넓패(Ishige foliacea)의 최종당화산물 생성저해 활성 및 신장세포 보호 효과

조치흥 ( Chi Heung Cho ),김민경 ( Mingyeong Kim ),염가희 ( Ga Hee Youm ),김세라 ( Sera Kim ),박용곤 ( Yongkon Park ),이상훈 ( Sang-hoon Lee ) 한국키틴키토산학회 2020 한국키틴키토산학회지 Vol.25 No.3

본 연구에서 국내 자생 갈조류 중 하나인 넓패를 이용하여 추출방법에 따른 생리활성 물질의 함량을 정량적으로 분석하였다. 또한 당뇨병성신장증의 합병증을 유발하는 물질로 알려진 최종당화산물의 생성 억제 효과, 최종당화산물과 콜라겐 사이의 교차결합 형성 억제 효과 및 교차결합 절단 효과를 평가하였다. 넓패 70% 에탄올 추출물은 총 페놀, 총 플라보노이드 및 탄닌과 같은 생리활성 성분을 가장 많이 함유하는 추출 조건임을 확인하였으며, 최종당화산물의 생성 억제, 최종당화산물과 콜라겐 사이의 가교결합 형성 저해 및 생성된 가교결합을 절단하는 효과 또한 가장 높았다. 특히, 70% EtOH IFE는 양성대조군으로 처리한 AG보다 더 높은 교차 결합 억제 및 교차결합 절단 효과를 나타내는 것을 확인할 수 있었다. 또한 70% EtOH IFE는 MGO에 의해 유도되는 산화적 스트레스로부터 신장 세포인 mesangial 세포의 세포 내 산화적 스트레스 수준을 유의적으로 감소시킬 뿐만 아니라 apoptosis로 인한 세포 사멸을 감소시켜 신장 손상을 보호하는 효과를 보였다. 따라서 넓패는 부작용이 적은 당뇨합병증 예방 및 치료제 개발 후보물질로서 가능성이 충분한 것으로 판단되며, 향후 국내 자생 해양자원의 가치 향상을 위해 넓패의 생리활성 성분의 동정 및 당뇨합병증 예방과 치료에 작용하는 메커니즘 구명 연구가 추가적으로 진행되어야 할 것으로 사료된다. The present study was conducted to investigate the anti-glycation activities and renoprotective properties of Ishige foliacea. The yield of extract ranged from 3.20 to 35.59%. Among the extracts, 70% aqueous ethanol extract of Ishige foliacea (70% EtOH IFE) showed the highest total phenolic, condensed tannin, and total flavonoid contents (1.570 ± 0.165 mg GAE/g DW, 0.407 ± 0.008 mg CE/g DE, 0.353 ± 0.050 mg CE/g DE, respectively). 70% EtOH IFE showed the highest breaking activity against preformed AGEs-BSA cross-link than ALT-711, the well-known cross-link breaker. In addition, 70% EtOH IFE observedthe highest inhibitory effect on the AGEs-BSA cross-link and exhibited more potent than aminoguanidine, a well-known AGEs-BSA cross-link inhibitor. Treatment with the 70% EtOH IFE, at 2, 10, and 50 μg/mL, reduced oxidative stress to the level of approximately 186.3-95.9%. The intracellular antioxidative effect of the 70% EtOH IFE was stronger to that with amino-guanidine at 500 μM. Pre-treatments of cells with the 70% EtOH IFE at concentrations of 2, 10, and 50 μg/mL led to 60.3%, 38.0%, and 20.8% decrease in apoptotic cells, respectively. These result suggest that 70% aqueous ethanol extract of Ishige foliacea can serve as a good source of natural anti-diabetic complications agents via breaking and inhibition of AGEs-BSA cross-link and renoprotective effects against MGO-induced oxidative stress.

