• 한국식품과학회지
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• 제50권4호
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• pp.444-450
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• 2018

본 연구는 꿀벌 꽃가루 추출물(BPW)의 면역 활성에 관하여 알아보기 위하여, 선천면역계의 대표적인 수지상세포와 후천면역계의 대표적인 비장세포에 BPW를 처리 하여 면역세포의 활성능을 관찰하였다. 수지상 세포에 BPW를 처리하여 세포 생존율, 산화질소(II)와 사이토카인($TNF-{\alpha}$, IL-6과 $IL-1{\beta}$) 분비능과 세포 표면분자를 관찰 하였다. 세포 생존율은 수지상 세포에 BPW를 처리하였을 때, 세포 독성을 일으키지 않았으며 주요 면역 활성 인자인 산화질소(II) 분비능을 관찰한 결과, 농도 의존적으로 증가하는 것을 확인하였다. 또한 사이토카인의 분비능을 관찰한 결과, $TNF-{\alpha}$, IL-6과 $IL-1{\beta}$의 함량이 농도 의존적으로 증가하는 것으로 관찰되었다. 또한 활성화된 면역세포의 세포 표면에서 발현되는 CD80과 CD86의 발현과 항원제시에 밀접한 관련이 있는 MHC class I, II의 발현이 유의적으로 증가하는 것으로 관찰되었다. 또한 후천면역에서 중요한 역할을 수행하는 면역 T 세포가 다량 분포하는 지라 세포를 분리하여 BPW를 처리 하였을 때 Th1 세포가 분비하는 사이토카인의 함량이 농도 의존적으로 증가되는 것으로 나타났다. 이러한 결과로 미루어 볼 때 BPW는 선천면역뿐만 아니라 후천면역에 관여하는 다양한 면역세포의 활성화에 직간접적으로 관여하는 것으로 사료된다.

• 생명과학회지
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• 제19권2호
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• pp.163-168
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• 2009

천초근은 전통적으로 동양의학에서 항암제로 사용되어왔는데 인간 급성 백혈병 세포주인 Jurkat T 세포를 사용하여 천초근의 세포독성기작을 알아보았다. 천초근 뿌리(3 kg)를 메탄올로 추출, 증류한 후 물에 녹여 다시 dichloromethane으로 추출분획 하였다. 세포독성 활성을 보이는 dichloromethane 추출물을 연속적으로 HPLC를 통해 분리하였고 그 활성물질(65 mg)을 CCH1이라 명명하였다. CCH1을 0.5 ${\mu}g$/ml에서 2.0 ${\mu}g$/ml의 농도로 처리하고 세포사멸 과정을 보았다. 즉, mitochondria cytochrome c 방출, casapase-8, -9 및 caspase-3의 활성화, PARP 분해, DNA 단편화 현상들이 일어나는 것을 관찰하였다. 하지만, mitochondria cytochrome c 방출 억제자인 Bcl-xL이 과발현되는 Jurkat T 세포에서는 세포사멸현상이 일어나지 않았다. 이러한 결과는 CCH1이 mitochondria 의존적인 신호전달 과정을 통해서 세포사멸을 유도 한다고 할 수 있다. 그리고 CCH1에 의한 세포독성은 혈액에서 분리한 단핵구 세포보다 Jurkat T 세포에서 보다 강한 활성을 보였다.

• 생명과학회지
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• 제28권6호
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• pp.738-744
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• 2018

제주조릿대는 한라산 일대에서 자생하는 제주특산식물이다. 예로부터 조릿대 잎은 다양한 약리효과를 가지고 있어 전통의약에서 사용되어 왔을 뿐만 아니라 최근에는 식품 및 화장품 소재로서 활용되고 있다. 우리 연구팀은 제주조릿대 잎의 효율적인 산업적 활용을 촉진하기 위하여 열수 추출한 후 남는 잔사로부터 식물화합물을 다량으로 함유한 잔사 추출물(PRE)을 제조하는 방법을 보고한 바 있다. 본 연구는 PRE 및 분획물이 항산화 혹은 항염소재로서 활용 가능성을 평가하기 위하여 수행하였다. PRE 에틸아세테이트 분획물(EPRE)은 DPPH, ABTS, superoxide 자유기 소거활성이 우수하였고, LPS로 자극한 RAW 264.7 세포에서 활성 산소종(ROS)과 nitric oxide (NO) 생성을 효과적으로 억제하였다. EPRE는 nuclear factor E2-related factor 2 (Nrf 2)의 핵내 이동을 증가시킴으로써 heme oxygenase-1 (HO-1)의 발현을 유도하였다. HO-1 저해제인 zinc protoporphyrin (ZPP)은 EPRE에 의한 NO 생성 저해 능을 감소시키기 때문에 HO-1이 NO 생성 억제에 관여함을 알 수 있었다. 본 연구결과는 PRE와 EPRE가 항산화 및 항염 소재로서의 활용 가능성을 제시해 준다.

