rDNA-ITS 염기서열 분석을 통한 시호 종 감별용 유전자 마커 개발 및 유연관계 분석

문병철 ( Byeong Cheol Moon ),추병길 ( Byeong Kil Choo ),지윤의 ( Yun I Ji ),윤태숙 ( Tae Sook Yoon ),이아영 ( A Young Lee ),전명숙 ( Myeong Sook Cheon ),김보배 ( Bo Bae Kim ),김호경 ( Ho Kyoung Kim ) 대한본초학회 2009 大韓本草學會誌 Vol.24 No.3

Objectives: Bupleuri Radix (Siho) is prescribed as the root of different Bupleurum species on the pharmarcopoeia in Korea and China. Moreover, other species and varieties of the genus Bupleurum have been also distributed on the herbal market as Bupleuri Radix. However, due to the morphological similarity and frequent occurrence of intermediate forms, the correct identification of this radix is very difficult. To develop a reliable method for correct identification and improving the quality standards of official Bupleuri Radix, we analyzed sequences of the ribosomal RNA gene and internal transcribed spacer (rDNA-ITS) region. Methods: PCR amplification of rDNA-ITS region was performed using ITS1 and ITS4 primer from 6 Bupleurum species and 1 variety, B. falcatum L. (Siho), an improved breed of B. falcatum L. (Samdo-Siho), B. chinense DC. (Buk-Siho), B. scorzonerifolium Willd. (Nam-Siho), B. longiadiatum Turcz. (Gae-Siho), B. euphorbiodes Nakai (Deungdae-Siho) and B. latissimum Nakai (Seom-Siho), and nucleotide sequence was determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW using entire rDNA-ITS sequence of three samples per species. Results: In comparative analysis of the rDNA-ITS sequences, we found specific nucleotides to distinguish Korean (B. falcatum L. and its variety) and Chinese official species (B. chinense DC. and B. scorzonerifolium Willd.) from others at positions 411 and 447, and positions 89, 101, 415 and 599, respectively. Futhermore, we also found nucleotide indels (insertion and/or deletion) and substitutions to identify each of different Bupleurum species, 2 positions for B. falcatum L. and its variety, 6 positions for B. chinense DC., 49 positions for B. scorzonerifolium Willd., 8 positions for B. euphorbioides Nakai, 7 positions for B. longiradiatum Nakai and 9 positions for B. latissimum Nakai. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origins of Bupleuri Radix. Moreover, we confirmed the phylogenetic relationship of B. latissimum Nakai, a Korean endemic speices, among Bupleurum species based on the rDNA-ITS sequence. Conclusions: These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterant Bupleurum species.

RAPD 분석을 통한 대황(大黃)과 종대황(種大黃) 감별용 SCAR 유전자 마커 개발

문병철 ( Byeong Cheol Moon ),이영미 ( Young Mi Lee ),천진미 ( Jin Mi Chun ),이아영 ( A Young Lee ),윤태숙 ( Taesook Yoon ),전명숙 ( Myeong Sook Cheon ),추병길 ( Byung Kil Choo ),김호경 ( Ho Kyoung Kim ) 대한본초학회 2009 大韓本草學會誌 Vol.24 No.4

Objectives: Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker. Methods: To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and mutiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively. Results: We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. Conclusions: These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.

matK 증폭용 primer 개발 및 염기서열 분석을 통한 정력자(葶藶子) 유전자 감별

문병철 ( Byeong Cheol Moon ),김욱진 ( Wook Jin Kim ),양선규 ( Sungyu Yang ),박인규 ( Inkyu Park ),여상민 ( Sang Min Yeo ),노푸름 ( Pureum Noh ) 대한본초학회 2018 大韓本草學會誌 Vol.33 No.3

