Pheophorbide a-mediated photodynamic therapy induces autophagy and apoptosis via the activation of MAPKs in human skin cancer cells

YOON, HYO-EUN,OH, SEONE-HEE,KIM, SOO-A,YOON, JUNG-HOON,AHN, SANG-GUN Spandidos Publications 2014 Oncology reports Vol.31 No.1

Pheophorbide a (Pa), a chlorophyll derivative, is a photosensitizer that can induce significant antitumor effects in several types of tumor cells. The present study investigated the mechanism of Pa-mediated photodynamic therapy (Pa-PDT) in the human skin cancer cell lines A431 and G361. PDT significantly inhibited the cell growth in a Pa-concentration-dependent manner. We observed increased expression of Beclin-1, LC3B and ATG5, which are markers of autophagy, after PDT treatment in A431 cells but not in G361 cells. In G361 cells, Pa-PDT strongly induced PARP cleavage and subsequent apoptosis, which was confirmed using Annexin V/Propidium iodide double staining. Pa-PDT predominantly exhibited its antitumor effects via activation of ERK1/2 and p38 in A431 and G361 cells, respectively. An in vivo study using the CAM xenograft model demonstrated that Pa-PDT strongly induced autophagy and apoptosis in A431-transplanted tumors and/or apoptosis in G361-transplanted tumors. These results may provide a basis for understanding the underlying mechanisms of Pa-PDT and for developing Pa-PDT as a therapy for skin cancer.

Clostridium difficile에 의한 설사의 예후인자

김준형,김희정,구남수,김영근,최준용,신소연,박윤선,김연아,김명수,정수진,최희경,송영구,이경원,김준명 대한감염학회 2007 감염과 화학요법 Vol.39 No.2

배경 : Clostridium difficile에 의한 설사(Clostridium difficile associated diarrhea; CDAD)는 임상 경과가 다양하다. 최근에 유럽과 북미에서 C. difficile 감염의 발생률이 증가하고 고전적 치료에 잘 반응하지 않으며 이환률이 증가하였고, 이러한 원인이 새로운 균주의 탄생에 기인한다고 보고되었다. CDAD의 예후에 영향을 미치는 세균성 요인과 숙주 요인을 확인하기 위해 본 연구를 진행하였다. 재료 및 방법 : 2002년 8월부터 2003년 12월까지 CDAD가 진단된 20세 이상인 환자들을 대상으로 후향적 Cohort 연구를 하였다. 세균성 요인을 확인하기 위해 cdt A, cdtB, tcd A, tcd A rep 그리고 tcd B 유전자(binary toxin)를 확인하였다. 설사가 치료 시작 후 11일 이상 지속되거나, 2달 이내에 재발하거나, 수술 혹은 다른 시술이 필요한 경우, 사망한 경우 예후가 좋지 않은 것으로 정의하였다. 결과 : 총 115예에서 toxin을 생성하는 C. difficile가 동정되었으며, Toxin A와 toxin B 모두 양성인 균이 91예, toxin B만 양성인 균이 24예였다. Toxin A 생성 여부가 예후에 영향을 미치지 않았지만, 제산제를 사용한 환자에서 그렇지 않은 환자에 비해 toxin B만 양성인 균이 많이 동정되었다(P<0.05). 예후가 좋지 않은 경우는 39예(33.9%)였고 76예(66.1%)에서 예후는 양호하였다. 단변량 분석에서 70세 이상의 고령, 남성, 증상 발현 후 사용한 항생제의 개수 사용, 증상 발현 후 carbapenem, aminoglycoside, glycopeptide 사용, 당뇨 및 뇌졸중 병력이 있는 경우, 그리고 높은 Charlson index가 불량한 예후 인자로 확인되었다. 그러나 독립적인 예후 인자를 조사했을 때에는70세 이상의 고령(odds ratio=3.378, P=0.009), 증상 발현후 carbapenem 사용(odds ratio 7.210, P<0.001)이 예후에 중요한 영향을 미치는 요소로 확인되었다. 결론 : 70세 이상의 고령과 증상 발현 후 carbapenem 사용이 CDAD 독립적인 불량한 예후인자이다. Background : Clostridium difficile associated diarrhea (CDAD) has a wide range of clinical manifestations. The prognostic factors of CDAD are not fully understood. Materialsand Methods : A retrospective cohort study of 115 patients with CDAD from Aug. 2002 to Dec. 2003 was conducted to evaluate prognostic factors of CDAD. Bacteriologic factors were determined by detecting the binary toxin gene, tcd A, tcd A rep and tcd B gene. Poor prognosis was defined as diarrhea more than 10 days even with classic treatment, recurrence, death, and moribund discharge. Results : Approximately 79% of isolated strains were toxin A+/B+ strains and 21% were toxin A-/B+ strains. There was no difference in prognosis between toxin A+ and toxin A- strains. 39 (33.9%) cases showed poor prognosis and 76 (66.1%) cases showed good prognosis. Univariate analyses revealed that the poor prognostic factors were old age over 70 years old, male, the number of antibiotics used after onset of symptom, the administration of carbapenems, aminoglycosides, glycopeptides after onset of symptom, history of DM and stroke, and high Charlson comorbidity index. Multiple logistic regression analysis identified old age over 70 years old (odds ratio=3.378, P=0.009) and the administration of carbapenems after onset of symptom (odds ratio 7.210, P<0.001) as the independent poor prognostic factors. Conclusion : Old age over 70 and the administration of carbapenems after onset of symptom were the poor prognostic factors for CDAD caused by none-binary toxin producing strains.

