• Title/Summary/Keyword: Cell-in-cell

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Enzymological Evaluation of Oral Inflammation inhibitory activity by Aloe vera peel extract (Aloe vera peel 추출물에 의한 구강염증 저해 효과의 효소학적 평가)

  • Pack, Chung-Sun;Ryu, Il-Hwan;Lee, Kap-Sang
    • Korean Journal of Food Science and Technology
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    • v.33 no.6
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    • pp.753-759
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    • 2001
  • To evaluate antiinflammation of Aloe vera peel, antiimflammation substances were extracted from Aloe vera peel and identified, and we investigated the effect of the its substance the inhibitory effect on the activity of hyaluoronidase, elastase, collagenase and prostaglandin endoperoxide synthase. The water extract from Aloe vera peel were successfully purified with solvent fractionation, silica gel column chromatography, preparative thin layer chromatography and UV spectrometer. Two purified active substances were identified as aloe-emodin and barbaloin by Mass Spectrometer, $^1H-NMR$ and FT-IR. Aloe-emodin and barbaloin. $IC_{50}$ values of aloe-emodin and barbaloin against hyaluronidase activity were 40 and $70\;{\mu}g/mL$, respectively. Leuckocyte elastase, which is related to the destruction of various tissue, $IC_{50}$ values of them were 50 and $60\;{\mu}g/mL$, respectively. $IC_{50}$ values of aloe-emodin and barbaloin against collagenase activity were 40 and $60\;{\mu}g/mL$, respectively. and $IC_{50}$ values of aloe-emodin and barbaloin aganist the prostaglandin endoperoxide synthase, which play an important role in inflammatory reactions, were 40 and $70\;{\mu}g/mL$, respectively. Inhibitory effects of aloe-emodin, barbaloin and aspirin against carrageenan paw edema were 74.9, 52.9 and 51.9% as inhibiton percentage, respectively, at dose of 100 mg/kg and that of indomethancin was 49.7 at dose of 10 mg/kg. Cell cytotoxicity of barbaloin against human gingival cells was lower than that of aloe-emodin. Aloe-emodin and barbaloin did not show cytotoxicity against human gingival cells at concentration of 1.0 and $5.0\;{\mu}g/mL$, However, aloe-emodin and barbaloin showed less cytotoxicity than chlorhexidine, which usually have been used as the agent of anticaries and antiinflammation. These results suggested that aloe-emodin and barbaloin from Aloe vera peel have the effect of anticaries and antiinflammation.

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Preservation of Washed Fresh Ginsengs by Gamma Irradiation (방사선(放射線) 조사(照射)에 의한 수세된 수삼(水蔘)의 저장(貯藏))

  • Cho, Han-Ok;Byun, Myung-Woo;Kwon, Joong-Ho;Lee, Jae-Won
    • Applied Biological Chemistry
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    • v.29 no.3
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    • pp.288-293
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    • 1986
  • The washed fresh ginsengs packed with air, vaccum and nitrogen gas were irradiated at the levels of 1,2 and 3kGy gamma radiation and then stored at $4{\sim}5^{\circ}C$ for 90days to investigate the effects of gamma radiation on microbial inactivation, eelworm disinfestation and physicochemical changes. After a 90 day storage, $2{\sim}3kGy$ irradiated groups showed 20% of weight loss and 10% of rot while non-irradiated group 100% and 20% or more, respectively. Also the irradiated groups showed somewhat lower values of specific gravity, color density and hardness immediately after irradiation, thereafter higher value of them with storage time than those of non-irradiated group. The irradiation increased the yields of ginseng extract and crude saponins but no effects on the proximate composition and TLC and HPLC patterns of saponin. The food-borne microorganisms decreased in viable cell counts by $2{\sim}3$ log cycles with $2{\sim}3kGy$ radiation and the eelworms were completely disinfested with 1 kGy radiation.

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Combining Ginsenoside F1 with (-)-Epigallocatechin Gallate Synergistically Protects Human HaCaT Keratinocytes from Ultraviolet B-Induced Apoptosis (Ginsenosdie F1과 EGCG의 상승작용에 의한 자외선조사에 의한 세포 사멸 방지)