Objectives : Lepidii seu Descurainiae Semen has been frequently adulterated with the seeds of several inauthentic plant species. However, the accurate identification of these plant seeds is very difficult. To develop a reliable genetic authentication tool for Lepidii seu Descurainiae Semen, we analyzed mat K sequence. Methods : To obtain the mat K sequences of plant materials, genomic DNA was extracted from 24 samples and PCR amplification was carried out using matK-AF/matK-8R universal primer set and matK-LDSF/matK-LDSR primer set. For identifying species-specific nucleotides and phylogenetic analysis, matK regions were sequenced and comparatively analyzed by the ClustalW and Maximum Likelihood method. Results : We developed a new primer set to amplify matK region in Lepidii seu Descurainiae Semen and closely related plant samples. From the comparative analysis of mat K sequences, we identified species-specific marker nucleotides for D. sophia, L. apetalum, L. latifolium, E. cheiranthoides, E. macilentum, and D. nemorosa, respectively. Furthermore, phylogenetic analysis revealed clear classification depending on the species. These results indicated that the matK sequence obtained a new primer set in this study was useful to identify Lepidii seu Descurainiae Semen in species level. Conclusions : We developed a primer set and identified species-specific marker nucleotides enough to distinguish authentic Lepidii seu Descurainiae Semen and adulterants at the species level based on the matK sequences. These genetic tool will be useful to prevent adulteration and to standardize the quality of Lepidii seu Descurainiae Semen.

DNA 바코드 분석을 통한 소계(小계) 및 대계(大계) 기원식물 감별과 종간 유연관계 분석

문병철 ( Byeong Cheol Moon ),이영미 ( Young Mi Lee ),지윤의 ( Yun Ui Ji ),최고야 ( Goya Choi ),천진미 ( Jin Mi Chun ),김호경 ( Ho Kyoung Kim ) 대한본초학회 2013 大韓本草學會誌 Vol.28 No.3

Objectives : The origin of Breeae Herba (So-gye) and Cirsii Herba (Dae-gye) is differently prescribed in Korean and Chinese modern pharmacopoeia. Since the similar morphological characteristics and chaotic plant names, moreover, the aerial part of Carduus crispus have been used as the Cirsii Herba. To develop a reliable method for correct identification of these herbal medicines and to evaluate the genetic relationship of these closely related plant species, we analyzed sequences of DNA barcode regions. Methods : Thirty-one samples of 6 medicinal plants (B. segeta, B. setosa, C. japonicum var. maackii, C. setidens, C. chanroenicum, and C. crispus) were collected from different habitate and nucleotide sequences of DNA barcode regions (rDNA-ITS, matK, and rbcL) were analyzed after amplification using appropriate primers reported in previous studies. The nucleotides of species-specific authentic marker and phylogenetic relations were estimated based on the entire sequences of DNA barcodes by the analysis of ClastalW and UPGMA, respectively. Results : In comparative analysis of DNA barcode sequences, we obtained specific nucleotides to discriminate the medicinal plant of Breeae/Cirsii Herba in species level and evaluated the phylogenetic relationship of these species. Futhermore, we identified distinct marker nucleotides enough to authenticate respective species. These sequence differences at corresponding positions were avaliable genetic markers to determine the botanical origins of Breeae Herbal as well as Cirsii Herba. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by providing of definitive information that can identify each plant species and distinguish from unauthentic adulterants and substitutes.

생물생산시설 및 환경공학 분야 : 돈사적용을 위한 공기순환식 태양열시스템의 효율분석

장진철 ( Jin Cheol Jang ),김명훈 ( Myeong Hun Kim ),문병은 ( Byeong Eun Moon ),김종구 ( Jong Ku Kim ),유영선 ( Young Sun Ryou ),김현태 ( Hyeon Tae Kim ) 한국농업기계학회 2014 한국농업기계학회 학술발표논문집 Vol.19 No.1

가축 생체 및 환경정보 분석을 통한 체중 추정 모델 개발

문병은 ( Byeong Eun Moon ),이민호 ( Min Ho Lee ),최태현 ( Tae Hyun Choi ),장진철 ( Jin Cheol Jang ),와카스카심 ( Waqas Qasim ),조재민 ( Jae Min Jo ),프랭크오취리 ( Frank Gyan Okyere ),김현태 ( Hyeon Tae Kim ) 한국농업기계학회 2017 한국농업기계학회 학술발표논문집 Vol.22 No.2