수입 및 국내 유통 식품 중의 Ochratoxin A에 관한 조사연구

이동호,강민철,이선화,정동윤,김재이,김형수,김은정,유병옥,김연주,정순아,서영선,김인복,홍무기 식품의약품안전청 2001 식품의약품안전청 연보 Vol.5 No.-

곰팡이 독소 중 현재 식품골전에 r'1 등재되어 있는 ochratoxin A의 규제기준 설정에 대한 기초자료로 활용하기 위한 방안의 일찬으로 곡류, 두류, 견과류 및 장류에 있어서 ochratoxin A의 오염정도를 조사하였다. 실헐방법으로 1차 screening은 ochratoxin test t를, 최종확인은 irnrrlunoaffihie· column을 이용한 HPLC 분석법을 사용하였파. 회수율은 수수를 제외하고 모두 80%이상을 나타내었고 최저겊출 한계치는 0.Sppb였다. 총 121건(보리 9잔, 찹쌀 7건, 백미 8건, 조 9건, 수수10건, 서리태 11건, 녹두 3건, 백태 9건, 동분 5건, 팥 7건, 강낳콩 8건, 땅콩 7건, 호두 7건, 혼합장 11건, 청국장 10건)에 대하여 조사한 결과 1차 screeRing에서 39건의 양성반응을 보였으나 HPLC로 최종 확인 결과 모든 시료에서 불검출 되었다

Litsenolide A2: The major anti-inflammatory activity compound in <i>Litsea japonica</i> fruit

Ham, Young-Min,Cho, Su-Hyeon,Song, Sang-Mok,Yoon, Seon-A,Lee, Yong-Bum,Kim, Chang-Sook,Kwon, Seung-Hae,Jeong, Myeong Seon,Yoon, Weon-Jong,Kim, Kil-Nam Elsevier 2017 Journal of Functional Foods Vol.39 No.-

<P><B>Abstract</B></P> <P>In this study, we investigated the anti-inflammatory effect of litsenolide A2 (LNA2) isolated from <I>L. japonica</I> fruit and its mechanism of action in LPS-induced RAW 264.7 cells. LPS-induced production of anti-inflammatory mediators and cytokines such as NO, PGE<SUB>2</SUB>, iNOS, COX-2, TNF-α, and IL-6 was significantly inhibited by LNA2. Next, western blot experiments were performed to investigate the mechanism of action of the anti-inflammatory effect of LNA2. The results indicated that LNA2 markedly reduced the LPS-induced activation of NF-κB and the nuclear translocation of NF-κB p65. Furthermore, LNA2 also inhibited the phosphorylation of MAPK by LPS. LNA2 was the major component in the ethanol extract of <I>L. japonica</I> at 59 mg/g according to high performance liquid chromatography (HPLC). These results indicate that LNA2 can be used as a functional and marker compound for standardization of the manufacturing process when <I>L. japonica</I> fruit is used as a functional food.</P> <P><B>Highlights</B></P> <P> <UL> <LI> We study the anti-inflammatory effect of LNA2 in LPS-activated RAW 264.7 cells. </LI> <LI> LNA2 inhibited production of pro-inflammatory mediator and cytokines. </LI> <LI> LNA2 exhibited anti-inflammatory properties by suppression of NF-κB and MPAKs. </LI> <LI> LNA2 was the major component in the ethanol extract of <I>L. japonica</I> at 59 mg/g. </LI> </UL> </P>