  • Tae Ryong, Lee;Si Young, Cho;Eun Hee, Lee;Myeong Hoon, Yeom;Ih-Seop, Chang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.253-261
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    • 2004
  • Ginsenosides and green tea extracts show a variety of biomedical efficacies such as anti-aging, anti-oxidation and anti-tumor-promotion effects. (-)-Epigallocatechin-3-gallate (EGCG) has been reported to inhibit the UVB-induced apoptosis by increasing the Bcl-2-to-Bax ratio. We have previously shown that ginsenoside Fl protects human HaCaT cells from ultraviolet-B (UVB)-induced apoptosis by maintaining constant levels of Bcl-2 and Brn-3a. Here, we investigate the combined effect of ginsenoside Fl and EGCG on the protection of human HaCaT keratinocyte against UVB-induced apoptosis. When treated individually, although 5 ${\mu}$M ginsenoside Fl and 50${\mu}$M EGCG protected cells from UVB-induced apoptosis, 2${\mu}$M ginsenoside Fl or 10${\mu}$M EGCG treatment showed very little protection effect. However, cotreatement of 2${\mu}$M ginsenoside Fl and 10${\mu}$M EGCG successfully protected HaCaT cells from UVB-induced cell death. As expected, combining ginsenoside Fl and EGCG efficiently prevented UVB-induced decrease of Bcl-2 and Brn-3a expression. In addition, cotreatment with ginsenoside F1 and EGCG prevented the dephosphorylation of Rb, whereas individual treatment with ginsenoside Fl or EGCG failed to prevent the dephosphorylation of Rb even at high concentrations.

Effects of Ginsenosides on Glucose Uptake and Insulin Secretion

  • Park, Min-Woo;Shin, Eun-Jung;Ko, Sung-Kwon;Chung, Sung-Hyun
    • Proceedings of the Ginseng society Conference
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    • 2007.05a
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    • pp.23-24
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    • 2007
  • Purpose: 인삼이 항당뇨 활성을 가진다는 연구가 많은 연구자들에 의해 진행되었고, 이는 인삼의 구성 성분 중 ginsenoside에 기인한다는 보고가 있다. 본 연구는 ginsenoside의 항당뇨 작용기전을 in vitro에서 알아보고자 3T3-L1 지방세포에서 glucose uptake와 췌장 베타세포인 HIT-T15 세포에서 insulin 분비 효과를 확인하였다. 이를 위하여 인삼을 식초로 처리한 긴삼의 70% MeOH 분획으로부터 protopanaxadiol 계인 ginsenoside $Rb_2$, $Rg_3$ 그리고 protopanaxtriol 계인 $Rg_2$를 분리하여 본 실험에 사용하였다. Method: Ginsenoside $Rb_2$, $Rg_2$, $Rg_3$가 지방 세포에서 glucose uptake에 미치는 효과를 확인하기 위하여 3T3-L1 세포를 DMEM (Dulbecco's Modified Eagle's Medium) 배지에서 분화 유도시켰으며 3T3-L1 preadipocyte가 80% 정도 자라면 분화 유도 배지 (5% fetal bovine serum (FBS), 0.5 mM isobutylmethylxanthine (IBMX), 1 mM dexamethasone 그리고 $10{\mu}g/ml$ insulin가 포함된 DMEM)로 4일, $10{\mu}g/ml$ insulin가 포함된 DMEM으로 2일, FBS만 포함된 DMEM으로 2일 배양하여 총 8일 동안 분화를 유도하였다. 분화 유도된 3T3-L1 adipocytes 에 각각 $Rb_2$, $Rg_2$, $Rg_3$$20{\mu}M$로 처리하여 16시간 배양하여 low glucose DMEM에서 3시간 배양한 후에 $37^{\circ}C$에서 insulin 10 ng/ml 과 각각 $Rb_2$, $Rg_2$, $Rg_3$가 포함된 Krebs Ringer Hepes buffer(KRP buffer)에서 20분간 배양하였다. 2-deoxy-D-[$^3H$]-glucose를 넣고 10분 후에 차가운 PBS로 반응을 종결시켜 lysis buffer로 cell을 모은 후 scintillation counter를 이용하여 glucose를 측정하였다. Insulin 분비 효과는 HIT-T15 세포와 일차 배양한 흰쥐 소도세포(islets)를 사용하여 확인하였다. HIT-T15 세포는 24 well plate에 well 당 $2{\times}10^5$ 개씩 분주하여 24시간 동안 배양한 후 시료를 처리하였으며 소도 세포는 Sprague-Dawley rat의 췌장에 collagenase가 포함된 Hanks' Balanced Salt Solution(HBSS)을 주입하여 분리하고 islets을 얻었다. 분리한 소도세포를 $1{\sim}2$일 동안 배양하여 $Rb_2$, $Rg_2$, $Rg_3$가 각각 $20{\mu}M$의 농도로 첨가된 insulin 측정용 buffer인 Krebs-Ringer buffer (KRB+0.3% BSA, KRBB)에 $37^{\circ}C$에서 1시간 incubation 시킨 후 배양액으로 분비된 인슐린의 양을 측정하였다. 한편 ginsenoside의 인슐린 분비 촉진 기전을 알아보기 위한 실험에서는 ATP-sensitive $K^+$ channel opener인 diazoxide (0.5 mM)가 ginsenoside에 의해 촉진된 인슐린 분비를 억제하는지 살펴보았다. Result: glucose uptake assay 에서는 $Rg_2$가 가장 크게 glucose uptake를 증가시켰고 $Rb_2$, $Rg_3$는 그 활성이 크지 않았다. 한편 Insulin 분비 효과는 diol계인 $Rg_3$에서 용량 의존적으로 인슐린의 분비를 촉진시켰으며 $20{\mu}M$ 농도에서 대조군과 비교해 1.5배 이상의 분비 촉진 효과를 보였고 triol계인 $Rg_2$ 에서는 이러한 효과가 나타나지 않았다. $Rg_3$의 인슐린 분비 촉진 기전을 0.5 mM 의 diazoxide를 이용하여 확인한 결과 $Rg_3$에 의해 촉진된 인슐린 분비를 감소시켰다. 이로 미루어보아 $Rg_3$의 인슐린 분비 촉진 기전은 ATP-sensitive $K^+$ 채널의 봉쇄에 의한 것임을 확인할 수 있었다.