우뭇가사리(Gelidium amansii)추출물의 Mouse Glomerular Mesangial Cells에최종당화산물로 유도된 당뇨병성 신병증 완화 효과

조치흥 ( Chi Heung Cho ),김민경 ( Mingyeong Kim ),김세라 ( Sera Kim ),이상훈 ( Sang-hoon Lee ) 한국키틴키토산학회 2021 한국키틴키토산학회지 Vol.26 No.4

본 연구에서는 제주도에서 자생하는 홍조류인 우뭇가사리 (Gelidium amansii) 70% (v/v) 에탄올 추출물(GAE)의 항당화 (anti-glycation) 효능 및 최종당화산물로 유도되는 당뇨병성 합병증 중 하나인 신장병증(nephropathy)에 대한 예방 효과를 평가하였다. GAE는 모든 처리 농도(1, 5, 및 20 μg/mL)에서 BSA, glucose, 및 fructose로 인해 생성되는 최종당화산물을 유의적으로 감소시키는 것을 확인하였다. 또한 GAE의 전처리는 최종당화산물과 콜라겐 단백질 사이의 교차결합 형성을 유의적으로 억제 시켰으며, 최종당화산물과 콜라겐 단백질 사이에 이미 형성된 비가역적 교차결합에 GAE를 처리하였을 때 매우 효과적으로 절단하는 효능을 가지고 있음을 확인하였다. GAE의 신장 보호 효과를 평가하기 위해 mouse glomerular mesangial 세포 보호 효능을 평가한 결과 최종당화산물의 전구체인 MGO로 유도된 세포 독성으로부터 신장 세포를 보호하는 효과를 나타내었다. 이러한 효과는 GAE의 전처리가 MGO로 인해 증가하는 신장 세포 내 ROS 생성 억제, 세포 내 최종당화산물 축적 감소, 및 세포 내 MGO 축적 감소를 통해 신장 세포의 apoptosis를 억제하기 때문인 것을 확인하였다. 본 연구를 통해 GAE는 합성 당뇨합병증 치료제로 알려진 아 미노구아니딘과 유사한 수준의 효능을 가지고 있음을 확인하였다. 그러므로 우뭇가사리는 최종당화산물로 인해 유도되는 당뇨병성 신병증을 예방 및 치료하는 효과를 가지고 있는 것으로 보이며, 부작용 위험이 낮은 천연 당뇨합병증 치료제로 개발 잠재력이 높은 것으로 사료된다. In this study, we evaluated the anti-glycation activity and nephroprotective effects of Gelidium amansii 70% (v/v) ethanol extract (GAE) in mouse glomerular mesangial cells. GAE (1, 5, and 20 μg/mL) treatment significantly reduced advanced glycation end-products (AGEs) formation (p < 0.001), and inhibited AGEs formation up to 47.8% at 1 μg/mL GAE. In addition, GAE not only significantly inhibited the formation of AGEs-collagen cross-linking, but also showed the effect of breaking the already formed cross-links (p < 0.001). In particular, 20 μg/mL GAE (41.9%) exhibited similar breaking ability as ALT-711 (0.5 mg/mL, 33.7%) used as a positive control. Treatment of methylglyoxal (MGO), a precursor of AGEs, on mouse glomerular mesangial cells, increased intracellular MGO and AGEs accumulation, leading to increased reactive oxygen species (ROS) production and eventually induced renal cell apoptotic death. However, pre-treatment with the GAE, at 1, 5 and 20 μg/mL, reduced intracellular ROS production level to approximately 541.1-201.4%. Especially, pre-treatment of 20 μg/mL GAE significantly decreased intracellular MGO concentration (18.99 μg/mL) to a level similar to that of the non-treated group (16.67 μg/mL) (p < 0.001). Moreover, it was confirmed by immunofluorescence analysis that GAE suppressed the AGEs accumulation in mouse glomerular mesangial cells. As a result, due to this effects of GAE, MGO-induced apoptotic cell death was dramatically reduced, proving that GAE has an ability in preventing AGEs-related diabetic nephropathy. Therefore, our results suggest that Gelidium amansii has high potential to be developed as a natural agent for diabetic complications with a low risk of side effects.

