미선나무(Abeliophyllum distichum Nakai)는 오직 한반도 일부 지역에 제한적으로 자생하는 특산식물이며, 지구범위 인 IUCN Red List Endangered로 평가·등재되어 있는 이화주성(distyly, 二花柱性) 특성을 가진 희귀식물이다. 본 연구는 미선나무 자생 개체군 8개 지역과 천연기념물 개체군 6개 지역 등 총 14개 지역의 정량조사 결과를 바탕으로 미선나무 이화주성에 따른 개화 특성과 개체군 크기를 비교·분석함으로써 향후 미선나무 개체군에 적합한 현지 내 보전관리방안 정립을 목적으로 수행되었다. 개체군별 출현 개체는 전수조사하였으며, 각 개체의 화기 형태로 장주화·단 주화 개체를 식별해 개화 개체를 현장에서 조사·기록 후 분석하였다. 미선나무 전체 출현 개체 수는 총 13,130개체(개화 7,003개체, 미성숙 6,127개체)로 각 개체군의 장주화·단주화 개화 개체 수 적정 균형 상태가 유의적(P<0.05)이지 않고, 불균형한 상태를 나타냈다. 특히, 영동 개체군은 타 개체군 대비 매우 불균형적 형태를 나타내 타 개체군과 비교했을 때 유전다양성이 낮고, 근친교배 가능성이 클 것으로 추정된다. 관리 단위별 평균 개화·결실률은 각각 자생 개체군 39.0%, 8.5%, 천연기념물 개체군 89.2%, 55.3%로 천연기념물 개체군이 자생 개체군 대비 매우 높게 나타났다. 이는 상층 수관 울폐 차이에 따른 임내 광유입 변화와 관계된 생식 생장 크기 차이로 사료된다. 한반도 내 미선나무 전체 점유면적은 23,224.5㎡이었고, 천연기념물 개체군이 자생 개체군 대비 개체군 점유 면적은 작으나 밀도는 상대적으로 높게 나타났다. 일정 면적에 대한 보호시설 설치로 개체군 확산에 제약이 있는 천연기념물 개체군 내 주기적인 관리가 천연기념물 개체군 밀도를 높인 주된 요인일 것으로 추정된다. 미선나무 보전을 위해서는 현재 인위적·유전적 교란이 의심되는 천연기념물 집단을 해제하고, 새로운 보호구역을 신규로 지정함으로써 보전 우선 개체군의 확대가 필요하다. 천연기념물 개체군의 경우 문화재청과 지자체에서 자생지 관리가 이뤄지고 있으나 자생 개체군의 경우 관리 주체가 전무한 실정으로 미선나무 전반적 관리에 대한 제도적 개선이 우선적으로 이뤄져야 할 것이다.

Abeliophyllum distichum Nakai is a deciduous shrub of a flowering plant in Oleaceae. It is an important plant resource and consists of only one species in the entire world. A. distichum Nakai is well known an edible, medicinal herb in its habitat districts, but the toxicological evaluation for the safe use of its extract is still insufficient. The study characterized the toxicity of an Abeliophyllum distichum Nakai ethanol extract in Sprague-Dawley (SD) rats and determined the safe dosage levels in a 13 weeks toxicity study. Abeliophyllum distichum Nakai ethanol extract was orally administered once daily for 2 weeks at 0, 500, 1,000 and 2,000 mg/kg/day to male and female SD rats. while recording the clinical signs of toxicity, body weight, food intake/consumption, eye test and urine analysis. Only the total protein frequency in the urine of male SD rats (p<0.05), the right ovary of the 500 mg/kg group (p<0.01) and the right adrenal gland of the 1,000 mg/kg group (p<0.05) in the female rats showed statistically significant changes. But no toxic effects were noted from repeated-dose administration of the Abeliophyllum distichum Nakai ethanol extract in the SD rats during the observation period. The post-mortem examinations showed no test substance-mediated changes. The hematological analysis and clinical blood chemistry data demonstrated no toxic effects from repeated-dose administration of Abeliophyllum distichum Nakai ethanol extract in the SD rats during the observation period. Based on these results, this data suggests that a dose of 1,000 mg/kg/day is a highest treatment to administer when conducting a further 13 weeks toxicity study.