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Bioleaching of Mn(II) from Manganese Nodules by Bacillus sp. MR2 (Bacillus sp. MR2에 의한 망간단괴의 생물용출)

  • Choi, Sung-Chan;Lee, Ga-Hwa;Lee, Hong-Keum
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.411-415
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    • 2009
  • Some microorganisms are capable of leaching Mn(II) from nonsulfidic manganese ores indirectly via nonenzymatic processes. Such reductive dissolution requires organic substrates, such as glucose, sucrose, or galactose, as a source of carbon and energy for microbial growth. This study investigated characteristics of Mn(II) leaching from manganese nodules by using heterotrophic Bacillus sp. strain MR2 provided with corn starch as a less-expensive substrate. Leaching of Mn(II) at 25.6 g Mn(II) $kg^{-1}$ nodule $day^{-1}$ was accompanied with cell growth, but part of the produced Mn(II) re-adsorbed onto residual $MnO_2$ particles after 24 h. Direct contact of cells to manganese nodule was not necessary as a separation between them with a dialysis tube produced similar amount [24.6 g Mn(II) $kg^{-1}$ nodule $day^{-1}$]. These results indicated an involvement of extracellular diffusible compound(s) during Mn(II) leaching by strain MR2. In order to optimize a leaching process we tested factors that influence the reaction, and the most efficient conditions were $25\sim35^{\circ}C$, pH 5~7, inoculum density of 1.5~2.5% (v/v), pulp density of 2~3 g/L, and particle size <75 ${\mu}m$. Although Mn(II) leaching was enhanced as particle size decrease, we suggest <212 ${\mu}m$ as a proper size range since more grinding means more energy consumption The results would help for the improvement of bioleaching of manganese nodule as a less expensive, energy-efficient, and environment-friendly technology as compared to the existing physicochemical metal recovery technologies.

Studies on the Estimation of Pregnancy Rate of White Hanwoo (Albino Korean Native Cattle) with Sexed Semen (백한우의 성판별 정액을 이용한 수태율 추정에 관한 연구)

  • Kim, Sung Woo;Choi, Jinseok;Choe, Changyoung;Kim, Dongkyo;Ko, Yeoung-Gyu;So, Chungsil;Seong, Hwan-Hoo
    • Journal of Embryo Transfer
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    • v.30 no.1
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    • pp.45-50
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    • 2015
  • The purpose of this study is to produce wanted sex progeny of genetically confined White Hanwoo (albinism) with preselected sex sperm. One bull of White Hanwoo was chosen for semen donor and X sperm was sorted by MoFlo XDP cell sorter. To compare the pregnancy and birth rates, KPN straw was used as control, total number of unsorted sperm was $20{\times}10^6/straw$. Sexed X frozen semen with $20{\times}10^6$ cells or $4{\times}10^6$ cells per straw were in seminated twice on Hanwoo heifers. The abnormality of the sexed X semen was $24.9{\pm}7.31%$ and distal reflex abnormality of mid piece was significantly (p<0.05) higher (11.7%) compared with that of KPN 768 (5.6%). There were no differences on the pregnancy and birth rates between $2{\times}10^6$ cells or $4{\times}10^6$ cells of X-sperm but KPN semen showed significant differences (p<0.05). The pregnancy rates of KPN 768, $2{\times}10^6$ cells and $4{\times}10^6$ cells X-sperm of White Hanwoo cattle were 85.0%, 26.3% and 50%. The birth rates were 80.0%, 15.8% and 21.4%, respectively. The female offspring rates of KPN 768, $2{\times}10^6$ cells and $4{\times}10^6$ cells X-sperm of White Hanwoo cattle were 43.8%, 100% and 100% (p<0.05). These results indicated that sex sorted White Hanwoo could be used for the production of wanted progeny with $2{\times}10^6$ cells/straw for AI. To increase the efficiency of calf production, the sperm number of sex sorted semen will be optimized for sex selection of White Hanwoo progeny.

