감 심지 에탄올 추출물의 항산화 활성 및 신경세포 보호 효과

변의백,김민진,김순정,오남순,박상현,김우식,송하연,한정무,김광욱,변의홍 한국식품과학회 2020 한국식품과학회지 Vol.52 No.1

This study examined the antioxidant activity and neuroprotective effects of ethanol extracts obtained from Diospyros kaki core (DCE). The total polyphenol and flavonoid contents in DCE was 786.47±15.27 and 31.14±0.82 mg/ g, respectively. In addition, DCE exhibited a dose-dependent induction of radical scavenging activity, determined by 1,1- diphenyl-picrylhydrazyl (DPPH), 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonicacid) (ABTS), ferric reducing antioxidant power (FRAP), and reducing power assays. The viability of HT22 hippocampal cells was examined to investigate the neuroprotective effect of DCE. DCE treatment did not induce cytotoxicity at concentrations below 1,000 μg/mL. Additionally, DCE treatment in the background of H2O2 induce oxidative stress revealed a significant increase in the survival rat, indicated by increased SOD activity and decreased levels of MDA, a lipid peroxidation product. Therefore, the results suggest that DCE can be used as a source of natural antioxidants source and a therapeutic agent for the treatment of brain disorders induced by oxidative stress and neuronal damage.

꾸지뽕(Cudrania tricuspidata) 열매에서 추출된 조다당의 대식세포 보호 효과

변의백,김광욱,오남순,박원종,박상현,김우식,송하연,한정무,변의홍 한국식품영양과학회 2019 한국식품영양과학회지 Vol.48 No.12

It is believed that Cudrania tricuspidata fruits polysaccharides extracts (CTPS) have antioxidant, anti-inflammatory, anti-cancer, and immune activity effects. However, there have been no studies that have shown the cytoprotective effects of CTPS. Therefore, the present study was conducted to confirm the cytoprotective effect of CTPS in terms of their inhibition of cytotoxicity, caspase activity, reactive oxygen species (ROS) production, and mitochondrial outer membrane permeability (MOMP) increase in Raw264.7. Macrophage apoptosis was induced by staurosporine (STS) treatment. CTPS treatment significantly reduced STS induced cell death and annexin V positive cell levels in a dose-dependent manner. In addition, CTPS treatment inhibited activation of caspase-3, 8, 9, and PARP in macrophages. Further, CTPS treatment reduced STS induced ROS production and MOMP increase in macrophages. In conclusion, CTPS treatment inhibits STS-induced macrophage apoptosis through inhibition of cytotoxicity, caspase activation, ROS production, and MOMP increase. These results suggest CTPS could be a potential candidate for treatment of immune cell protection effects. 본 연구에서는 꾸지뽕 열매 조다당 추출물(CTPS)이 세포 생존율에 치중했던 이전 세포 보호 효과 연구와는 다르게 대식세포의 세포 보호 효과를 유도하는지 알아보기 위하여 caspase 세포사멸 신호전달과 산화적 스트레스에 관하여 연구하였다. 먼저, 대식세포에 CTPS를 처리한 후 세포독성물질(staurosporine)을 병용 처리하여 세포 보호 효과를 유도하는지 평가해보았다. CTPS가 세포독성물질로 인해 일어나는 세포사멸을 억제시키는 것으로 관찰되었고, 세포독성물질로 인한 caspase 의존적인 세포사멸도 CTPS에 의해 억제되는 것을 관찰할 수 있었다. 게다가 세포독성물질로 인해 생성된 활성산소종이 CTPS에 의해 억제되는 것을 관찰하였으며, 세포독성물질로 인해 증가한 미토콘드리아 외막 투과성도 CTPS에 의해 감소하는 것을 관찰할 수 있었다. 결론적으로 CTPS가 세포독성물질로 인한 caspase 활성을 억제하고 산화적 스트레스를 감소시킴으로써 대식세포의 보호 효과를 나타내며, 이는 CTPS가 앞으로 세포 보호 효과 소재로 개발되어 인체 내 면역세포 보호 효과에 많은 기여를 할 것으로 판단된다.

