RAPD 마커를 이용하여 인삼 품종 및 육성계통의 유전적 다양성 및 유연관계를 분석한 결과는 다음과 같다.
1. 총 130개의 primer 중 polymorphism을 나타내는 70개의primer를 선발하였고, 그 중 재현성이 있으면서 polymorphism이 높은 25개의 primer를 선발하였다. 증폭된 DNA 단편의수는 189개이고, PCR 산물은 100 ~ 2,800 bp 범위로 증폭되었다.
2. 각 primer에 의해 증폭된 DNA 단편의 수는 3개 ~ 17개로 다양하였으며, primer 한 개당 평균 7.6개의 DNA 단편이증폭되었다. OPD19 primer를 이용한 유전분석 결과, 총 5개의 유전양상이 나타났는데, 약 500 ~ 1,300 bp의 증폭산물에서품종 및 계통 간 유전적 다형성을 나타냈다.
3. 선발된 primer별 대립인자는 최소 1.33에서 최대 2.00의 범위였고, 평균 1.709이었다. primer별 유전적 다양성은 OPD15가 가장 높았고, OPF2가 가장 낮은 값을 나타내었다. 본 연구에서 분석에 이용된 25개의 RAPD primer 중에서 D15, D19, B5, A19등은 인삼 품종과 계통에서 비교적 높은 수의 대립단편과 높은 유전적 다양성 값을 나타내는 primer였다.
4. 유사도 계수 0.98을 기준으로 24개의 품종 및 계통을 대상으로 군집분석을 수행한 결과, 미국에서 수집 육성된 G04116과 국내 품종인 천풍, 연풍 그리고 국내 육성 계통인 G04009, G04026, G04069, G04084는 그룹을 형성하지 않았고, 17개의 품종 및 계통은 2그룹으로 분류되었다. I 그룹에는 고풍, 금풍과 12계통(85%), II 그룹에 3계통(15%)이 포함되었다.
The purpose of this study was to examine the effects of vi tamin D3 and 1'etinoic acid(RA) on the human mesenchymal stem ce!ls(MSC) g1'owth and osteogenic differentiations. Cell proliferation, mineralization, cell cycle, expression of cell cycle regu l atOJγ proteins and markers fo1' osteogenic differenatiaiton were determined by MTI assay, mineralization assay, flow cytomet1'Y‘ and Western blot analysis, respectively. Cell viability was dec1'ease by each vitamin D3 and RA added to MSC. it was more decrease by vitamin D3 and RA. Mineralized nodule formation revealed similar expression pattern with positive cont rol group at vitamin D3 and RA mixed add to MSC. At vitamin D3 and RA mixed add to MSC after 7 days of incubation was increase G1 s tage. after 21 days of incubation was inhibit cell cycle prog1'ess by inc1'ease of sub-G1 Treatment vitamin D3 to MSC inhibits p53 and p21, but inc1'ease pRb. RA inhibit p53, but increase p21 and pRb, vitamin D3 plus RA group was same as added RA group. so two vitamin was effect to inhibited cell growth each different mechanism. Expression of BMP-2 protein was prominent in osteogonic supplement treated g1'oup of MSC at 2 weeks cultivation days, but vi tamin D3 treatment decreased BMP-2 expression rather than in (+) control group. BSP protein was notably increased in the OS compa red to positive controls at 2 weeks cultivation, but similar to that of vitamin D3 group t1'eatment group and was least expressed in plus RA mixed group, at 3 weeks, BSP expression was similar to 1'esult of 2 weeks Collectively, these results shows that vitamin D3 and RA have diffe1'ential effects on the MSCs g1'owth and differ entia tion 211
We have examined the effect of NO donor, S-nitl‘ oso-N-acetyl-DL-penicillamine(SNAP) on heme oxygenase-1 (HQ-l) ex pression in human oral immortalized & malignant keratinocytes, and investigated in the control of keratinocyte proliferation evidence tha t HO-1 cou ld be involved in a low dose of NO, NO inhibitor, HOinducer, and HO inhibitor medi ated cytoprotect ion against cytotoxi city induced by a high dose of NO Oral keratinocyte growth inhibitory or anti-proliferative effects were exerted by with SNAP and hemin in a dose- and cul tivation time dependent manner The level of HQ-1 protein was increased in all cell types after exposure hernin dose, and the hemin induced HQ-1 protein achieved at higher maximum level by 12 hrs in all kind of cells , The pretreatment of cells with 0, 2 μ M SNAP reduced 1 mM SNAP-induced death in IHOK and HN4 cells , These cytoprotective effects on high dose of NO induced HQ-1 expresion and cell ular toxicity were blocked by low dose of SNAP, HCB, and ZnPP IX supporting the involvement of HQ-1 in high dose NO induced growth arrest or cell death, But these cytoprotection pattern is different from immortalized and malignant keratinocytes , These results indirectly demonstrate that HQ-1 could be involved in cytoprotection by NO priming against high dose NO induced cytotoxicity in immortalized and maigla nt oral keratinocytes, Thus, HQ-1 might be an important cellular target of NO donor, with clinical implications for the pre vention of inJlammatory di seases and anti-tumor immunity
