발효 / 효소 분야

지승택,신현길 한국식품과학회 1999 한국식품과학회 학술대회 Vol.1999 No.1

Sprague - Dawley - Rat 의 Primary 대장세포에 있어서 유산균들의 In Vitro 항유전 독성효과

지승택,박종흠,최옥병,신현길,현창기 한국축산식품학회 1999 한국축산식품학회지 Vol.19 No.1

작약(Paeoniae radix) 추출물의 식후 과혈당 억제작용

지승택(Seung-Taek Ji),이성진(Sung-Jin Lee),이강은(Kang-Eun Lee),손용태(Yong-Tae Son),정요경(Yo-Kyung Chung) 한국식품영양과학회 2002 한국식품영양과학회지 Vol.31 No.1

본 연구에서는 작약 추출물의 식후 과혈당 억제작용을 조사하였다. 작약 유기용매(hexane, ethyl acetate, butanol, water)추출물들을 high performance liquid chromatography를 이용하여 분획한 분획물(fraction)들의 α-glucosidase(EC 3. 2. 1. 20) 저해제를 탐색하였다. 작약 ethyl acetate 추출물의 11, 12, 18, 19번 분획물들은 20 μg/mL에서 각각 85%, 84%, 77%, 77%의 강력한 저해 활성을 나타내었으며, 작약 ethyl acetate 추출물의 10~19번 분획을 경구투여한 in vivo당(maltose) 부하 실험에서도 양성대조군과 비교하여 유의성 있게 22%의 혈당을 낮추었다. 따라서 본 연구의 in vitro와 in vivo 실험을 통하여 확인 된 α-glucosidase 저해 활성을 갖는 작약 분획물들은 식후의 혈당상승 작용을 억제하는 새로운 nutraceutical 소재와 항당뇨 신약개발을 위한 탐색자원으로서 매우 가치있는 자원으로 기대된다. This study was carried out to investigate the inhibitory effect of extracts from Paeoniae radix on postprandial hyperglycemia. Organic solvent (hexane, ethyl acetate, butanol, water) extracts from Paeoniae radix were fractionated by high performance liquid chromatography. These fractions were used to screen α-glucosidase (EC 3. 2. 1. 20) inhibitors by microplate colorimetric assay. The fractions 11, 12, 18, 19 of ethyl acetate extract from Paeoniae radix showed inhibitory activity by 85%, 84%, 77%, 77% at concentration of 20 μg/mL, respectively. The selected fractions (no. 10~no. 19) significantly reduced by 22% the blood glucose elevation in comparison with positive control in mice loaded with maltose. The fractions of Paeoniae radix were determined in vitro inhibitory activity on α-glucosidase and in vivo inhibition effect on blood glucose elevation in mice. Therefore, these results suggest that the extract of Paeoniae radix can be used as a new nutraceutial for inhibition on postprandial hyperglycemia as well as resource pool for lead compounds as a α-glucosidase inhibitor.

Comet Assay를 이용한 전통발효식품인 배추김치의 항유전 독성효과

지승택(Seung-Taek Ji),박종흠(Jong-Heum Park),현창기(Chang-Kee Hyun),신현길(Heuyn-Kil Shin) 한국식품영양과학회 2000 한국식품영양과학회지 Vol.29 No.2

본 연구에서는 우리나라 전통발효식품인 배추김치의 암예방 기능성을 규명하기 위하여 항유전독성을 측정할수 있는 Comet assay(일명 single cell microgel electrophoresis)를 사용하였다. 배추김치를 물 분획, 유기용매(n-hexane, chloroform, ethyl acetate) 분획, 불용성 분획과 우세발효균주들로 분리하였다. 배추에서 분리되어진 물 분획, n-hexane 분획, chloroform 분획, ethyl acetate분획, 불용성 분획은 non-tumoral normal 3T3 세포의 DNA손상에 대하여 항유전 독성효과를 나타내지 못하였다. 그러나 배추에서 분리된 발효균주 bacteria 1과 bacteria 2는 유의성 있게 3T3 세포의 DNA 손상을 감소시키는 항유 전독성 효과를 나타내었으나 bacteria 3, 4, 5는 항유전독성 활성을 나타내지 않았다. This study carried out to elucidate the cancer chemoprevention of Korean native fermented food, baechu kimchi using Comet assay (in other words, single cell microgel electrophoresis). For this purpose, baechu kimchi was fractionated by water, n-hexane, chloroform and ethyl acetate. 5 strains of dominant fermented bacteria were isolated from baechu kimchi. The water fraction, n-hexane fraction, chloroform fraction, ethyl acetate fraction and water insoluble fraction showed no antigenotoxicities in non-tumoral normal 3T3 cells. Among 5 bacteria isolates from baechu kimchi, two isolates bacteria 1 and 2 strongly inhibited genotoxicity of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in non-tumoral normal 3T3 cells (p<0.05). Bacteria 3, 4 and 5 were also not antigenotoxic.