Dimethoate의 뿔가위벌(Osmia pedicornis) 급성 접촉 및 섭식독성평가

전경미(Kyongmi Chon),김주영(Juyeong Kim),김보선(Bo-Seon Kim),윤창영(Chang-Young Yoon),오진아(Jin-A Oh),박홍현(Hong-Hyun Park),윤형주(Hyung-Joo Yoon) 한국농약과학회 2021 농약과학회지 Vol.25 No.4

Honeybee (Apis mellifera) has been used as a protective surrogate in the regulatory risk assessment processes. Recently, EFSA proposed the inclusion of two additional surrogate species (Bombus spp., Osmia spp.) into EU pesticide risk assessment due to the global decline of pollinator populations and bee sensitivity difference. We investigated the acute contact and oral toxicity of dimethoate towards the male Osmia pedicornis based on the test guideline developed by the International Commission of Plant-Pollinator Relationships (ICP-PR) non-Apis group. The following dimethoate doses were used to determine the LD<SUB>50</SUB> for the acute contact toxicity test: 0.125, 0.25, 0.5, 1.0, 2.0 μg a.i./bee. Thirty newly emerged male bees per treatment groups were selected for the test and each bee was applied with 2 μL treatment solution on the dorsal side of the thorax. The acute contact toxicity of dimethoate 48 h and 96 h LD<SUB>50</SUB> for male Osmia pedicornis were 0.186 and 0.093 μg a.i./bee, respectively. Five dimethoate doses (0.017, 0.05, 0.15, 0.45, 1.35 μg a.i./bee) were used to determine the LD<SUB>50</SUB> for the acute oral toxicity test. Thirty replicates per treatment were used and each bee was housed individually and provided with 20 μL of aqueous 50% sucrose solution containing the test chemical. The acute oral toxicity of dimethoate 48 h and 96 h LD<SUB>50</SUB> for male Osmia pedicornis were 0.119 and 0.104 μg a.i./bee, respectively. In the near future, this test method could be used to assess the effect of various pesticides including neonicotinoids on Osmia species.

삼채의 조추출물과 유기용매 분획물에 대한 항산화 및 항염증 효과

이용범(Yong-Bum Lee),함영민(Young-Min Ham),윤선아(Seon-A Yoon),오대주(Dae-Ju Oh),송상목(Sang-Mok Song),홍인철(In-Choel Hong),이시택(Si Taek Lee),현호봉(Ho Bong Hyun),김창숙(Chang-Suk Kim),윤원종(Weon-Jong Yoon) 한국식품영양과학회 2017 한국식품영양과학회지 Vol.46 No.1