Optimization of Recombinant Human Platelet-Derived Growth Factor-BB Encapsulated in Poly (Lactic-co-Glycolic Acid) Microspheres for Applications in Wound Healing

배윤주,조치흥,이우종,허증수,임정옥 한국조직공학과 재생의학회 2016 조직공학과 재생의학 Vol.13 No.1

Growth factors play multiple and critical roles in wound repair processes. Platelet-derived growth factor (PDGF) is a potent growth factor that is particularly important in the early inflammatory phase of wound healing. In order to extend the half-life of PDGF, polymeric encapsulation is used. In the current study, Poly (lactic-co-glycolic acid) (PLGA) microspheres containing recombinant human (rh) PDGF-BB were prepared to prolong the effectiveness of this growth factor. PLGA microspheres were optimized using a modified w/o/w-double-emulsion/solvent evaporation method by changing the processing conditions of stirring speed and emulsifier (polyvinyl alcohol) concentration. Microspheres prepared using the optimized method released rhPDGF-BB for up to three weeks. An in vitro migration assay showed a significant decrease in the wound area in cells treated with rhPDGF-BB microspheres compared to control cells. These findings demonstrate the potential of rhPDGF-BB encapsulated in microspheres to enhance wound healing.

Kiwifruit of Actinidia eriantha cv. Bidan has in vitro antioxidative, anti-inflammatory and immunomodulatory effects on macrophages and splenocytes isolated from male BALB/c mice

김영은,조치흥,강희,허호진,조윤섭,김대옥 한국식품과학회 2018 Food Science and Biotechnology Vol.27 No.5

Kiwifruit is known to contain considerable amount of antioxidative phenolics. The objective of this study was to evaluate the antioxidative, anti-inflammatory and immunomodulatory effects of Actinidia eriantha cv. Bidan and A. deliciosa cv. Hayward kiwifruits. The antioxidant capacity of kiwifruit was measured with the DPPH, ABTS and ORAC assays, and was significantly (p\0.05) higher in cv. Bidan than in cv. Hayward. The production of proinflammatory cytokines interleukin-6, interleukin-12 and tumor necrosis factor-a by peritoneal macrophages from male BALB/c mice was significantly (p\0.05) lower following treatment of cv. Bidan extracts than after treatment with lipopolysaccharide alone. Cv. Bidan extracts significantly (p\0.05) increased the proliferation of splenocytes stimulated with an anti-CD3 antibody and significantly (p\0.05) reduced their interferon- c secretion. Taken together, these findings suggest that cv. Bidan kiwifruit is rich in antioxidants and may be a source of anti-inflammatory and immunomodulatory agents.

Effects of freeze-drying on antioxidant and anticholinesterase activities in various cultivars of kiwifruit (Actinidia spp.)

황정승,조치흥,백무열,박승국,허호진,조윤섭,김대옥 한국식품과학회 2017 Food Science and Biotechnology Vol.26 No.1

Oxidative stress contributes to neurodegenerative disorders such as Alzheimer’s disease. Phenolic antioxidants can efficiently reduce oxidative stress. In this study, we evaluated the effects of the freeze-drying process on phenolics, antioxidants, and cholinesterase inhibition in five cultivars of kiwifruits grown in Korea, Actinidia chinensis cv. Hort16A, cv. Happygold, and cv. Haegeum; A. deliciosa cv. Hayward; and A. eriantha cv. Bidan, by comparing them with their fresh counterparts. Among the five cultivars of both fresh and freeze-dried kiwifruits tested in this study, cv. Bidan had the highest levels of total phenolics, total flavonoids, and antioxidants, and cv. Hayward had the lowest. Freezedried kiwifruits inhibited acetylcholinesterase and butyrylcholinesterase that catalyze the breakdown of acetylcholine (neurotransmitter). On sensory evaluation, cv. Happygold had the highest overall preference scores among the freeze-dried kiwifruits. The results suggest that freeze-dried kiwifruit could serve as a good source of antioxidants and cholinesterase inhibitors.