Radiotherapy Results of Uterine Cervix Cancer Stape IIB : Overall Survival, Prognostic factors, Patterns of Failure and Late Complications (자궁경부암 병기 IIB에서의 방사선 치료 성적: 생존율 및 예후인자, 치료 후 실패양상, 만성 합병증)

  • Kim Eun-Seog;Choi Doo-Ho;Huh Seung-Jae
    • Radiation Oncology Journal
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    • v.16 no.1
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    • pp.51-61
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    • 1998
  • Purpose : Treatment of choice for uterine cervix cancer stage IIB is radiotherapy. We analyzed survivals, Prognostic factors, patterns of failure and complications. Materials and Methods : This is a retrospective analysis of 167 patients with stage IIB carcinoma of uterine cervix treated with curative external pelvic and high dose rate intracavitary radiotherapy at the Department of Therapeutic Radiology, Soonchunhyang University Hospital from August 1985 to August 1994. All the patients followed up from 3 to 141 months(mean 60 months) and age of patients ranged from 31 to 78 years at presentation(mean : 55 years). Results : Overall complete response rate was $84\%$. The response rate for squamous cell carcimoma and adenocarcinoma were $86\%$ and $60\%$, respectively. Overall 5-years survival rate and disease free survival rate was 62 and $59\%$, respectively Mass size and treatment response were significant Prognostic factors for survival Pathologic type and parametrial involvement were marginally significants Prognostic factors. Local failure was 43 cases, distant metastasis was 14 cases and local failure plus distant metastasis was 3 cases, and most of local failures occurred within 24 months, distant metastasis within 12 months after treatment Twenty eight($16.8\%$) patients developed late rectal and urinary complications There were tendency to increasing severity and frequency according to increased fractional dose and total(rectal and bladder) dose. Conclusions : Survival rate was significantly related to tumor size and radiotherapy response. Tumor size should be considered in the clinical s1aging. To increased survival and local control, clinical trials such as decreasing duration of radiotherapy or addition of chemotherapy is needed. To detect early recurrence, regular follow up after RT is important. Because total rectal and bladder dose affected late complications. meticulous vaginal packing is needed to optimize dose of normal tissues and to decrease late complications.

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Isolation and Characterization of Elastase Inhibitor from Areca catechu (빈랑으로부터 Elastase 저해물질의 분리 및 특성조사)

  • 조중제;이건국;조병기;최정도
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.26 no.1
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    • pp.163-186
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    • 2000
  • We have previously screened 150 medicinal plants on the inhibition of elastase and found a significant inhibitory effects of the extracts of Areca catechu L. on the aging and inflammation against the skin tissues. To isolate and identify the compounds having biological activity, we was further purified by each of the solvent fractions, silica gel column chromatography, preparative TLC and reversed-Phase HPLC. Peak in HPLC, which coincided with the inhibitory activity against elastase, was identified as Phenolic substance using various colorimetric methods, UV, and IR. $IC_{50}$/ values of phenolic substance purified from Areca catechu were 26.9 $\mu\textrm{g}$/$m\ell$ for porcine pancreatic elastase (PPE) and 60.8 $\mu\textrm{g}$/$m\ell$ for human neutrophil elastase (HNE). This Phenolic substance showed more potent activity than those of reference compounds, oleanolic acid (76.5 $\mu\textrm{g}$/$m\ell$ for PPE, 219.2 $\mu\textrm{g}$/$m\ell$ for HNE) and ursolic acid (31.0 $\mu\textrm{g}$/$m\ell$ for PPE, 118.6 $\mu\textrm{g}$/$m\ell$ for HNE). According to the Lineweaver-Burk Plots, the inhibition against both PPE and HNE by this phenolic substance was competitive with substrate. Phenolic substance from Areca catechu exhibited high free radical scavenging effect ($SC_{50}$/ : 6 $\mu\textrm{g}$/$m\ell$) and inhibited effectively hyaluronidase activity ($IC_{50}$/: 210 $\mu\textrm{g}$/$m\ell$). These results suggest that the Phenolic substance Purified from Areca catechu showed anti-aging effect by protecting connective tissue proteins.

