• 한국응용과학기술학회지
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• 제37권5호
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• pp.1088-1099
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• 2020

본 연구는 화장품 소재로서 양하의 가능성을 확인하기 위한 것이다. 이를 위해 우리는 양하추출물을 사용하여 항산화, 항염증, 주름개선 효과에 대한 생리 활성 평가를 실시하였다. 이 실험을 하기 위해, 양하꽃 추출물 (ZMF)과 양하잎 추출물 (ZML)을 70% 에탄올로 추출하였다. 항염증 효과를 알아보기 위해 macrophage (Raw 264.7)를 이용해 시료의 세포독성 평가와 nitric oxide 저해능을 측정하였다. ZMF과 ZML의 DPPH 라디칼 소거능, ABTS+ 라디칼 소거능, SOD) 유사 활성 측정 결과 농도 의존적으로 활성이 증가하였다. ZMF는 ABTS+ radical scavenging activity와 superoxide dismutase (SOD)-like activity 측정 결과 ZML보다 높은 항산화 활성을 보였다. NO 저해능 측정 결과에 따르면 ZMF는 농도 의존적으로 NO가 저해되어 우수한 항염증 효과를 나타냈다. ZMF의 pro-collagen type-1 합성량은 25 ㎍/ml에서 110% 이상의 우수한 효과를 나타내었으며, MMP-1 저해능은 25 ㎍/ml에서 20%의 활성을 나타냈다. 이 결과로 ZMF는 주름개선용 화장품 소재로 응용이 가능할 것으로 판단된다. 항산화, 항염증, 주름개선 평가 결과, 양하의 생리 활성 효과가 검증되었으므로 천연 화장품 재료로 사용될 수 있을 것으로 사료된다.

• 한국식품저장유통학회지
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• 제24권5호
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• pp.688-696
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• 2017

본 연구는 불등풀가사리로부터 분리된 조다당류와 산가수분해된 다당류의 분자량에 따른 이화학적 품질특성과 항산화 활성 및 피부 주름 개선 효과를 확인하였다. 산가수분해 분자량별 다당류의 수율은 100 kDa 이상에서 29.6%로 가장 높았으며, 10 kDa 이하에서 10.4%로 가장 낮았다. 이화학적 특성으로 총당, uronic acid, sulfate 함량은 100 kDa 이상에서 각각 85.82%, 32.85 g/100g 및 39.04%로 조다당에 비해 증가하였다. 색도은 분자량이 증가함에 따라 L 값, a 값 및 b 값이 감소하였으며, 점도는 분자량이 높을수록 증가하였다. ORAC 활성은 100 kDa 이상에서 $180.07{\mu}M$을 나타내어 조다당 $34.70{\mu}M$에 비해 항산화활성이 우수하였다. L132 세포 사멸에 대한 보호효과는 $H_2O_2$ 처리 구간대비 분자량 100 kDa 이상($0.5-5{\mu}g/mL$)에서 87.34-103.85%로 세포 활성이 증가하여 세포 보호효과를 나타내었다. MMP-1 활성은 세포에 UVB를 조사한 후 분자량 100 kDa 이상 $0.05-0.5{\mu}g/mL$ 농도에서 69.67-100.87%의 저해 활성을 나타내었다. 또한, pro-collagen 생합성 역시 UVB를 조사한 후 분자량 100 kDa 이상 $0.05-0.5{\mu}g/mL$ 농도에서 64.91-77.80%의 합성능을 나타내어 농도 의존적으로 증가하는 것을 확인하였다. 따라서, 불등풀가사리 유래 산가수분해된 다당류의 분자량에 따른 주름 개선 효과를 통해 피부 항노화 기능성 소재로 다양하게 활용 가능할 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제17권4호
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• pp.307-314
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• 2013