Acetylated distarch adipate의 인간 장 상피세포 염증완화에 의한 위장관 투과성 개선 효능

Lee Sang-Hoon,김민경,조치흥,Kim Sera,오미진,Park Ho-Young 한국키틴키토산학회 2022 한국키틴키토산학회지 Vol.27 No.2

The intestine digests the food we eat and regulates the absorption of nutrients as well protect the body from external antigen or toxic substances. This barrier function become abnormal during inflammation. In this study, we confirmed the anti-inflammatory and epithelial barrier protective properties of acetylated distarch adipate (ADSP) in human-derived intestinal epithelial cells. ADSP showed a significant increase in cell viability with pro-inflammatory cytokine treatment and non-treatment. IL-6 and IL-1β significantly decreased with ADSP treatment as compared to the inflammation-induced group, while IL-8 didn’t show significant change. After confirming that ADSP significantly reduces the epithelial monolayer integrity damage and increased permeability induced by TNF-α and IFN-γ, we confirmed the change in expression amount of tight junction with immunofluorescence and western blotting. Our results contributed to the study of the bioactivity of ADSP, which is widely used as a food additive, and indicated its potential as an agent for inflammatory bowel disease.

청도반시(Diospyros kaki Thunb. cv. Cheongdo-Bansi) 탈삽 껍질 추출물의 산화스트레스로부터 PC-12 신경세포 보호 효과

정다울(Da-Wool Jeong),조치흥(Chi Heung Cho),라찬수(Chan Su Rha),이승환(Seung Hwan Lee),김대옥(Dae-Ok Kim) 한국식품과학회 2017 한국식품과학회지 Vol.49 No.5

본 연구에서는 떫은 감인 청도반시(Diospyros kaki Thunb. cv. Cheongdo-Bansi)를 탈삽하여 곶감으로 만드는 과정 중에 다량의 부산물로 생겨나는 감 껍질을 활용하고자 산화방지능 및 신경세포 보호능을 평가하였다. 탈삽한 감 껍질을 40% (v/v) 에탄올-물혼합용액을 사용하여 초음파 추출을 한 후에, 가열처리와 비가열처리 방법을 이용하여 각각의 추출물을 확보하였다. 가열처리한 추출물이 비가열처리로 얻어진 것에 비해서 총페놀 함량, 총플라보노이드 함량, 산화방지능에서 약 1.3-1.8배 더 높았다. 비가열처리 추출물과 비교하여, 가열처리한 것이 PC-12 신경세포의 생존율 더 높이고, 세포 내 산화스트레스를 좀 더 완화하였다. AChE와 BChE 억제능 역시 가열처리한 추출물이 비가열처리로 얻은 추출물보다 더 높았다. 이러한 결과들은 산화방지능을 보유하며 콜린가수분해효소를 억제하는 기능성 소재 발굴을 위한 탈삽 감껍질 추출물 활용 가능성을 보여준다. Astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) peel with the astringency removed, which is a by-product of dried persimmon (gotgam), was investigated for its antioxidant and neuroprotective properties. A mixture of peel and 40% (v/v) aqueous ethanol was subjected to ultrasonication and then thermal and nonthermal treatments, to produce thermally-treated and nonthermally-treated persimmon peel extracts (TPE and NTPE, respectively). The total phenolic and flavonoid contents and the antioxidant capacity of TPE was approximately 1.3-1.8 times higher than those of NTPE. TPE resulted in the increased viability of neuronal PC-12 cells compared with NTPE. Furthermore, intracellular oxidative stress in PC-12 cells was more decreased by treatment with TPE than NTPE. Cholinesterases, such as acetylcholinesterase and butyrylcholinesterase, were more inhibited by treatment with TPE than NTPE. These results suggest that TPE is useful as a functional material to decrease oxidative stress in neuronal cells and to inhibit cholinesterases.