β-(1,3)-Glucan Isolated from Agrobacterium Species Induces Maturation of Bone Marrow-derived Dendritic Cells and Drives Th1 Immune Responses

변의백,박상현,성낙윤,변의홍,박철환 한국식품과학회 2015 Food Science and Biotechnology Vol.24 No.4

The β-(1,3)-glucan originally isolated from Agrobacterium species was investigated for prompting maturation of dendritic cells (DCs) and driving Th1 immune responses. Bone-marrow derived DCs separated from mice were analyzed for augmentation of cell surface molecule (CD80, CD86, and major histo-compatability complex (MHC) class I/II) expression and pro-inflammatory cytokine production (tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6). β-Glucan functionally induced DCs activation via augmentation of CD80, CD86, and MHC class I/II expression and cytokine production (TNF-α, IL-1β, and IL-6). β-Glucan induced secretion of IL-12p70, but not IL-10. Both mitogen-activated protein kinases and nuclear factor (NF)-κB signaling mediated production of pro-inflammatory cytokine induced by β- glucan. β-Glucan-treated DCs exhibited accelerated proliferation of murine splenocytes with increased levels of interferon (IFN)-γ. β-Glucan regulates innate and adaptive immunity via DCs activation and Th1 polarization of immune responses.

Gamma-Irradiated Luteolin Inhibits 3-Isobutyl-1-Methylxanthine-Induced Melanogenesis Through the Regulation of CREB/MITF, PI3K/Akt, and ERK Pathways in B16BL6 Melanoma Cells

변의백,송하연,Sajid Mushtaq,김혜민,강정애,양미소,성낙연,장범수,변의홍 한국식품영양과학회 2017 Journal of medicinal food Vol.20 No.8

Luteolin was gamma irradiated at doses of 0, 15, 30, 50, 70, and 100 kGy. We observed that the luteolin peak decreased simultaneously with the appearance of new radiolytic peaks, using high-performance liquid chromatography (HPLC). The highest new radiolytic peak gamma-irradiated luteolin (GLM) was observed at a dose of 70 kGy, and the GLM was identified by nuclear magnetic resonance and high-performance-liquid-chromatography-quadrupole-time-of-flight (HPLC-Q-TOF) mass spectrometry. We examined whether 70 kGy gamma-irradiated luteolin has more effective anti-melanogenic effects than intact luteolin. Seventy kilograys of gamma-irradiated luteolin inhibited melanin synthesis and intracellular tyrosinase activity without cytotoxicity, whereas the intact luteolin-treated group did not show anti-melanogenic activity in 3-isobutyl-1-methylxanthine-stimulated B16BL6 melanoma cells. The expression of melanogenic enzymes, such as tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2, was decreased by 70 kGy gamma-irradiated luteolin treatment, owing to the suppression of microphthalamia-associated transcription factor and 3′,5′-cyclic adenosine monophosphate (cAMP) response element binding protein. In addition, gamma-irradiated luteolin decreased the phosphorylation of phosphoinositide 3-kinase (PI3K)/Akt and extracellular regulated kinase (ERK). The anti-melanogenic effects of 70 kGy gamma-irradiated luteolin were attenuated by the treatment of two specific inhibitors (PD98059 and LY294002), and these results indicate that the anti-melanogenic effects were mediated by ERK and PI3K signaling pathways. Therefore, our findings suggest that gamma-irradiated luteolin can be a potential cosmeceutical agent for skin whitening.

감 심지 에탄올 추출물의 미백 활성

변의백,민희숙,김민진,김순정,박원종,송하연,한정무,김광욱,변의홍 한국식품영양과학회 2019 한국식품영양과학회지 Vol.48 No.7

This study examined the whitening effect of a Diospyros kaki core ethanol extract (DCE). DCE was used to evaluate its inhibitory effects on tyrosinase and melanin synthesis. The mushroom tyrosinase inhibitory activity of DCE increased in a dose dependent fashion. DCE treatment did not exert cytotoxicity at doses ranging from 62.5 to 1,000 μg/mL in B16F10 melanoma cells. Treatment of 3-isobutyl-1-methylxanthine strongly induced melanin synthesis (518.18±9.1%) as compared with the control group (100%), while the DCE-treated group showed effective inhibition of melanin synthesis (500 and 1,000 μg/mL, 222.73±31.8% and 177.27±4.5%, respectively). The anti-melanogenic effects of DCE were mediated by the regulation of melanogenesis-related proteins, including cAMP response element-binding protein, microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2. Therefore, these results suggest that DCE has a whitening effect, and it could be developed as a whitening ingredient in cosmetics. 본 연구에서는 감(Diospyros kaki) 심지의 미백 효과를 조사하고자 에탄올을 이용하여 추출하고 감 심지 에탄올 추출물(DCE)의 mushroom tyrosinase 저해 활성을 측정한 결과 농도 의존적으로 저해 활성이 증가한 것을 확인하였다. 세포 내 미백 효과를 알아보기 위하여 B16F10 melanoma 세포를 사용하여 세포 생존율 및 세포 내 멜라닌 생합성 억제 효과를 측정한 결과, 세포독성 없이 세포 내 멜라닌 합성을 유의적으로 억제함을 확인하였다. 마지막으로 세포 내 미백 메커니즘을 확인한 결과 DCE 처리가 p-CREB, MITF, tyrosinase, TRP-1 및 TRP-2의 단백질 발현을 감소시키는 것을 확인하였다. 이러한 결과를 바탕으로 DCE는 천연 미백 기능성 소재로 활용될 수 있을 것으로 사료된다.