• 동의생리병리학회지
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• 제27권5호
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• pp.625-630
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• 2013

Erectile dysfunction (ED) is a highly prevalent disorder that affects millions of men worldwide. ED is now considered an early manifestation of atherosclerosis, and consequently, a precursor of systemic vascular disease. Lycii fructus extracts (LFE) were administered for 4 weeks to assess the improving effects on ED. Animals were divided into one normal group and four LFE-treated groups (0, 0.3, 0.6, and 1.2 g/kg). We induced ED in the study animals by oral administration of 20% ethanol instead of water everyday for 4 weeks. This study was designed to investigate the effects of LFE on the mRNA levels of inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS) expression; NO levels of nitric oxide (NO) and cyclic guanosine monophosphate (cGMP); blood profile; and erectile response of the corpus cavernosum of the rat penis. The libido of the LFE-administered male rats was higher than that of the ethanol control group. The erectile response of the corpus cavernosum was restored after LFE administration, to a level similar to the normal group. In addition, the iNOS in the corpus cavernosum of the male rats administered LFE decreased. In contrast, compared to the control group, LFE-administered male rats showed increased eNOS, NO and cGMP levels in the corpus cavernosum. These results indicate that LFE effectively restored ethanol-induced ED in male rats.

• 대한한의학방제학회지
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• 제22권1호
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• pp.151-165
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• 2014

Objectives : The aim of this study was to evaluate the efficacy of Aconiti Lateralis Preparata Radix (AP) and Acanthopanacis Cortex (AT) extracts in bone-derived adipocyte OP9 cell, osteoclast and osteoblast-like MG63 cells. Methods : MTT assay was used to evaluate the cytotoxicity of AP and AT extracts on OP9, osteoclast and MG63 cells. OP9 cells were treated with AP and AT, and the alterations in fat storage in the cells were determined by the Oil red O. To explain effects of RANKL-induced osteoclast differentiation in bone marrow macrophages, we performed the TRAP staining. The protein level of CAAAT/enhancer binding protein alpha ($C/EBP{\alpha}$) and peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$) as a adipocyte differentiation marker, and adiponectin was examined using western blot in differentiated OP9 cells. Effects of related genes were confirmed by luciferase assay using reporter assay. Results : AP and AT was not toxic on OP9 and MG63 cells, but AT was a little cytotoxic to osteoclast at the dose of $100{\mu}g/m{\ell}$. They could inhibit differentiation of OP9 cells and osteoclast with results of oil red O staining and TRAP staining. By western blot, AP and AT decreased the expression of $PPAR{\gamma}$ and $C/EBP{\alpha}$ which is the key transcription factor in adipogenesis and adiponectin secretion. AT also inhibited the BMP-4 activity in luciferase assay. AP also inhibited BMP-4 and Wnt3a activity, stimulated ER-${\beta}$ activity but inhibited androgen receptor activity. Conclusions : These results show AP and AT can be useful in osteoporosis and obesity via inhibition of osteoclast and adipocyte differentiation.

• 동의생리병리학회지
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• 제18권2호
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• pp.507-516
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• 2004