국내 축산업은 사육 규모의 증가 및 생산성 향상을 통해 산업적 측면에서 크게 성장하였다. 그러나 이러한 과정 속에서 사육시설의 무분별한 대형화, 밀집화는 오히려 구제역, AI 등 가축질병의 빈번한 발생 및 빠른 확산 등의 문제를 발생시킴에 따라 많은 경제적 피해가 발생하고 있다. 따라서 최근 사육시설의 대형화, 밀집화 문제와 질병 매개체로 인식되고 있는 축산 환경에 대한 개선 요구가 증가하고 있다. 이러한 국내 축산업의 문제점을 개선을 위해 최근 ICT, IoT 기반 가축 생체 및 환경정보 수집, 관리 시스템이 개발되어 현장에서 활용되고 있다. 그러나 현재까지는 주로 모니터링 및 자료의 저장, 수집 장치의 개선 및 제어에 국한되어 활용되고 있으며, 자동화 시스템을 위한 생체 및 환경정보 간 연관성 분석 및 모델 개발 연구는 거의 이루어지지 않았다. 돼지 사육 단계별 체중은 돼지의 건강 상태를 파악할 수 있는 가장 기본적인 물리적 생체정보로써 개체별 사료 급이량 및 급수량의 조절을 통한 사양관리의 개선과 온도 및 상대습도 등의 돈사 내부 환경 개선에 활용 가능하다. 따라서 본 연구에서는 돼지 체중의 추정 및 성장 모델 개발을 목표로 체중에 영향을 미치는 생체 및 환경정보 인자 간 연관성 분석을 실시하였다. 실험은 제주시에 위치한 D농장 비육사 1동 내 약 40~50 kg의 비육돈 50마리를 대상으로 하였다. 돼지등부의 면적을 산출하기 위해 급이 전 단계에서 체중 측정과 동시에 돼지의 머리에서 꼬리 방향으로 카메라가 이동하며 5회에 걸쳐 등부의 영상을 수집하였다. 촬영된 사진 중 돼지의 형상이 모두 측정된 사진을 필터링 후 이진화를 진행하였으며, 이진화된 영상을 기반으로 등부의 면적을 산출하였다. 실험기간 동안의 돈방의 환경정보를 측정하기 위해 가축환경모니터링시스템(LEMS)을 이용하였으며, 1분 단위로 온습도, CO₂, 풍속, 실효온도를 측정하여 등부의 면적과 환경정보 간의 연관성을 분석하였다.

가시박 군락지의 잡초발생특성 및 분포

문병철(Byeong Chul Moon),오세문(Se Mun Oh),이인용(In Yong Lee),김창석(Chang Seok Kim),조정래(Jeong Rae Cho),김석철(Seok Cheol Kim) 韓國雜草學會 2008 Weed&Turfgrass Science Vol.28 No.2

황납추출물이 도파민세포 보호효과 및 파킨슨병 행동장애에 미치는 영향

임혜선,문병철,박건혁,Lim, Hye-Sun,Moon, Byeong Cheol,Park, Gunhyuk 한국환경과학회 2022 한국환경과학회지 Vol.31 No.5

Parkinson's Disease (PD) is a chronic neurodegenerative disorder caused by the progressive loss of dopaminergic neurons, leading to decreased dopamine levels in the midbrain. Although the specific etiology of PD is not yet known, oxidative stress, inflammation, and subsequent apoptosis have been proposed to be closely related to PD pathophysiology. Cera Flava (CF) is a natural extract obtained from beehives and is isolated through the heating, compression, filtration, and purification of beehives. CF has been used in traditional medicines for its various clinical and pharmacological effects. However, its effects on neurodegenerative diseases are unknown. Therefore, we investigated the effects of CF against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD in mice and explored the underlying mechanism of action. In MPTP-induced PC12 cells, CF protected NADH dehydrogenase activity and inhibited lactate dehydrogenase. In the mouse model, CF promoted recovery from movement impairments, prevented dopamine depletion, and protected against MPTP-induced dopaminergic neuronal degradation. Moreover, CF downregulated glial and microglial activation. Taken together, our results suggest that CF improves behavioral impairments and protects against dopamine depletion in MPTP-induced toxicity by inhibiting glial and microglial activation.