불레기말(Colpomenia sinuosa)의 최종당화산물 저해 효능 및 신장 보호 효과

김민경(Mingyeong Kim),조치흥(Chi Heung Cho),김세라(Sera Kim),최인욱(In-Wook Choi),이상훈(Sang-Hoon Lee) 한국해양바이오학회 2021 한국해양바이오학회지 Vol.13 No.2

Here, we evaluated the anti-glycation effects and renal protective properties of 70% (v/v) ethanolic extract of Colpomenia sinuosa (CSE) against AGEs -induced oxidative stress and apoptosis at different concentrations (1, 5, and 20 μg/mL). At 20 μg/mL, CSE showed that antiglycation activities via the inhibition of AGE formation (51.1%), inhibition of AGEs–protein cross-linking (61.7%), and breaking of AGEs–protein cross-links (33.3%), were significantly (###p< 0.001 vs. non-treated group) lower than the nontreated group. Methylglyoxal (MGO) significantly (***p < 0.001) reduced cell viability (24.4%) and increased reactive oxygen species (ROS) level (642.3%), MGO accumulation (119.4 μg/mL), and apoptosis (55.0%) in mesangial cells compared to the nontreated group. Pretreatment with CSE significantly (###p < 0.001) increased cell viability (57.8%) and decreased intracellular ROS (96.5%), MGO accumulation (80.0 μg/mL), and apoptosis (22.6%) at 20 μg/mL. Additionally, we confirmed intracellular AGEs reduction by CSE pretreatment. Consequently, our results suggest that CSE is a good source of natural therapeutics for managing diabetic complications by the antiglycation effect and renal protective activity against MGO-induced oxidative stress.

열풍 건조 가공 공정이 다래 과실(품종명 만수)의 산화방지능에 미치는 영향

김성일(Cheng-Ri Jin),조치흥(Chi Heung Cho),남태규(Tae-Gyu Nam),조윤섭(Youn-Sup Cho),김대옥(Dae-Ok Kim) 한국식품과학회 2015 한국식품과학회지 Vol.47 No.4

다래 과실(품종명 만수)의 건조 절편을 제조하고자 열풍건조기를 이용하여 35, 50 및 70℃의 여러 온도에서 생과를 24시간 동안 열풍건조 하였다. 열풍건조 다래의 총 페놀 함량, 총 플라보노이드 함량과 산화방지능은 열처리하지 않은 생과(대조구)보다 감소하였다. ABTS 및 DPPH 라디칼 소거능에 기반하여 측정한 산화방지능과 총 페놀 함량 및 총 플라보노이드 함량과의 상관관계는 r²이 0.9 이상으로 높은 반면에, ORAC 법을 이용한 산화방지능은 상대적으로 낮은 상관관계를 나타내었다. 이러한 실험결과를 바탕으로 건조다래의 제조 시 온도 등 최적 가공 조건을 확립하여, 기능성이 우수한 건조다래를 개발할 필요가 있다. 또한, 고분자 함량이 많은 과피와 함께 과육의 섭취가 가능한 다래를 활용하여 주스, 젤리 등 다양한 제품을 개발할 필요도 있다. A hardy kiwifruit is a good source of phenolics and antioxidants. This study was aimed at investigating the effects of hot air drying at three different temperatures (35, 50, and 70℃) for 24 h on the total phenolic and flavonoid contents and the antioxidant capacity of hardy kiwifruits of Actinidia arguta × A. deliciosa cv. Mansoo. Dried kiwifruit extracts were produced using 80% (v/v) aqueous ethanol under homogenization. Hot air drying lowered the total phenolic and flavonoid contents and decreased the antioxidant capacity in the dried kiwifruits compared with their fresh counterparts. The results also revealed high positive linear correlations of antioxidant capacity with total phenolic and flavonoid contents, indicating that phenolics are the major contributors to antioxidant capacity. Our results suggested that an optimal drying process must be determined for industrial application in order to obtain dried kiwifruits with increased antioxidant capacity for consumers.