Alzheimer's disease(AD) is a geriatric dementia that is widespread in old age. In the near future AD will be the commom disease in public health service. Although a variety of oriental presciptions in study POD(Polygala tenuifolia extracted from dichlorometan) have been traditionally utilized for the treatment of AD, their pharmacological effects and action mechanisms have not yet fully elucidated. It has been widely believed that AP peptide divided from APP causes apoptotic neurotoxicity in AD brain. However, recent evidence suggests that CT105, carboxy terminal 105 aminoacids peptide fragment of APP, may be an important factor causing neurotoxicity in AD. SK-N-SH cells expressed with CT105 exhibited remarkable apoptotic cell damage. Based on morphological observations by phase contrast microscope and NO formation in the culture media, the CT105-induced cell death was significantly inhibited by POD. In addition, AD is one of brain degeneration disease. So We studied on herbal medicine that have a relation of brain degeneration. From old times, In Oriental Medicine, PO water extract has been used for disease in relation to brain degeneration. We were examined by ROS formation, neurite outgrowth assay and DPPH scravage assay. Additionally, we investigated the association between the CT105 and neurite degeneration caused by CT105-induced apoptotic response in neurone cells. We studied on the regeneratory and inhibitory effects of anti-Alzheimer disease in pCT105-induced neuroblastoma cell lines by POD. Findings from our experiments have shown that POD inhibits the synthesis or activities of CT105, which has neurotoxityies and apoptotic activities in cell line. In addition, treatment of POD(>50 ㎍/㎖ for 12 hours) partially prevented CT(105)-induced cytotoxicity in SK-N-SH cell lines, and were inhibited by the treatment with its. POD(>50 ㎍/㎖ for 12 hours) repaired CT105-induced neurite outgrowth when SK-N-SH cell lines was transfected with CT105. As the result of this study, In POD group, the apoptosis in the nervous system is inhibited, the repair against the degerneration of Neuroblastoma cells by CT105 expression is promoted. Decrease of memory induced by injection of scopolamin into rat was also attenuted by POD, based on passive avoidance test. Taken together, POD exhibited inhibition of CT105-induced apoptotic cell death. POD was found to reduce the activity of AchE and induced about the CA1 in rat hippocampus. Base on these findings, POD may be beneficial for the treatment of AD.

• 한국잠사곤충학회지
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• 제52권1호
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• pp.1-5
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• 2014

피부는 자외선과 같은 외부자극으로부터 피부를 보호하기 위해 멜라닌을 생성한다. 하지만 과도한 멜라닌의 생성은 기미, 주근깨와 같은 색소 침착을 유발하며 미관상 좋지 않을 뿐 아니라 피부 질환을 유발할 수 있다. 따라서 본 연구에서는 왕지네 추출물이 멜라닌 생성에 영향을 미치는지 여부를 B16/F10 melanoma 세포를 이용한 실험을 통해 확인하였다. 그 결과 유기용매를 이용한 왕지네 추출물의 멜라닌 생성량은 무처리 대비 36 ~ 67%로서 멜라닌 생성이 감소되는 것을 확인할 수 있었으며 특히 70% 에탄올을 이용한 왕지네 추출물의 멜라닌 생성량은 36%로 실험을 진행했던 추출물들 중에서 가장 낮음을 확인하였다. 이는 세포내 티로시나아제 효소 활성 측정에서도 유사한 결과를 얻었으며 이에 70% 에탄올로 추출한 왕지네 추출물은 피부 미백 기능성소재로서의 가능성이 높을 것으로 사료된다. 또한 티로시나아제 효소 활성을 단백질 수준에서 검정한 결과, 앞선 실험에서 티로시나아제 효소 활성이 가장 낮았던 70% 에탄올로 추출한 왕지네 추출물의 경우 세포내 glycosylation 된 티로시나아제의 발현이 감소함을 확인할 수 있었으며, 이는 70% 에탄올로 추출된 왕지네 추출물이 세포내 티로시나아제의 glycosylation을 억제하여 결과적으로 멜라닌 생성을 저해하는 것으로 추측할 수 있다.

• 한국작물학회지
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• 제37권3호
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• pp.288-298
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• 1992