본 연구는 새롭게 확인된 경기도 여주시 미선나무 자생지를 대상으로 식생현황 및 분포규모와 신규 자생지의 의미를 분석하고 향후의 보전 및 관리에 필요한 기초자료를 제공하기 위해 수행되었다. 미선나무는 세계에서 우리나라에만 분포하는 특산식물로 1속 1종만이 존재한다. 그동안 보고된 미선나무 자생지는 충청지역을 중심으로 하는 중남부 지역에 편중되어 있었으나 여주시 자생지의 발견으로 분포역이 중부지역까지 확장되었다. 대상지는 37˚20~21´N, 137˚43´E, 해발 99~120m에 위치하고 있으며, 하천을 끼고 있는 산자락 전석지대의 지형적 특징을 보이고 있다. 자생지의 규모는 약 530㎡로 좁은 면적이었으며, 자생지 내에 미선나무는 약 1,200여 개체가 분포하고 있다. 출현하고 있는 개체들은 수고 0.5m 이하의 어린 개체가 대부분이었고, 1.0m 이상의 성숙한 개체는 약 300여 개체로 확인되었다. 여주시의 미선나무는 뿌리나 줄기를 이용한 영양번식에 의해 개체군이 유지 및 확산되고 있다. 자생지의 식생은 교목층에서는 리기다소나무와 갈참나무가 우점하고, 아교목층은 생강나무, 갈참나무, 신나무 등이 출현하고 있다. 관목층은 미선나무가 우점하는 가운데 국수나무, 회잎나무, 쥐똥나무 등이 출현하였으며, 초본층은 낮은 식피율을 보였다.

In this study, we evaluated the whitening activity of prethanol A and water extracts from Abeliophyllum distichum Nakai. The extracts were prepared using 0, 50, 70, and 100% prethanol A at 121℃, 1.2 atm for 15 minutes. To confirm effective extraction, the acteoside content of each extract was analyzed with the HPLC-PDA method. The antioxidant activity was evaluated using DPPH and ABTS scavenging activity assays, and the whitening activity was evaluated based on inhibitory activities on the protein and mRNA expression of tyrosinase, tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 2 (TRP-2), and microphthalmia-associated transcription factor (MITF) in B16 F10 cells. Each extract showed strong antioxidant and whitening activity. IC50 values of antioxidant activity from each extract were in order of 100%, 70%, 50%, and 0%. In addition, whitening activity inhibited the protein and mRNA expression of melanin synthesis factor, following the same pattern as antioxidant activity. In conclusion, water and prethanol A extracts of A. distichum showed effective antioxidant and whitening activity and are thus considered to be valuable materials for whitening cosmetics. The results of this study will also provide basic data for the safe and efficient production of A. distichum as a cosmetic material.

In this study, we evaluated the antioxidant activity and protective effects against oxidative DNA damage of the ethyl acetate fraction from the callus of Abeliophyllum distichum Nakai (ECA). Callus of A. distichum was induced on MS medium containing NAA (1 ㎎/L) and 2,4-D (1 ㎎/L), and a sufficient amount was obtained for the extraction by subculture. Acteoside was analyzed and quantified (0.39 ㎎/g callus) from ECA using the high-performance liquid chromatographyphotodiode array detector method. ECA showed very high antioxidative activity as revealed by DPPH and ABTS scavenging assays. The IC50 values were 12.4 and 6.8 ㎍/㎖, respectively. ECA showed protective effects against oxidative DNA damage evaluated by using ψX-174 RF I plasmid DNA. It also inhibited DNA damage by suppressing the oxidative stress-induced protein and mRNA levels of γ-H2AX and p53 in NIH/3T3 cells. In conclusion, ECA protects against oxidative DNA damage through its powerful antioxidant activity.

In this study, we evaluated the antioxidant activity and anti-inflammatory effects of Abeliophyllum distichum (A. distichum) leaves that were prepared via air-drying. Fresh and air-dried A. distichum leaves were examined via 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging assay and measurements of the reducing power. The suppression effects on inflammation of the leaves were analyzed by a western blot and RT-PCR on LPS-induced RAW 264.7 cells. As a result, the antioxidant activity of the fresh leaves was found to be more effective than that of the air-dried leaves. Also, the fresh leaves were more effective in suppressing the protein and mRNA levels of iNOS and COX-2 than the air-dried leaves, thereby indicating the better anti-inflammatory effects. In addition, the contents of phenolic compounds and acteoside were analyzed by high-performance liquid chromatography (HPLC). The results showed that the acteoside content decreased with the use of the air-drying method, while there was no change in the content of phenolic compounds. Therefore, this study indicated that fresh A. distichum leaves potential antioxidant and suppression activities of various factors that are involved in the production of NO, which were found to be better than those of air-dried A. distichum leaves. These biological activities were also found to be independent of the content of phonolic compounds and were assumed to be directly or indirectly related to the content of acteoside.