Long-Term Priming by Three Small Molecules Is a Promising Strategy for Enhancing Late Endothelial Progenitor Cell Bioactivities

김연주,지승택,김다연,정석윤,강송화,박지혜,장웅비,윤지수,하종성,이동형,권상모 한국분자세포생물학회 2018 Molecules and cells Vol.41 No.6

Endothelial progenitor cells (EPCs) and outgrowth endothelial cells (OECs) play a pivotal role in vascular regeneration in ischemic tissues; however, their therapeutic application in clinical settings is limited due to the low quality and quantity of patient-derived circulating EPCs. To solve this problem, we evaluated whether three priming small molecules (tauroursodeoxycholic acid, fucoidan, and oleuropein) could enhance the angiogenic potential of EPCs. Such enhancement would promote the cellular bioactivities and help to develop functionally improved EPC therapeutics for ischemic diseases by accelerating the priming effect of the defined physiologi-cal molecules. We found that preconditioning of each of the three small molecules significantly induced the differentiation potential of CD34+ stem cells into EPC lineage cells. Notably, long-term priming of OECs with the three chemical cocktail (OEC-3C) increased the pro-liferation potential of EPCs via ERK activation. The migration, invasion, and tube-forming capacities were also significantly enhanced in OEC-3Cs compared with unprimed OECs. Further, the cell survival ratio was dra-matically increased in OEC-3Cs against H2O2-induced oxidative stress via the augmented expression of Bcl-2, a pro-survival protein. In conclusion, we identified three small molecules for enhancing the bioactivities of ex vivo-expanded OECs for vascular repair. Long-term 3C priming might be a promising methodology for EPC-based therapy against ischemic diseases.

Engineered M13 Peptide Carrier Promotes Angiogenic Potential of Patient-Derived Human Cardiac Progenitor Cells and In Vivo Engraftment

장웅비,지승택,박지혜,Kim Yeon-Ju,Kang Songhwa,김다연,이나경,김진수,Lim Hye Ji,최재우,LE THI HONG VAN,LY THANH TRUONG GIANG,비누스,김동환,하종성,윤지수,Baek Sang Hong,권상모 한국조직공학과 재생의학회 2020 조직공학과 재생의학 Vol.17 No.3

BACKGROUND: Despite promising advances in stem cell-based therapy, the treatment of ischemic cardiovascular diseases remains a big challenge due to both the insufficient in vivo viability of transplanted cells and poor angiogenic potential of stem cells. The goal of this study was to develop therapeutic human cardiac progenitor cells (hCPCs) for ischemic cardiovascular diseases with a novel M13 peptide carrier. METHOD: In this study, an engineered M13 peptide carrier was successfully generated using a QuikChange Kit. The cellular function of M13 peptide carrier-treated hCPCs was assessed using a tube formation assay and scratch wound healing assay. The in vivo engraftment and cell survival bioactivities of transplanted cells were demonstrated by immunohistochemistry after hCPC transplantation into a myocardial infarction animal model. RESULTS: The engineered M13RGD?SDKP peptide carrier, which expressed RGD peptide on PIII site and SDKP peptide on PVIII site, did not affect morphologic change and proliferation ability in hCPCs. In contrast, hCPCs treated with M13RGD?SDKP showed enhanced angiogenic capacity, including tube formation and migration capacity. Moreover, transplanted hCPCs with M13RGD?SDKP were engrafted into the ischemic region and promoted in vivo cell survival. CONCLUSION: Our present data provides a promising protocol for CPC-based cell therapy via short-term cell priming of hCPCs with engineered M13RGD?SDKP before cell transplantation for treatment of cardiovascular disease.

Engineered M13 Nanofiber Accelerates Ischemic Neovascularization by Enhancing Endothelial Progenitor Cells

이준희,김성욱,지승택,김연주,장웅비,오진우,김재호,유소영,백상홍,권상모 한국조직공학과 재생의학회 2017 조직공학과 재생의학 Vol.14 No.6

Dysfunction or loss of blood vessel causes several ischemic diseases.Although endothelial progenitor cells (EPCs) are a promising source for cell-based therapy, ischemia-induced pathophysiological condition limits the recovery rate by causing drastic cell death. To overcome this issue, we attempted to develop a cell-targeted peptide delivery and priming system to enhance EPCbased neovascularization using an engineered M13 bacteriophage harboring nanofibrous tubes displaying *2700 multiple functional motifs. The M13 nanofiber was modified by displaying RGD, which is an integrin-docking peptide, on the minor coat protein, and bymutilayering SDKPmotifs,which are the key active sites for thymosin b4, on themajor coat protein. The engineered M13 nanofiber dramatically enhanced ischemic neovascularization by activating intracellular and extracellular processes such as proliferation, migration, and tube formation in the EPCs. Furthermore, transplantation of the primed EPCs with the M13 nanofiber harboring RGD and SDKP facilitated functional recovery and neovascularization in a murine hindlimb ischemia model. Overall, this study demonstrates the effectiveness of theM13 nanofiber-based novel peptide deliveryandprimingstrategy inpromotingEPC bioactivity and neovessel regeneration. To our knowledge, this is first report onM13 nanofibers harboring dual functional motifs, the use of which might be a novel strategy for stem and progenitor cell therapy against cardiovascular ischemic diseases.