Connective tissue growth factor (CTGF) is a potent pro-fibrotic factor, which is implicated in fibrosis through extracellular matrix (ECM) induction in diabetic cardiovascular complications. It is an important downstream mediator in the fibrotic action of transforming growth factor ${\beta}$ ($TGF{\beta}$) and is potentially induced by hyperglycemia in human vascular smooth muscle cells (VSMCs). Therefore, the goal of this study is to identify the signaling pathways of CTGF effects on ECM accumulation and cell proliferation in VSMCs under hyperglycemia. We found that high glucose stimulated the levels of CTGF mRNA and protein and followed by VSMC proliferation and ECM components accumulation such as collagen type 1, collagen type 3 and fibronectin. By depleting endogenous CTGF we showed that CTGF is indispensable for the cell proliferation and ECM components accumulation in high glucose-stimulated VSMCs. In addition, pretreatment with the MEK1/2 specific inhibitors, PD98059 or U0126 potently inhibited the CTGF production and ECM components accumulation in high glucose-stimulated VSMCs. Furthermore, knockdown with ERK1/2 MAPK siRNA resulted in significantly down regulated of CTGF production, ECM components accumulation and cell proliferation in high glucose-stimulated VSMCs. Finally, ERK1/2 signaling regulated Egr-1 protein expression and treatment with recombinant CTGF reversed the Egr-1 expression in high glucose-induced VSMCs. It is conceivable that ERK1/2 MAPK signaling pathway plays an important role in regulating CTGF expression and suggests that blockade of CTGF through ERK1/2 MAPK signaling may be beneficial for therapeutic target of diabetic cardiovascular complication such as atherosclerosis.

• Biomolecules & Therapeutics
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• 제20권1호
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• pp.104-112
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• 2012

The fruit of Terminalia chebula Retzius has been used as a panacea in India and Southeast Asia but its biological activities have not been fully elucidated. Here we report anti-arthritic and analgesic effect of NDI10218, a standardized ethanol extract of Terminalia chebula, on collagen-induced arthritis and acetic acid-induced writhing model, respectively. Arthritis was induced in DBA/1J mice by immunizing bovine type II collagen and mice were treated with NDI10218 daily for 5 weeks after the onset of the disease. NDI10218 reduced the arthritis index and blocked the synovial hyperplasia in a dose-dependent manner. The serum levels of pro-inflammatory cytokines TNF-${\alpha}$, IL-6, and IL-$1{\beta}$ were significantly reduced in mice treated with NDI10218. Production of the inflammatory IL-17, but not immunosuppressive IL-10, was also inhibited in splenocytes isolated from NDI10218-treated arthritis mice. Administration of NDI10218 markedly decreased the number of T cell subpopulations in the regional lymph nodes of the arthritis mice. Finally, NDI10218 reduced the number of abdominal contractions in acetic acid-induced writhing model, suggesting an analgesic effect of this extract. Taken together, these results suggest that NDI10218 can be a new therapeutic candidate for the treatment of rheumatoid arthritis.

• 인간식물환경학회지
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• 제23권2호
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• pp.191-199
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• 2020

Noni has been used for medicinal purposes for more than 2,000 years in South Pacific Polynesia, China and India, and has been heavily ingested as an extract for its excellent antioxidant and anti-inflammatory effects. However, a recent study found that the noni extract causes digestive disorders, kidney problems, and liver diseases, which made it necessary to use it for other purposes than as an extract. In this study, we want to evaluate the potential of noni as an anti-oxidant, anti-inflammatory and anti-wrinkling agent. Methods: Noni was freeze-dried, extracted in water, and concentrated. Skin cells were treated with the noni extract for 24 hrs and then were exposed to UVB (55 mJ/cm²). After 48 hrs of incubation, pro-inflammatory cytokine, elastase, MMP-1 and type-1 procollagen levels were measured by ELISA. Results: To find out the antioxidant effect of the noni extract, the DPPH and ABTS radical scavenging activity experiments were conducted and the noni extract showed 97.0 % and 92.0 % antioxidant efficacy at 200 ㎍/mL respectively. The noni extract (50 and 100 ㎍/mL) decreased IL-6 and TNF-α in RAW 264.7 cells induced by LPS in a concentration-dependent manner. In the RT-PCR experiment involving NO production, the noni extract (50 and 100 ㎍/mL) inhibited NO production by strongly inhibiting iNOS mRNA expression, and also inhibited the elevation of MMP-1 and elastases caused by UVB irradiation by 25.0 % and 7.0 % respectively. In addition, type-1 procollagen was elevated by 20.0 % by the noni extract treatment in HaCaT cells. Conclusion: The noni extract has photoprotective ability by reducing proinflammatory mediators, elastase and MMP-1 production, and elevation of collagen synthesis. Our findings suggest that the noni extract might be a good natural substance to protect against UVB-induced premature skin aging.