Abeliophyllum distichum Nakai (A. distichum) has been reported to exert the inhibitory effect on angiotensin converting enzyme and aldose reductase. Recently, our group found that branch extracts of A. distichum (EAFAD-B) induce apoptosis through ATF3 activation in human colon cancer cells. However, anti-cancer reagents exert their activity through the regulation of various molecular targets. Therefore, the elucidation of potential mechanisms of EAFAD-B for anti-cancer activity may be necessary. To elucidate the potential mechanism of EAFAD-B for anti-cancer activity, we evaluated the regulation of cyclin D1 in human colon cancer cells. EAFAD-B decreased cellular accumulation of cyclin D1 protein. However, cyclin D1 mRNA was not changed by EAFAD-B. Inhibition of proteasomal degradation by MG132 attenuated EAFAD-B-mediated cyclin D1 downregulation and the half-life of cyclin D1 was decreased in the cells treated with EAFAD-B. In addition, EAFAD-B induced cyclin D1 phosphorylation at threonine-286 and the point mutation of threonine-286 to alanine attenuated EAFAD-B-mediated cyclin D1 proteasomal degradation. Inhibitions of both ERK1/2 by PD98059 and NF-κB by a selective inhibitor, BAY 11-7082 suppressed cyclin D1 downregulation by EAFAD-B. From these results, we suggest that EAFAD-B-mediated cyclin D1 downregulation may result from proteasomal degradation through its threonine-286 phosphorylation via ERK1/2-dependent NF-κB activation. The current study provides new mechanistic link between EAFAD-B and anti-cancer activity in human colon cancer cells.

Abeliophyllum distichum is a monotypic taxon of Oleaceae and endemic to Korea. A comprehensive study on embryogeny and fruit and seed coat ontogeny in Abeliophyllum was carried out via microtome and light microscopy. The fertilization occurs during mid– to late April and embryo matures by early July. The embryo development follows the general fashion from globular embryo – transition embryo – heart shaped embryo – torpedo embryo – walking-stick embryo to mature embryo. The pericarp clearly differentiates into three histological zones: exocarp, mesocarp, and endocarp. The young seed comprises 10-12 cells thick seed coat and the mature seed coat comprises an exotesta, 6-8 mesotesta and an endotesta. Any crystals, phenolic-like compounds, idioblasts, and the sclereids are not found in pericarp as well as seed coat. An overall development confirms Solanade type of embryogenesis in Abeliophyllum. The endocarp becomes more prominent in mature fruit

Skin anti-wrinkle activities of the stems and leaves of Abeliophyllum distichum Nakai were evaluated by theextracts obtained from various extraction processes such as using hot water at 100oC, 70% ethanol at 85oC, and 70% etha-nol with ultrasonication at 60oC The ultrasonicated extract showed 95.62% of the highest cell viability in addition of0.3㎎/㎖ of the extracts into the normal human fibroblast cell, CCD-986sk. For antioxidant activities, the extracts usingultrasonicated extract showed the highest DPPH free radical scavenging as 80.27%, followed by 75.88% and 62.44% for theextracts using ethanol extract and water extract. The ultrasonicated extract also showed the highest elastase inhibition activ-ity as 25.32%, compared to ethanol extract and water extract based method at 22.01% and 12.88%, respectively. MMP-1production was most effectively decreased down to 2908.1pg/㎖ with ultrasonicated extract while 6640.8pg/㎖ with waterextract and 3609.3pg/㎖ with ethanol extract, in addition of 0.3㎎/㎖. Collagen production was increased up to 154.7ng/㎖in addition of ultrasonicated extract, and followed by 121.4ng/㎖ and 31.2ng/㎖ for ethanol extract and water extract,respectively. These results indicate that the ethanol extract should have skin anti-wrinkling activities and can be improvedby the ultrasonication process that high energy input elute more amounts of bioactive substances eluting more amounts ofbioactive substances from the high energy input of ultrasonication.