Comet assay를 이용한 방사선 조사육의 판별

정석규,박종흠,지승택,박금주,김해홍,현창기,신현길,Jeong, Seok-Kyu,Park, Jong-Heum,Ji, Seung-Taek,Park, Kum-Ju,Kim, Hai-Hong,Hyun, Chang-Kee,Shin, Heuyn-Kil 한국식품과학회 2000 한국식품과학회지 Vol.32 No.4

Comet assay를 이용하여 방사선 조사육의 조사여부 및 조사량을 판별해내는 방법을 개발하기 위해 1-10 kGy의 감마선 조사량으로 조사한 육조직에서 일어나는 DNA 손상을 측정하였다. Comet assay에 있어 최적의 comet image를 얻기 위해 세포 분리, 세포 lysis 및 전기영동에 대한 여러 조건들을 적용하여 최적의 조건을 확립하였다. DNA 손상도는 관찰되는 comet의 평균 tail length와 tail length에 의해 구분된 4 손상 등급에의 분포, 그리고 그 분포비율에 의해 본 연구에서 제시한 식에 의해 계산한 relative damage index(RDI) 값 등으로 비교 판정하였다. 평균 tail length와 RDI 값은 조사량이 증가함에 따라 증가하여 DNA 손상도가 증가됨을 나타내었으며, 평균 tail length으로는 조사량 간의 차이를 명확히 분별하기가 어려웠던 반면 RDI 값에 의하면 조사여부 및 조사량을 판별하기에 적합함을 알 수 있었다. 국내산 한우육과 수입 냉장육에 대하여 blind test를 실시한 결과 수입육이 높은 DNA 손상도를 나타내었는데 그 RDI 값은 방사선 조사에 의한 값보다는 비교적 낮은 것이어서 수입육의 DNA 손상은 방사선 조사가 아닌 저온처리의 결과인 것으로 판단되었다. 본 연구의 결과로부터 Comet assay가 우육의 방사선 조사여부 및 조사량 판별에 유용한 기술로 응용될 수 있을 것으로 판단되었다. DNA damages in post-mortem bovine muscle samples caused by gamma irradiation at doses of 1 to 10 kGy were determined by Comet assay. When the cell extract was prepared in a physical method and followed by neutral lysis and neutral electrophoresis, the optimal comet images could be obtained. DNA damages were evaluated from the mean tail length, the distributions of comet images in 4 groups divided by tail length and the relative damage index (RDI) values calculated from the distribution pattern. The mean tail length and RDI value were increased by increasing the irradiation dose, and the RDI value was found to be useful as an index for discriminating of irradiation and measuring the irradiated dose. Blind tests using Korean domestic (Hanwoo) and imported beef samples showed a higher RDI value for the latter. However, the value was lower than those of irradiated samples indicating that the cause of DNA damages in the imported beef samples might be not irradiation but low-temperature treatments. It was concluded from the results of this study that the irradiated beef and its irradiated dose could be detected and predicted by Comet assay.

황련 ( Coptis chinensia Franch ) 의 추출물로부터 포자발아 억제물질의 분리 및 동정과 그 효과

신현길,김종배,최원희,지승택,최경숙 한국축산식품학회 1994 한국축산식품학회지 Vol.14 No.1

Lnk is an important modulator of insulin-like growth factor-1/ Akt/peroxisome proliferator-activated receptor-gamma axis during adipogenesis of mesenchymal stem cells

이준희,이상훈,이향선,지승택,정석윤,김재호,배순식,권상모 대한약리학회 2016 The Korean Journal of Physiology & Pharmacology Vol.20 No.5

Adipogenic differentiation of mesenchymal stem cells (MSCs) is critical for metabolic homeostasis and nutrient signaling during development. However, limited information is available on the pivotal modulators of adipogenic differentiation of MSCs. Adaptor protein Lnk (Src homology 2B3 [SH2B3]), which belongs to a family of SH2-containing proteins, modulates the bioactivities of different stem cells, including hematopoietic stem cells and endothelial progenitor cells. In this study, we investigated whether an interaction between insulin-like growth factor-1 receptor (IGF-1R) and Lnk regulated IGF-1-induced adipogenic differentiation of MSCs. We found that wild-type MSCs showed greater adipogenic differentiation potential than Lnk–/– MSCs. An ex vivo adipogenic differentiation assay showed that Lnk–/– MSCs had decreased adipogenic differentiation potential compared with wild-type MSCs. Interestingly, we found that Lnk formed a complex with IGF-1R and that IGF-1 induced the dissociation of this complex. In addition, we observed that IGF-1-induced increase in the phosphorylation of Akt and mammalian target of rapamycin was triggered by the dissociation of the IGF-1R–Lnk complex. Expression levels of a pivotal transcription factor peroxisome proliferatoractivated receptor gamma (PPAR-γ) and its adipogenic target genes (LPL and FABP4) significantly decreased in Lnk –/– MSCs. These results suggested that Lnk adaptor protein regulated the adipogenesis of MSCs through the IGF-1/Akt/PPAR-γ pathway.