포제에 따른 지황의 효능 변화에 관한 연구동향 분석

이지혜 ( Ji Hye Lee ),문병철 ( Byeong Cheol Moon ),최고야 ( Goya Choi ) 한약응용학회 2021 한약응용학회지 Vol.21 No.1

Processing is a traditional pharmaceutical technique in Korean medicine. Processing is mainly performed to enhance pharmacological effect of herbal medicine depending on specific clinical need. The root of Rehmannia glutinosa (Gaertn.) DC. (RR) is one of major herbal material in Korean medicine. Since nature of RR is altered by processing, the clinical use of RR varies based on processing method. Generally, fresh RR is used to treat blood-heat syndrome, while steamed RR is used to treat Yi-deficiency syndrome. This review summarized the up-to-date studies to investigate how processing affects pharmacological properties of RR. Using electronic database, a total of 13 studies were collected and included in this study. These studies exhibited pharmacological properties of RR including immunostimulatory, hematopoietic, blood-cooling, anti-oxidant, anti-tumor, anti-thrombosis and anti-ulcer effects. All of studies were conducted by using in vitro or in vivo bioassays. This study could help to establish the basis for future research on processing of RR. In order to provide comprehensive data, further study is needed which includes changes in chemical profiles and analysis of relation between chemical profiles changes and pharmacological properties.

피부각질형성세포에서 동의보감 충부약재 14 종이 염증성 사이토카인 생성에 미치는 영향

박건혁 ( Gunhyuk Park ),문병철 ( Byeong Cheol Moon ),임혜선 ( Hye-sun Lim ) 대한화장품학회 2020 대한화장품학회지 Vol.46 No.2

본 연구에서는 피부각질형성세포에서 동의보감에 기록된 14 종의 충부약재에 대한 염증성 사이토카인 생성에 미치는 영향을 조사하였다. 이를 확인하기 위하여 멀티플렉스 사이토카인 분석으로 17 종에 대한 사이토카인을 스크리닝 하였다. 충부약재 14 종 중, 제조 및 전갈 추출물은 IL-5 생성을 억제하였고, 제조, 상표초 및 수질 추출물은 IL-6 생성을, 제조, 상표초, 지룡, 수질 및 문합 추출물은 IL-8 생성에 억제 효과를 보였다. 합개, 잠사, 선퇴 및 문합 추출물은 IL-13 생성을 억제하였으며, 귀판, 잠사 및 지룡 추출물은 MIP-1β 생성에 유의적인 효과를 확인하였다. 이러한 충부약재는 피부염증 억제를 위한 항염증제 활용 가능성에 의미를 둘 수 있지만, 그 가치를 증명하기 위해서는 피부개선을 위한 기전연구를 포함하여 인체피부모사판 모델에서 추가 실험을 통한 검증이 선행 되어야 한다. The purpose of this study was to investigate the effects of 14 Chung-bu medicinal materials described in the Dongui Bogam on inflammatory cytokine production in HaCaT human keratinocyte cells. In order to confirm this possibility, we screened inhibition activity of 17 cytokines using Bio-Plex Pro™ Human Cytokine 17-plex assay in HaCaT cell lines. Of the 14 Chung-bu medicinal materials, Holotrichia (Ho) and Scorpio (Sc) exerted inhibitory effects on interleukin (IL)-5 production; Ho, Mantidis Ootheca (MO), and Hirudo (Hi) exerted inhibitory effects on IL-6 production; Ho, MO, Lumbricus (Lu), Hi, and Meretricis Concha (MC) showed significant inhibitory effects on IL-8 production; Gecko (Ge), Bombycis Faeces (BF), Cicadidae Periostracum (CP), and MC showed significant inhibitory effects on IL-13 production; and Testudinis Chinemis Plastrum et Carapax (TCPC), BF, and Lu exerted significant inhibitory effects on MIP-1β production. Results indicated that the Chung-bu medicinal materials might be a good candicate as potential anti-inflammatory agents for inhibition of skin inflammation. However, further investigations on these materials, including mechanistic studies, should be carried out to validated the effects in human skin equivalent models of dermatitis.