This research evaluate antioxidant and skin-whitening effect of Abeliophyllum distichum Nakai by extraction processes. First, antioxidant effects were follows: EE (70% ethanol extract) showed higher DPPH scavenging activity of 69.66% than WE (hot water exract) 59.13% at 0.3 ㎎/㎖, also UE's (70% ethanol extract by sonication process) higher than EE. Reducing power was that also EE showed higher than WE, and it was the highest value with UE's because of ultrasonic pretreatment. Next, the whitening effect tyrosinase inhibition activity was measured that EE was 23.88%, WE's was 16.69%, and UE was 23.34%. Ultrasonic pretreatment did not influence to tyrosinase inhibition activity. Cell viability showed low cell toxicity in all groups. UE's inhibited melanin synthesis, 55.1%, that is higher than EE and WE, 52.7% and 39.5%, respectively. As a result, we confirmed that antioxidant activities and skin-whitening effect by extraction process. Also, this results confirmed that the Abeliophyllum distichum Nakai extracts worth as cosmetic materials.

The present study attempts to evaluate antioxidant activities of extracts from Abeliophyllum distichum. Nakai flower. The samples were collected in Jangyyeon-myeon, Goesan-gun, Korea and extracted with either hot-water or ethyl acetate (EtOAC). In DPPH, hydroxyl radical scavenging activity and Fe2+ chelating activity of EtOAC extracts were 93.41%, 98.43%, and 7.38%, while those of hot-water extracts were 86.93%, 41.33% and 47.68% at 200 μg/ml, respectively. In ΦX-174 RF I plasmid DNA cleavage assay, the protective effects of EtOAC and hot-water extracts against oxidative DNA damage were 82% and 17% at 200 μg/ml, respectively. Both extracts showed the protective effect of DNA migration by oxidative stress in intracellular DNA migration assay. Both extracts had no cytotoxity in NIH3T3 cells. Several polyphenolic compounds were identified such as 2-methoxy-benzoic acid, vanillic acid, phytol and pulegone by GC/MS. These results indicated that extracts of Abeliophyllum distichum Nakai flower showed antioxidant activities and protective activities against oxidative DNA damage and showed the possibility to be used as an effective natural antioxidants.

This experiments were carried out to find out the effects of different explant materials, kinds and concentration of plant growth regulators, and total nitrogen and sucrose contents on the in vitro regeneration of Abeliophyllum distichum Nakai. The effects of growth regulators on regeneration from 3 explant sources (leaf, internode and node) were more or less same. Leaf explants produced only callus with 2ip (Isopentenyladenine) and NAA (Naphthaleneacetic acid) treatment and other regulators had no effects. Test with internode explants yielded about same results but callus was obtained with 2,4-D (2,4-Dichlorophenoxyacetic acid). Node explants resulted in shoot regeneration by all regulator treatment except NAA and 2,4-D, but control also showed similar results. Callus formation from internode and node explants was vigorous by 2ip, zeatin, and 2,4-D treatments and high NAA concentration resulted in higher callus formation. In this experiment, various mixed treatment of growth regulators were also employed, using node as explant material. Shoot regeneration was obtained with BA (Benzyl adenine) + NAA treatments but the results were comparable with control. Generally shoot and root regeneration was poor with all combined treatment except 2ip + NAA and 2,4-D + NAA. However, callus was formed readily with all treatments. In this experiment, combined treatments of regulators were applied on the callus derived from singular regulator treatment. The results showed no shoot and root regeneration with any combination of 2,4-D, IAA (Indoleacetic acid) and NAA, but soft milky white callus was formed in all the treatments. No shoot and root regeneration was observed with any combination of 2iP, NAA and IAA, but somewhat hard, light green callus was formed in all the treatments. Callus formation decreased with high kinetin concentration in case of kinetin + NAA treatment. The experiments with total nitrogen content of media showed that low concentrations of 15 and 30mM were effective for the shoot and root regeneration. Sucrose experiment demonstrated shoot regeneration with 1~4% concentration, and root and callus formation with 2~4%. No root and callus formation was observed with 0 and 1% sucrose.