톳(Hizikia fusiformis)의 최종당화산물 저해 효능 및 신장 보호 효과

김민경 ( Mingyeong Kim ),조치흥 ( Chi Heung Cho ),염가희 ( Ga Hee Youm ),박용곤 ( Yongkon Park ),이상훈 ( Sang-hoon Lee ) 한국키틴키토산학회 2020 한국키틴키토산학회지 Vol.25 No.4

국내 자생 갈조류 중 하나인 톳을 이용하여 추출방법에 따른 생리활성 물질의 함량을 분석하였다. 또한 당뇨병성 신장증의 합병증을 유발하는 원인으로 알려진 최종당화산물과 콜라겐 사이의 교차결합 절단 및 교차결합 형성 억제 효과 및 최종당화산물의 생성 억제 효과를 평가하였다. 총 페놀, 탄닌 및 총 플라보노이드와 같은 생리활성 성분을 가장 많이 함유하는 추출 조건이 70% 에탄올임을 확인하였다. 톳 70% 에탄올 추출물(70% EtOH SFE)은 최종당화산물과 콜라겐 사이의 교차결합 절단 및 교차결합 형성 억제 및 최종당화산물의 생성을 억제하는 효능이 가장 우수하였다. 특히 70% EtOH SFE는 MGO에 의해 유도되는 mesangial 세포 내 산화적 스트레스를 유의적으로 감소시킬 뿐만 아니라 apoptosis로 인한 세포 사멸을 감소시켰다. 결과들을 종합해보면 톳은 부작용이 적은 당뇨합병증 예방 및 치료제 개발 후보물질로서 가능성이 충분한 것으로 판단된다. 향후 톳의 최종당화산물에 의한 당뇨합병증에 활성을 가지는 성분의 동정 및 당뇨 합병증 예방과 치료에 작용하는 메커니즘 규명 연구가 추가적으로 진행되어야 할 것으로 사료된다. In this study, we evaluated the anti-glycation effects and renal protective properties of Sargassum fusiforme extract against advanced glycation end-products (AGEs)-induced oxidative stress. Among the extracts of S. fusiforme with different concentrations of ethanol (0, 10, 30, 70, and 95%(v/v)), 70% aqueous ethanol extract of S. fusiforme (70% EtOH SFE) showed the highest total phenolics, condensed tannin, and total flavonoid contents (0.797±0.036 mg GAE/g DW, 0.298±0.009 mg CE/g DW, 0.381±0.060 mg CE/g DW, respectively). All tested extracts of S. fusiforme showed concentration-dependent anti-glycation activity, in particular, 70% EtOH SFE exhibited the most prominent anti-glycation effects in all in vitro experiments. 70% EtOH SFE inhibited 7.13% AGEs-BSA and collagen cross-link at a concentration of 100 μg/mL , which was significantly (p < 0.05) higher than aminoguanidine (36.9%.), the well-known cross-link inhibitor. 70% EtOH SFE (100 μg/mL) showed AGEs-BSA and collagen cross-link breaking activity to the 17.9%. Additionally, 70% EtOH SFE inhibited formation of AGEs in a concentration-dependent manner up to 66.8%. Furthermore, treatment with 70% EtOH SFE not only increased the viability of methylglyoxal (MGO)-induced mesangial cells to 64.9%, but also dramatically reduced the intracellular oxidative stress level to 94.9%. The treatment of 70% EtOH SFE was decreased MGO-induced apoptotic cell death by 21.9% while aminoguanidine by 33.4%. Consequently, our results suggest that 70% EtOH SFE may be a good source of natural therapeutic agent for management of diabetic complications via anti-glycation effect and renal protective ability against MGO-induced oxidative stress.