방사선분자변환기술 기반 천연 생물소재 구조변환에 따른가공적성 및 생리활성 증진 연구

변의백,송하연,김혜민,김우식,이승식,최대성,임상용,정병엽 한국식품과학회 2018 한국식품과학회지 Vol.50 No.3

Radiation technology (RT) has long been applied in various fields for increasing the safety and shelf-life of foods by controlling pathogen-induced poisoning. RT was introduced for the first time in Korea in the 1950s to eliminate harmful microorganisms in food materials. In the 1980s, RT had been scientifically proven to be effective for the sterilization of food and public health products. In recent years, irradiation with gamma rays has also been used for improving physiological properties through the structural modification of natural molecules, which has been proposed to be applicable to various industries. In particular, radiation transformation technology (RTT), which involves the development of new functional compounds through the molecular conversion of natural biomaterials, is becoming a new high-value technology as a fusion technique of RT and biotechnology. The present reports have suggested that RTT can be an effective tool for the development of new functional compounds and improvement of the physiological activity of biomolecules.

특수목적식품 제조용 프레스햄의 위생화를 위한 제조공정 단계별 감마선 조사기술 적용 : 미생물학적 평가

변의백,이주운,김재훈,오상희,서지현,김장호,변명우 한국축산식품학회 2005 심포지움 및 학술발표회 Vol.- No.35

본 연구는 특수목적식품을 개발하기 위한 대표적인 비가열 살균처리 기술인 방사선 살균기술을 이용하여 저장 중 프레스햄의 미생물학적 품질을 평가하고자 하였다. 그 결과 감마선비조사구의 경우 제조공정 단계에 관계없이 30℃ 저장 4일 째에 모두 미생물 생육이 활발해져 장기저장에 부적합한 것으로 나타났으나, 감마선을 조사할 경우 훈연후 및 살균전 시료의 경우 5 kGy, 살균후 시료의 경우 3 kGy의 감마선 조사로 30℃ 저장 4일 까지 미생물의 생육이 검출되지 않았다. 따라서, 감마선 조사기술은 프레스햄의 살균방법으로 이용이 가능하며, 특히 장기저장을 목적으로 하는 우주식, 환자식, 전투식량 등의 특수목적식품의 개발에 그 이용이 가능하다 할 수 있다.

Procyanidin C1 Causes Vasorelaxation Through Activation of the Endothelial NO/cGMP Pathway in Thoracic Aortic Rings

변의백,성낙윤,양미소,송두섭,변의홍,김재경,박종흠,송범석,이주원,박상현,변명우,김재훈 한국식품영양과학회 2014 Journal of medicinal food Vol.17 No.7

The aim of this study was to clarify the efficacy of procyanidin C1 (Pro C1) for modulating vascular tone. Pro C1 induced a potent vasorelaxant effect on phenylephrine-constricted endothelium-intact thoracic aortic rings, but had no effect on denuded thoracic aortic rings. Moreover, Pro C1 caused a significant increase in nitric oxide (NO) production in endothelial cells. Pro C1-induced vasorelaxation and Pro C1-induced NO production were significantly decreased in the presence of a nonspecific potassium channel blocker (tetraethylammonium chloride [TEA]), an endothelial NO synthase inhibitor (NG-monomethyl-L-arginine [L-NMMA]), and a store-operated calcium entry inhibitor (2-aminoethyl diphenylborinate [2-APB]). Pro C1-induced vasorelaxation was also completely abolished by an inhibitor of soluble guanyl cyclase, which suggests that the Pro C1 effects observed involved cyclic guanosine monophosphate (cGMP) production. Interestingly, Pro C1 significantly enhanced basal cGMP levels. Taken together, these results indicate that Pro C1-induced vasorelaxation is associated with the activation of the calcium-dependent NO/cGMP pathway, involving potassium channel activation. Thus, Pro C1 may represent a novel and potentially therapeutically relevant compound for the treatment of cardiovascular diseases.

