient without side effects, and it can be a big advantage because it can work. Mammalian oocytes are arrested at the first meiotic prophase stage and get their meiotic competence to produce offspring during the development of follicle. A series of nuclear and cytoplasmic maturations are involved in this process and these vary in temperature sensitivity. Our study demonstrated that L. caerulea can relieve the negative effects of maternal hyperthermia by reducing ROS level at the developmental stage.

Lonicera Caerulea(Honey berry) has been used in medical treatment in Russia, Japan, China and Korea. It has high level of vitamin C and polyphenolics. Polyphenolics can improve anti-inflammatory effect and prevent cancer, diabetes mellitus type 2. Also, Vitamin C is a representative anti- oxidant. however, It is still unknown what effect it will have on the oxidation stress of the reproductive system. In previous studies, ROS can be produced when it is exposed to heat stress and has negative effect on sperm's maturation, capacitation, hyperactivation, acrosome reaction and fusion of egg and sperm. Therefore, The purpose of this study is to investigate the antioxidant effects of L.caerulea on the sperm and egg cells of mice. At first, it conducted using ICR mouse(n=20) during 4 weeks. There are four groups of mouse(n=5 per group). Also, L.caerulea was taken by oral gavage. Group Ⅰ(control) kept at 23℃~27℃ and administer D.W(0.5ml/day), Likewise, Group Ⅱ(HB) kept at room temperature but gave HB(0.5ml/day), Group Ⅲ (HB+HS) received heat stress (40℃) using hyperthermia induction chamber and gave HB at same dose. and Group Ⅳ(HS) exposed heat stress only. Mainly, we showed degree of gene expression using Western blot in SOD, HSP 70, 17β-HSD and Real time PCR. It can find correlation between intracellular activity like steroid hormone, apoptosis under ROS and antioxidant activity of L.caerulea.

Previous studies have shown that Lonicera caerulea has a chemical protective effect. Phenolic and vitamin C contained in Lonicera caerulea prevent cancer, diabetes and cardiovascular disease, lower blood pressure and delay the aging process. However, the antioxidant mechanism of male reproductive system to heat stress is still unknown. Male reproductive system is very sensitive to heat. When scrotum temperature increase, oxidative stress can occur. Oxidative stress affects sperm motility and spermatogenesis, resulting in infertility. Therefore, we investigated the antioxidant effect of L. Caerulea in male genitalia by inducing oxidative stress by artificially exposing the testicles to heat at 42 ° C. The experiment was performed by dividing the ICR mouse into four groups. Each group is n = 5. Control group (C) and heat stress group (HS) were oral gavage administered D.W. Honey berry group (HB) and Honeyberry / heat stress group (HB + HS) were oral gavage administered honey berry (250mg / kg / day). HS groups (n=5, per n=5) received heat stress by exposing their lower bodies in the water bath at 42℃ for 30 minutes. We confirmed that there was a significant difference in the motility, morphology and the number of sperms using CASA(computer-assisted semen analysis). Lipid peroxidation assay results showed heat causes oxidative stress in serum. This study is conducting to investigate the antioxidant effect of L. Caerulea. Histologically analyzed the testicular form of each group, the activity level of heat shock protein and the level of reactive oxygen species were measured by Western blot and the level of catalase and HSP-90 was examined by RT-PCR analysis. Thus, studies of testicular morphology, sperm kinetics, hormone levels, heat shock protein expression and antioxidant enzymes under heat stress have shown that L. Caerulea ingestion has Anti-oxidant and thermal protective activity on the testis by heat damage.

이상의 연구 결과로 미루어 볼 때, 댕댕이나무 잎과 가지 추추출물은 대장암 세포주 HCT116과 SW480세포의 생육을 억제 하였으나 열매추출물은 억제활성이 나타나지 않았다. 잎과 가지 추출물은 cell migration과 wound healing assay를 통해 비정상적인 세포증식 억제를 확인하였으며, β-catenin과 TCF4 의 단백질 수준을 감소시켜 비정상적인 Wnt 신호전달을 억제를 통해 대장암세포의 생육을 억제하는 것으로 판단된다. 따라서 댕댕이나무 잎과 가지는 항암을 위한 대체보완소재 및 천연 항암제 개발을 위한 소재로 활용이 가능할 것으로 판단된다.

Background : Haskap berries commonly refer to fruits of Lonicera caerulea L., recognized by the Japanese aborigines as the “The elixir of life.”. Due to their recent arrival on the North American market, haskap berries have not yet been positioned among other berries and compared in terms of their phytochemical content. And haskap berries have higher ascorbic acid and anthocyanin content than other berries known for their health-promoting benefits, such as blueberries. However, no study has reported on the antioxidant and anti-cancer activity of Lonicera caerulea stem. The purpose of this study is to present the current research on the chemical content, antioxidant and anti-cancer activities of Lonicera caerulea stem. Methods and Results : The stem of Lonicera caerulea L. ware dried in the shade at room temperature and extracted with 100% methanol. The extract was suspended in deionized water and partitioned sequentially with n-hexane, chloroform, ethyl-acetate and butanol (water saturated BuOH) fractions. Antioxidant activities were measured by determination of antioxidants, DPPH (2,2-diphenyl-1-picrylhydrazyl). Cell viability was determined by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. All cell lines were purchased from the Korean Cell Line Bank (Seoul, Korea). All results were performed with three replications were processed statistically. By DPPH assay, the Lonicera caerulea L. the highest activity was obtained from the ethyl-acetate fraction (IC50=15.46 ㎍/㎖). By MTT assay, the chloroform fraction showed a significant growth inhibiting effect on MCF-7 (Human breast cancer, IC50=225.91 ㎍/㎖), COLO 205 (Human colon cancer, IC50=179.55 ㎍/㎖), but on AGS (Human stomach cancer) and other fractions it did not show effect. Conclusion : We demonstrated that Lonicera caerulea L. stem extract and fractions has antioxidant and antiproliferation activity in vitro. Further studies should identify the active constituents in Lonicera caerulea L stem to evaluate the potential in vitro antioxidant and antiproliferation activities of the extract.

Lonicera caerulea var. edulis is a rare species found in some alpine region of Korea. Genetic variation in L. caerulea var. edulis has been investigated by examining 161 individuals from six natural populations: Mt. Seorak 1, Mt. Seorak 2, Mt. Jeombong, Mt. Bangtae, Mt. Gyebang, Mt. Halla. The mean genetic diversity for all the six populations was 0.25 (S.I.). The highest genetic diversity was found in Mt. Seorak (S.I.=0.3158) and the lowest was in Mt. Gyebang (S.I.=0.1047). Comparatively low level of genetic diversity was observed (Ae=1.25, P= 64.6%, S.I.=0.25), which is a typical pattern for rare tree species. AMOVA showed exceptionally large proportion of genetic variations both for among populations (34.69%) and within populations (65.31%). Excluding Mt. Gyebang, the genetic variation among and within population was 18.71% and 81.29% respectively. The UPGMA dendrogram based on genetic distance is not suitable for geographic relationship. Genetic distance of Mt. Gyebang was most distant from the other populations. Excluding Mt. Gyebang, the genetic identities among the five populations were 0.95 to 0.97, which is very high similarity level of genetic identity. This low level of genetic variations and the lack of site in nature indicates that L. caerulea var. edulis demanded a serious conservation.