본 연구는 삼채의 조추출물과 유기용매 분획물들을 가지고 DPPH 라디칼 소거 활성에 의한 항산화 활성 검색 결과 디클로로메탄(CH2Cl2) 분획물과 에틸아세테이트(EtOAc) 분획물에서 라디칼 소거 활성을 나타냈으며, xanthine oxidase 억제 효과는 DPPH 활성 라디칼 소거 활성에서 제일 뛰어났던 에틸아세테이트 분획물에서, superoxide 소거 활성은 헥산(n-hexane) 분획물에서 활성이 나타났다. RAW 264.7 세포에 lipopolysaccharide로 자극을 주고 삼채 주정 추출물 및 유기용매 분획물들을 처리하여 확인해본 결과, 조추출물과 물 분획물을 제외한 나머지 유기용매 분획물에서 염증 유발 인자(NO, PGE2, iNOS, COX-2, IL-6 및 IL-1β) 생성억제 효과가 나타났으며, 그중 디클로로메탄 분획물과 에틸아세테이트 분획물에서 억제 효과를 확인할 수 있었다. 본 실험 결과 삼채 조추출물과 유기용매 분획물에서 항산화 효과 및 염증 유발 인자의 생성 억제 효과가 나타났으며, 이러한 결과 삼채에서 유효성분 추출을 통한 항산화, 항염증 물질의 연구 또는 예방하거나 치료할 수 있는 염증 억제 성분의 분리 및 그 작용기전 연구에 중요한 기초 자료가 될 것이라 생각한다. This study describes the preliminary evaluation of antioxidant and anti-inflammatory activities of Allium hookeri. A. hookeri was extracted using crude extract and then fractionated sequentially with n-hexane, CH2Cl2, EtOAc, and n-BuOH. To screen for antioxidant and anti-inflammatory agents effectively, we first examined the inhibitory effect of A. hookeri extracts on production of oxidant stresses (2,2-diphenyl-1-picrylhydrazyl, xanthine oxidase, and superoxide). In addition, we examined the inhibitory effects of A. hookeri on production of pro-inflammatory factors (nitric oxide, prostaglandin E2, inducible nitric oxide synthase, and cyclooxygenase-2) in murine macrophage RAW 264.7 cells stimulated with lipopolysaccharide. Of the sequential solvent fractions of A. hookeri, EtOAc fractions showed decreased production of oxidant stresses, and CH2Cl2 and EtOAc fractions of A. hookeri inhibited production of pro-inflammatory factors. EtOAc fraction inhibited production of pro-inflammatory cytokines (interleukin-6 and -1β). These results suggest that A. hookeri has significant effects on oxidant stresses and pro-inflammatory factors and is a possible antioxidant and anti-inflammatory therapeutic and preventive material.

Cariporide Enhances the DNA Damage and Apoptosis in Acid-tolerable Malignant Mesothelioma H-2452 Cells

Lee, Yoon-Jin,Bae, Jin-Ho,Kim, Soo-A,Kim, Sung-Ho,Woo, Kee-Min,Nam, Hae-Seon,Cho, Moon-Kyun,Lee, Sang-Han Korean Society for Molecular and Cellular Biology 2017 Molecules and cells Vol.40 No.8

The $Na^+/H^+$ exchanger is responsible for maintaining the acidic tumor microenvironment through its promotion of the reabsorption of extracellular $Na^+$ and the extrusion of intracellular $H^+$. The resultant increase in the extracellular acidity contributes to the chemoresistance of malignant tumors. In this study, the chemosensitizing effects of cariporide, a potent $Na^+/H^+-exchange$ inhibitor, were evaluated in human malignant mesothelioma H-2452 cells preadapted with lactic acid. A higher basal level of phosphorylated (p)-AKT protein was found in the acid-tolerable H-2452AcT cells compared with their parental acid-sensitive H-2452 cells. When introduced in H-2452AcT cells with a concentration that shows only a slight toxicity in H-2452 cells, cariporide exhibited growth-suppressive and apoptosis-promoting activities, as demonstrated by an increase in the cells with pyknotic and fragmented nuclei, annexin V-PE(+) staining, a $sub-G_0/G_1$ peak, and a $G_2/M$ phase-transition delay in the cell cycle. Preceding these changes, a cariporide-induced p-AKT down-regulation, a p53 up-regulation, an ROS accumulation, and the depolarization of the mitochondrial-membrane potential were observed. A pretreatment with the phosphatidylinositol-3-kinase (PI3K) inhibitor LY294002 markedly augmented the DNA damage caused by the cariporide, as indicated by a much greater extent of comet tails and a tail moment with increased levels of the p-histone H2A.X, $p-ATM^{Ser1981}$, $p-ATR^{Ser428}$, $p-CHK1^{Ser345}$, and $p-CHK2^{Thr68}$, as well as a series of pro-apoptotic events. The data suggest that an inhibition of the PI3K/AKT signaling is necessary to enhance the cytotoxicity toward the acidtolerable H-2452AcT cells, and it underlines the significance of proton-pump targeting as a potential therapeutic strategy to overcome the acidic-microenvironment-associated chemotherapeutic resistance.