To study on antioxidant effects in the liver of 40-week-old mouse, the sample were orally pretreated 5mg/kg/day for 5 days with red ginseng saponin components(total saponin, protopanaxadiol saponin, protopanaxatriol saponin, ginsenoside-Rd, ginsenoside-Re, compound-K) for 5 days. The ability of saponin to protect the mouse liver from oxidative damage was examined by determining the activity of superoxide dismutase(SOD), glutathione peroxidase(GPx) and the contents of glutathione, the level of malondialdehyde, The only protopanaxadiol among the ginseng saponin fractions was significantly increased the hepatic SOD activity(p<0.01). The red ginseng saponin induced a slight increase of GPx activity, especially ginsenoside Rd, compound K and protopanaxatriol treatments significantly increased its activity. The content of glutathione was significantly increased by total saponin, protopanaxadiol and ginsenoside Rd(p<0.01), but the oxidized glutathione level was lowered in all the red ginseng saponin. Finally, the level of malondialdehyde was significantly decreased by ginsenoside Rd and protopanaxadiol. In conclusion, protopanaxadiol and ginsenoside Rd among the saponin fraction were especially increased in the activity of hepatic antioxidative enzyme and decreased the lipid peroxidation that was expressed in term of MDA formation. This comprehensive antioxidant effects of red ginseng saponin seems to be by a certain action of saponin other than a direct antioxidant action.
Lavandula vera is indispensable member of the herb family, used for perfumes and potpourri. Androgens have profound effects on the physiology of the sebaceous gland. Sebum is secreted due to the effect of androgen, which starts to be secreted at puberty. Using the human sebocyte cell line SZ95. the author investigated the inhibitory effect of Lavandula vera on the lipid production. Light microscopic finding were examined numerous cytoplasmic lipid droplets SZ95 cells by Oil red staining and lipid droplets were increased markedly by testosterone. On the other hand, combined treatment with Lavandula vera and testosterone resulted in a lower lipid droplets than with testosterone alone in a dose-dependent manner. These findings indicate that Lavandula vera acts antagonistically to testosterone and inhibits the lipid synthesis of SZ95 cells at the cellular level.
The aim of this study was to investigate the effect of ethanol extract of Fagopyrum escuentum(FE) on the melanogenesis. To determine whether ethanol extract of FE suppress melanin synthesis in cellular level, B16F10 melanoma cells were cultured in the presence of different concentrations of FE ethanol extract. In the present study, the author examined the effects of FE ethanol extract on cell proliferation, melanin contents, tyrosinase activity. Cell proliferation was slightly increased by treatment with ethanol extract of FE (25-200 μg/ml). The ethanol extract of FE effectively suppressed melanin contents at a dose of 100 μg/ml. It was observed that the color of cell pellets was totally whitened compared with the control. The ethanol extract of FE inhibited tyrosinase activity, regulate melanin biosynthesis as the key enzyme in melanogenesis. These results suggest that the ethanol extract of FE exerts its depigmenting effects through the suppression of tyrosinase activity. And it may be a potent depigmetation agent in hyperpigmentation condition.