꾸지뽕(Cudrania tricuspidata) 잎으로부터 분리된 다당류 추출물의 면역 활성

변의백(Eui-Baek Byun),장범수(Beom-Su Jang),성낙윤(Nak-Yun Sung),변의홍(Eui-Hong Byun) 한국식품영양과학회 2016 한국식품영양과학회지 Vol.45 No.8

본 연구는 꾸지뽕 잎으로부터 추출된 다당류인 CTP의 처리가 면역세포의 활성에 미치는 영향에 관하여 평가하였다. CTP는 에탄올 침전법에 의하여 추출하였고, 면역 활성능의 평가는 대식세포주인 RAW 264.7 세포와 미분화 골수세포로부터 유도 분화시킨 대식세포 및 마우스 비장으로부터 유리시킨 비장세포에 CTP를 농도별로 처리하여 관찰하였다. 선천면역계에서 중요한 역할을 수행하는 대식세포에 CTP를 처리하였을 때 세포 증식률, NO 및 cytokine 분비능이 CTP 처리 농도 의존적으로 증가하는 것으로 관찰되었을 뿐만 아니라 비장세포에서도 이와 유사하게 세포 증식률이 증가하고 Th 1 type의 cytokine 분비능 또한 유의적으로 증가하는 것으로 관찰되었다. 이상의 결과로 미루어보아 꾸지뽕 다당류 추출물인 CTP는 다양한 면역세포의 활성을 증가시키는 것으로 생각하며 이를 활용하여 다양한 식품 및 건강보조식품을 개발한다면 그 경제적 가치가 매우 클 것으로 생각한다. The objective of this study was to evaluate the immunomodulatory activity of crude polysaccharide separated from Cudrania tricuspidata leaf. C. tricuspidata polysaccharide (CTP) was extracted by ethanol precipitation. Immunomodulation activity was tested in macrophage cells (RAW 264.7 and bone-marrow derived macrophage) and splenocytes. CTP treatment significantly increased cell proliferation up to 250 μg/mL in both RAW 264.7 and bone-marrow derived macrophages. In this concentration range (below 250 μg/mL), nitric oxide and cytokine [tumor necrosis factor (TNF)-α and interleukin (IL)-6] production also significantly increased. Similarly, splenocyte proliferation dosedependently increased except for the 1,000 μg/mL treated group. Regarding cytokine production activity in splenocytes, CTP treatment significantly increased production of Th 1 type cytokines [interferon (IFN)-γ] production but not Th2 type cytokines (IL-4). Therefore, the results indicate that CTP may have a potential effect on immunomodulatory activity in various immune cells, and this is useful for development of immune enhancing adjuvant materials as a natural ingredient.

그라비올라(Annona muricata) 잎에서 분리된 다당류 추출물의 큰포식세포 보호 효과

변의백(Eui-Baek Byun),김광욱(Kwangwook Kim),김우식(Woo Sik Kim),변의홍(Eui-Hong Byun) 한국식품영양과학회 2020 한국식품영양과학회지 Vol.49 No.2

본 연구에서는 항암, 항산화 및 항염증 활성을 일으키는 그라비올라 잎으로부터 추출한 다당류(ALPS)가 선천면역 및 적응면역과 밀접한 연관이 있는 큰포식세포의 보호 효과를 유도하는지 알아보기 위하여 큰포식세포에 ALPS를 처리한 후 세포독성물질(staurosporine)을 병용 처리하여 큰포식세포 보호 효과를 유도하는지 평가해보았다. 세포독성물질로 인해 일어나는 세포사멸이 ALPS에 의해 완화되는 것을 확인하였으며, 세포독성물질로 인한 caspase 의존적인 세포사멸이 ALPS에 의해 완화되는 것을 확인할 수 있었다. 또한 ALPS는 세포독성을 일으키는 활성산소종을 완화하는 것을 확인하였으며, 세포독성물질로 인해 증가한 미토콘드리아 외막 투과성이 ALPS에 의해 감소하는 것을 확인할 수 있었다. 따라서 이러한 결과들로 인해 외부의 독성물질로 인한 큰포식세포의 세포독성에 ALPS가 세포 보호 효과를 나타낸다고 판단되며, 이는 ALPS가 앞으로 세포 보호 효과소재로 개발될 가능성이 월등히 높다는 것을 보여주고 있다. This study focused on the macrophage protection effects of Annona muricata leaf polysaccharides (ALPS). Macrophage apoptosis was induced by a staurosporine (STS) treatment. STS induced macrophage apoptosis because of the cytotoxicity, caspase activation, reactive oxygen species (ROS) production, and increase in mitochondrial outer membrane permeability (MOMP). The inhibition of macrophage apoptosis by the ALPS treatment was confirmed. The ALPS treatment reduced STS-induced cell death and Annexin V positive cell levels significantly in a dose-dependent manner. In addition, the ALPS treatment inhibited the activation of caspase-3, 8, 9, and PARP in macrophages. Furthermore, the ALPS treatment reduced the STS-induced ROS production and MOMP increase in macrophages. Therefore, the ALPS treatment inhibits STS-induced macrophage apoptosis through the inhibition of cytotoxicity, caspase activation, ROS production, and MOMP increase, suggesting that ALPS could be a potential candidate as an immune cell protection drug.