Biological Activities of Immature Persimmon (Diospyros kaki) Extracts as Natural Functional Resource

Seon-A Yoon,Young-Min Ham,Dae-Ju Oh,Weon-Jong Yoon 한국식품영양과학회 2018 한국식품영양과학회 학술대회발표집 Vol.2018 No.10

TGF-β1 Expression by Proliferated Keratinocytes in the Skin of E-Irradiated Mice

A-Ran Yoon(윤아란),Do Nyun Kim(김도년),Min Koo Seo(서민구),Sang Taek Oh(오상택),Jung Seon Seo(서정선),Se Mo Jun(전세모),Jung-Ho Cha(차정호),Seungdeok Lee(이승덕),Suk Kyeong Lee(이숙경) 한국생명과학회 2012 생명과학회지 Vol.22 No.2

우리는 방사선피부염 동물 모델을 확립하여, 이차 면역 기관에서의 면역 세포 비율 변화를 관찰하였다. 또한, 방사선 조사에 의한 병소에서 transforming growth factor-β1 (TGF-β1)과 interlukin-10 (IL-10)의 발현을 증가시킨 세포를 분석하였다. Hairless-1 (HR-1) 쥐의 posterior dorsal 부위에 6 일간 매일 10 Gy 씩 electron (E)-ray를 국부 조사하여 방사선피부염 모델을 만들었다. FACS를 이용하여 면역 세포 비율의 변화를 분석한 결과 비장과 림프절에 존재하는 항원제시세포와 T 세포 및 B 세포들의 비율이 E-irradiation에 의해 영향을 받았다. 피부에서 세포 특이적인 마커와 사이토카인들의 발현 양상은 면역형광염색법으로 확인하였다. 방사선 조사 후, TGF-β1과 interlukin-17 (IL-17)은 regulatory T 세포(Treg)보다 keratin-14 (K-14)를 발현하는 진피의 끝부분에서 높게 발현되었다. Interlukin-10 (IL-10)는 Treg 뿐만 아니라 T helper 17 (Th17) 세포, dendritic 세포, macrophage 중 어느것과도 같은 위치에서 검출되지 않았다. 우리의 데이터는 방사선피부염 동물 모델의 병소 안에서, TGF-β1이 증식된 keratinocyte에 과발현된다는 것을 나타낸다. In this study, we established a radiodermatitis animal model and investigated the change in immune cell proportions in the secondary lymphoid organs. The cells responsible for the increased transforming growth factor-β1 (TGF-β1) and interleukin-10 (IL-10) production in the lesions following irradiation were also investigated. The radiodermatitis model was constructed by locally exposing the posterior dorsal region of hairless-1 (HR-1) mice to 10 Gy electron (E)-ray/day for six consecutive days. The change in immune cell proportions was analyzed by FACS. Immunohistochemistry was carried out to detect the expression of cytokines and cell-specific markers in the skin. The proportions of antigen-presenting cells, T cells, and B cells in the lymph nodes and spleen were affected by E-irradiation. After irradiation, TGF-β1 and IL-17 were co-localized in the papillary region of the dermis with keratin-14 (K-14)-positive cells rather than with regulatory T cells (Treg). IL-10 was not co-stained with Treg, T helper 17 (Th17) cells, dendritic cells, or macrophages. Our data indicate that TGF-β1 is over-expressed mainly by proliferated keratinocytes in the lesions of a radiodermatitis animal model.

Anti-obesity effect of Immature Persimmon (Diospyros kaki) Extract and its Fractions in 3T3-L1 cells

Seon-A Yoon,Boram Go,Ho Bong Hyun,Sung Chun Kim,Hyejin Hyeon,Dae-Ju Oh,Weon-Jong Yoon,Yong-Hwan Jung,Young-Min Ham 한국식품영양과학회 2020 한국식품영양과학회 학술대회발표집 Vol.2020 No.10