• 대한한의학회지
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• 제43권3호
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• pp.1-15
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• 2022

Objectives: Myrrh have been used as a traditional remedy to treat infectious and inflammatory diseases. However, it is largely unknown whether myrrh ethanol extract could exhibit the inhibitory activities against particulate matter (PM)-induced skin injury on human keratinocytes, HaCaT cells. Therefore, this study was aimed to investigate the inhibitory activity of myrrh ethanol extract on PM-induced skin injury in HaCaT cells. Methods: To investigate the inhibitory effects of myrrh ethanol extract in HaCaT cells, the skin injury model of HaCaT cells was established under PM treatment. HaCaT keratinocyte cells were pre-treated with myrrh ethanol extract for 1 h, and then stimulated with PM. Then, the cells were harvested to measure the cell viability, reactive oxygen species (ROS), pro-inflammatory cytokines including interleukin (IL) 1-beta, IL-6, and tumor necrosis factor (TNF)-𝛼, hyaluronidase, collagen, MMPs. In addition, we examined the mitogen activated protein kinases (MAPKs) and inhibitory kappa B alpha (I𝜅-B𝛼) as inhibitory mechanisms of myrrh ethanol extract. Results: The treatment of myrrh ethanol extract inhibited the PM-induced cell death and ROS production in HaCaT cells. In addition, myrrh ethanol extract treatment inhibited the PM-induced elevation of IL-1beta, IL-6, and TNF-𝛼. Also, myrrh ethanol extract treatment inhibited the increase of hyaluronidase, MMP and decrease of collagen. Furthermore, myrrh ethanol extract treatment inhibited the activation of MAPKs and the degradation of I𝜅-B𝛼. Conclusions: Our result suggest that treatment of myrrh ethanol extract could inhibit the PM-induced skin injury via deactivation of MAPKs and nuclear factor (NF)-𝜅B in HaCaT cells. This study could suggest that myrrh ethanol extract could be a beneficial agent to prevent skin damage or inflammation.

• Restorative Dentistry and Endodontics
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• 제46권1호
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• pp.4.1-4.16
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• 2021

Objectives: This study evaluated the biocompatibility and bioactive potential of NeoMTA Plus mixed as a root canal sealer in comparison with MTA Fillapex. Materials and Methods: Polyethylene tubes filled with NeoMTA Plus (n = 20), MTA Fillapex (n = 20), or nothing (control group, CG; n = 20) were inserted into the connective tissue in the dorsal subcutaneous layer of rats. After 7, 15, 30 and 60 days, the specimens were processed for paraffin embedding. The capsule thickness, collagen content, and number of inflammatory cells (ICs) and interleukin-6 (IL-6) immunolabeled cells were measured. von Kossa-positive structures were evaluated and unstained sections were analyzed under polarized light. Two-way analysis of variance was performed, followed by the post hoc Tukey test (p ≤ 0.05). Results: At 7 days, the capsules around NeoMTA Plus and MTA Fillapex had more ICs and IL-6-immunostained cells than the CG. However, at 60 days, there was no significant difference in the IC number between NeoMTA Plus and the CG (p = 0.1137) or the MTA Fillapex group (p = 0.4062), although a greater number of IL-6-immunostained cells was observed in the MTA Fillapex group (p = 0.0353). From 7 to 60 days, the capsule thickness of the NeoMTA Plus and MTA Fillapex specimens significantly decreased, concomitantly with an increase in the collagen content. The capsules around root canal sealers showed positivity to the von Kossa stain and birefringent structures. Conclusions: The NeoMTA Plus root canal sealer is biocompatible and exhibits bioactive potential.

• 생명과학회지
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• 제26권1호
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• pp.34-41
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• 2016

학명이 Aronia melanocarpa인 블랙초크베리는 항산화, 항염증, 항암 효능이 뛰어난 것으로 보고되고 있다. 본 연구에서는 블랙초크베리에 대한 collagenase inhibition effects와 산화적 스트레스에 유도된 matrix metalloproteinase(MMP), MAPkinase 그리고 AP-1의 발현 그리고/또는 인산화와 같은 분자생물학적 메카니즘을 조사하였다. Collagenase inhibition 효과는 블랙초크베리 에틸아세테이트 분획물(AE)이 500 μg/ml의 농도에서 77.2% 이상의 저해효능을 나타내었고 이는 대조군인 Epigallocatechin gallate의 결과(500 μg/ml에서 83.9%)와 비교해서 유의할 만한 결과였다. Reactive oxygen species (ROS) assay는 AE에서 가장 농도의존적으로 ROS 생성이 감소되었고, 75 μg/ml의 농도에서 약 70%로 가장 낮은 활성산소가 생성되었다. MTT assay 결과, H₂O₂에 유도된 CCRF 세포에 AE를 처치하였을 때 농도의존적으로 세포 생존율이 증가하였다. 그리고 특히, AE는 H₂O₂에 유도된 CCRF 세포에서의 MMPs (MMP-1, -3 그리고 -9), MAPK (ERK, JNK 그리고 p38) 그리고 AP-1 (c-Fos와 c-Jun)의 발현과 인산화를 억제하였고, pro-collagen type I의 발현은 증가시켰다. 따라서 블랙초크베리 에틸아세테이트 분획물은 주름억제 및 콜라겐 생성의 효능이 있으며 기능성 식품 및 화장품 소재 개발 산업에서의 응용이 가능할 것으로 기대된다.