The development of random amplified polymorphic DNA (RAPD) and expressed sequence tag-derivedsimple sequence repeats (EST-SSRs) provided a useful tool for investigating Korean ginseng genetic diversity. In this study,18 polymorphic markers (7 RAPD and 11 EST-SSR) selected to assess the genetic diversity in 31 ginseng accessions (11Korean ginseng cultivars and 20 breeding lines). In RAPD analysis, a total of 53 unique polymorphic bands were obtainedfrom ginseng accessions and number of amplicons ranged from 4 to 11 with a mean of 7.5 bands. Pair-wise genetic similaritycoefficient (Nei) among all pairs of ginseng accessions varied from 0.01 to 0.32, with a mean of 0.11. On the basis of theresulting data, the 31 ginseng accessions were grouped into six clusters. As a result of EST-SSR analysis, 11 EST-SSR mark-ers detected polymorphisms among the 31 ginseng accessions and revealed 49 alleles with a mean of 4.45 alleles per primer.The polymorphism information content (PIC) value ranged from 0.06 to 0.31, with an average of 0.198. The 31 ginsengaccessions were classified into five groups by cluster analysis based on Nei’s genetic distances. Consequently, the results ofginseng-specific RAPD and EST-SSR markers may prove useful for the evaluation of genetic diversity and discrimination ofKorean ginseng cultivars and breeding lines.
Korean ginseng (P. ginseng C. A. Meyer) is one of the most important medicinal plant in the world. Understanding genetic variability among the assortment of Korean ginseng is important for breeding. The aim of this study was to molecularly characterize Korean ginseng cultivar and breeding lines through the use of eight previously reported STS markers (MFGp183, MFGp130, MFGp110, UFGp74, UFGp163, MFGp108, MFGp81 and UFGp156). All STS markers produced interpretable electropherograms from 31 accessions consisting of 11 Korean ginseng cultivars and 20 breeding lines. When eight STS markers were combined, we identified to total 19 genetic patterns; in particular, nine cultivars (Chunpoong, Yunpoong, Gopoong, Gumpoong, Sunpoong, Sunone, Cheongseon, Sunhyang, Cheonryang) and 5 breeding lines (G08012, G04079, G04075, G08036, G04110) in ginseng samples can be discriminated from the others. Together with other available markers, these STS markers will contribute to the management of ginseng genetic resources and the protection of breeders' rights.
This study describes the identification of Panax species using mitochondrial consensus primers. Initially, a total of thirty primers were tested in ten Korean ginseng cultivars and two foreign Panax species, P. quinquefolius and P. notoginseng. In the polymerase chain reaction (PCR) amplification results, three primers (cox1, nad1/2-3 and nad2/1-2) generated co-dominant polymorphic banding patterns discriminating Korean ginseng cultivars from P. quinquefolius and P. notoginseng. However, these primers could not generated polymorphisms among the Korean ginseng cultivars, and simply represented species-specific polymorphisms for P. quinquefolius and P. notoginseng. Primers PQ91 and PN418 were designed from the consensus sequence of nad1/2-3 region. Two banding patterns (A or B) were detected in PQ91. Korean ginseng cultivars and P. notoginseng shared the same banding pattern (A type) and P. quinquefolius was identified another banding pattern (B type). In the case of PN418, two banding patterns (A or B) were detected in the Korean ginseng cultivars and two foreign Panax species. Korean ginseng cultivars and P. quinquefolius shared the same banding pattern (A type) and P. notoginseng was identified another banding pattern (B type). The combination banding patterns of three Panax species, Korean ginseng cultivars (Panax ginseng C. A. Mey.), P. quinquefolius and P. notoginseng, was identified as 'AA', 'BA' and 'AB', respectively. Consequently, PQ91 and PN418 primer sets can be used to distinguish among Panax species.
This study was carried out to identify Korean ginseng cultivars using peptide nucleic acid (PNA) microarray. Sixty-seven probes were designed based on nucleotide variation to distinguish Korean ginseng cultivars of Panax ginseng. Among those PNA probes, three (PGB74, PGB110 and PGB130) have been developed to distinguish five Korean ginseng cultivars. Five Korean ginseng cultivars were denoted as barcode numbers depending on their fluorescent signal patterns of each cultivar using three probe sets in the PNA microarray. Five Korean ginseng cultivars, Chunpoong, Yunpoong, Gopoong, Gumpoong and Sunpoong, were simply denoted as '111', '222', '211', '221' and '122', respectively. This is the first report of PNA microarray which provided an objective and reliable method for the authentication of Korean ginseng cultivars. Also, the PNA microarray will be useful for management system and pure guarantee in ginseng seed.