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Quality Characteristics and Inhibition Activity against Helicobacter pylori KCCM 40449 of Liquorice Yogurts Manufactured by Exopolysaccharide Producing Lactic Acid Bacteria (Exopolysaccharide 생성 유산균을 이용한 감초 추출물 첨가 Yoghurt의 품질특성 및 Helicobacter pylori KCCM 40449 억제활성)

  • Jung, Seung-Won;Kim, Cheol Woo;Lee, Su Han
    • Food Engineering Progress
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    • v.15 no.4
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    • pp.346-354
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    • 2011
  • This study was carried out to fortify the antimicrobial activity of yoghurt by adding liquorice extract to it. The liquorice extracts (1 mg/mL) showed relatively high antibacterial activity against H. pylori KCCM 40449 (p < 0.05). The solvent liquorice extracts of minimal inhibitory concentrations (MIC) against H. pylori KCCM 40449 were 25- 100 ${\mu}g$/mL. Lactobacillus amylovorus DU-21 with high EPS production ability were inoulated to milk after the addition of different amounts of liquorice extracts (0.0%, 0.05%, 0.1% and 0.2%). The physico-chemical characteristics of yoghurts added with liquorice extracts were examined. The initial pH, titratable acidity, viscosity and viable cell counts of the yoghurt added liquorice extracts were 3.41-3.51, 1.021-1.091%, 1,686-1,930 cp and 9.41-9.38 Log CFU/mL, respectively. The viscosity and syneresis of yoghurt were better than that of the control. Antimicrobial activity against H. pylori KCCM 40449 increased with increasing addition of liquorice extract. However, the sensory score of yoghurt added with different amounts of liquorice extracts was lower than that of the control (p < 0.05). As a result of the sensory evaluations, the flavor, taste, texture, color and overall acceptability of the yoghurt with 0.05% liquorice extract were found to be much better than those of the other groups (p < 0.05). Overall, the optimal amount of liquorice extract added in the manufacture of yoghurt was 0.05% of the total weight. Further studies on increment of antimicrobial activity and palatability of liquorice extract added yoghurt are necessary.

Whitening Activity of Sambucus Sieboldiana Var. Pendula (Nakai) Extract (말오줌나무 추출물의 미백활성 검증)

  • Yoo, Dan-Hee;Kim, Jin-Tae;Oh, Min-Jeong;Yeom, Hyeon-Ji;Lee, Jin-Young
    • Journal of Life Science
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    • v.29 no.3
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    • pp.279-286
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    • 2019
  • This study evaluated the anti-oxidant and whitening effects of a 70% ethanol extract of the Sambucus sieboldiana var. pendula (Nakai) (SS). At $1,000{\mu}g/ml$ concentration, the electron donating ability of this SS extract was found to be 86.21% and the ABTS+ radical scavenging ability was 97.9%. In terms of whitening activity, the tyrosinase inhibitory effect of the extract was 37%, also at $1,000{\mu}g/ml$ concentration. To explore the extractefftoxicity to B16F10 melanoma cells, a 3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide assay was performed. Results showed 90% or more cells remained viable at $100{\mu}g/ml$ concentration. A Western blot of the SS extract was used to measure microphthalmia-associated transcription factor (MITF), tyrosinase-related protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2), and the tyrosinase protein expression inhibitory effect at 25, 50, $100{\mu}g/ml$ concentrations; ${\beta}-actin$ was used as a positive control. Consequently, the MITF, TRP-1, TRP-2, and the tyrosinase protein expression inhibitory effect were seen to decrease by 34.5%, 45.6%, 58.4%, and 79.6%, respectively, at $100{\mu}g/ml$ concentration. These were also then measured by reverse transcription-polymerase chain reaction at 25, 50, $100{\mu}g/ml$ concentrations with GAPDH as a positive control. As a result, the SS extract was seen to decrease MITF, TRP-1, TRP-2, and the tyrosinase protein expression inhibitory effect by 85.4%, 67.5%, 85.2%, 67.1%, respectively at the $100{\mu}g/ml$ concentration. We therefore confirmed the possibility of Sambucus sieboldiana var. pendula (Nakai) extract as a whitening material.