• 한국식품영양과학회지
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• 제46권5호
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• pp.562-571
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• 2017

본 연구에서는 선별된 유색미 5품종에 대한 자외선 조사에 따른 노화 억제 기전을 확인하고자 항산화 활성과 활성산소종(ROS) assay를 분석하였으며, 주름형성을 유발하는 효소인 MMPs와 그 upstream인 MAPK pathway(ERK, JNK 및 p38)에서의 인산화 발현을 얼마나 저해하는지를 분석하였다. 유색미 5품종 가운데 전자공여능과 ABTS 소거활성 결과, 신토흑미 에탄올 추출물(SRE)과 흑설 에탄올 추출물(HE), SRE와 흑설 열수 추출물(HW)이 각각 뛰어난 소거활성을 보였으며, collagenase 저해 활성 결과 HE가 전 농도에서 고르게 높은 저해효과를 나타내었다. 또한, ROS assay에서 SE와 HE가 전 농도에서 매우 효과적인 ROS 생성억제를 나타냄에 따라 우리는 선별된 유색미 가운데 HE를 이용하여 MMPs와 MAPK의 발현억제 pathway를 확인하였다. 그 결과 pro-collagen type I은 $100{\mu}g/mL$의 농도에서 발현이 증가하였고, MMP-1과 -13은 농도 의존적으로 발현이 억제되었으며, ERK와 p38의 인산화 또한 농도 의존적으로 억제하고 있음을 확인하였다. 이러한 결과에 따라 MMPs의 mRNA 발현을 확인하기 위한 RT-PCR을 실시하였으며, MMP-1과 3의 mRNA의 발현은 농도 의존적으로 감소하였고 MMP-9는 HE를 $100{\mu}g/mL$의 농도로 처치한 경우에 발현이 감소하였음을 확인하였다. 선별된 유색미 5종의 에탄올 추출물은 항산화 활성이 우수하였고, 특히 HE의 주름억제 활성 효능을 확인함에 따라 기능성 식품 화장품 분야에서 주름개선 및 항노화 소재로서의 가능성을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제25권7호
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• pp.1070-1083
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• 2015

A gene (GT-SM3B) encoding a thermostable secreted oligoendopeptidase (GT-SM3B) was cloned from the thermophile Geobacillus thermoleovorans DSM 15325. GT-SM3B is 1,857 bp in length and encodes a single-domain protein of 618 amino acids with a 23-residue signal peptide having a calculated mass of 67.7 kDa after signal cleavage. The deduced amino acid sequence of GT-SM3B contains a conservative zinc metallopeptidase motif (His⁴⁰⁰-Glu⁴⁰¹-X-XHis⁴⁰⁴). The described oligopeptidase belongs to the M3B subfamily of metallopeptidases and displays the highest amino acid sequence identity (40.3%) to the oligopeptidase PepF_Ba from mesophilic Bacillus amyloliquefaciens 23-7A among the characterized oligopeptidases. Secretory production of GT-SM3B was used, exploiting successful oligopeptidase signal peptide recognition by Escherichia coli BL21 (DE3). The recombinant enzyme was purified from the culture fluid. Homodimerization of GT-SM3B was determined by SDS-PAGE. Both the homodimer and monomer were catalytically active within a pH range of 5.0–8.0, at pH 7.3 and 40℃, showing the K_m, V_max, and k_cat values for carbobenzoxy-Gly-Pro-Gly-Gly-Pro-Ala-OH peptidolysis to be 2.17 ± 0.04 × 10^-6 M, 2.65 ± 0.03 × 10^-3 µM/min, and 5.99 ± 0.07 s^-1, respectively. Peptidase remained stable at a broad pH range of 5.0–8.0. GT-SM3B was thermoactive, demonstrating 84% and 64% of maximum activity at 50℃ and 60℃, respectively. The recombinant oligopeptidase is one of the most thermostable M3B peptidase, retaining 71% residual activity after incubation at 60℃ for 1 h. GT-SM3B was shown to hydrolyze a collagenous peptide mixture derived from various types of collagen, but less preferentially than synthetic hexapeptide. This study is the first report on an extracellular thermostable metallo-oligopeptidase.