Since the major important factors limiting plant growth and crop productivity are environmental stresses, of which low temperature is the most serious. It has been well known that many physiological processes are alterant in response to the environmental stress. With regard to the relationship between plant hormones and the regulation of chilling tolerance in rice seedlings, the major physiological roles of plant hormones: abscisic acid, ethylene and polyamines are evaluated and discussed in this paper. Rice seedlings were grown in culture solution to examine the effect of such plant hormones on physiological characters related to chilling tolerance and also to compare the different responses among tested cultivars. Intact seedlings about 14 day-old were chilled at conditions of 5$^{\circ}C$ and 80% relative humidity for various period. Cis-(＋)-ABA content was measured by the indirect ELISA technique. Polyamine content and ethylene production in leaves were determined by means of HPLC and GC respectively. Chilling damage of seedlings was evaluated by electrolyte leakage, TTC viability assay or servival test. Our experiment results described here demonstrated the physiological functions of ABA, ethylene, and polyamines related to the regulation of chilling tolerance in rice seedlings. Levels of cis-(＋)-ABA in leaves or xylem sap of rice seedlings increased rapidly in response to 5$^{\circ}C$ treatment. The tolerant cultivars had significant higher level of endogenous ABA than the sensitive ones. The ($\pm$)-ABA pretreatment for 48 h increased the chilling tolerance of the sensitive indica cultivar. One possible function of abscisic acid is the adjustment of plants to avoid chilling-induced water stress. Accumulation of proline and other compatible solutes is assumed to be another factor in the prevention of chilling injuies by abscisic acid. In addition, the expression of ABA-responsive gene is reported in some plants and may be involving in the acclimation to low temperature. Ethylene and its immediate precusor, 1-amincyclopropane-1-carboxylic acid(ACC) increased significantly after 5$^{\circ}C$ treatment. The activity of ACC synthase which converts S-adenosylmethionine (SAM) to ACC enhanced earlier than the increase of ethylene and ACC. Low temperature increased ACC synthase activity, whereas prolonged chilling treatment damaged the conversion of ACC to ethylene. It was shown that application of Ethphon was beneficial to recovering from chilling injury in rice seedlings. However, the physiological functions of chilling-induced ethylene are still unclear. Polyamines are thought to be a potential plant hormone and may be involving in the regulation of chilling response. Results indicated that chilling treatment induced a remarkable increase of polyamines, especially putrescine content in rice seedlings. The relative higher putrescine content was found in chilling-tolerant cultivar and the maximal level of enhanced putrescine in shoot of chilling cultivar(TNG. 67) was about 8 folds of controls at two days after chilling. The accumulation of polyamines may protect membrane structure or buffer ionic imbalance from chilling damage. Stress physiology is a rapidly expanding field. Plant growth regulators that improve tolerance to low temperature may affect stress protein production. The molecular or gene approaches will help us to elucidate the functions of plant hormones related to the regulation of chilling tolerance in plants in the near future.

• 한국식품영양학회:학술대회논문집
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• 한국식품영양학회 2000년도 춘계학술심포지엄
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• pp.1-3
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• 2000

A kind of traditional herbal prescription, Sip-Jeon-Dae-Bo-Tang (TJ-48), has been reported to improve the general condition of cancer patients receiving chemotherapy and /or radiation therapy, and to accelerate hematopoietic recovery from bone marrow injury by mitomycin C. In the present studies, we found that hot-water extract from Atractylodes lancea DC. rhizomes contributed mainly to intestinal immune modulating activity of TJ-48 on Peyer's patch cells mediated-hematopoietic response. After the fractionation, ALR-5 II a-1-1, 5 II b-2-2 and 5 II c-3-1 were further purified from crude polysaccharide fraction. Chemical analyses of each fraction indicated that ALR-5 II a-1-1 mainly contained arabinogalactan fraction whereas ALR-5 II b-2-2 and 5 II c-3-1 mostly comprised pectic polysaccharide fractions as the active polysaccharide ingredients. In order to analyze the essential structure of the activity, ALR-5 II a-1-1 was treated by sequential enzymatic digestion using exo-${\alpha}$-L-arabinofuranosidase and exo-${\beta}$-D-(1\longrightarrow3)-galactanase. Based upon the results of chemical and MALDI-TOF-MS analyses and activity on the digested fractions, the galactosyl side chains consisting of 6-linked Galf and Galp over tetrasaccharide in ALR-5 II a-1-1 might be responsible for the potent intestinal immune modulating activity. To characterize moiety of ALR-5 II c-3-1 for the expression of activity, endo-${\alpha}$-D-(1\longrightarrow4)-polygal acturonase (GL-PGase) purified from dried leaves of Panax ginseng digested ALR-5 II c-3-1. The results of structural analyses and activity on the digested fractions showed that PG-2, which structurally resembles to rhamnogalacturonan II (RG II), and PG-3 (galacturono-oligosaccharides) contained potent intestinal immune modulating activity. Further purification of the other acidic fraction (ALR-5 II b-2-2) indicated that ALR-5 II b-2-2Bb showed that the most potent activity. ALR-5 II b-2-2Bb also contained the unusual component sugars characteristics in RG- II as well as PG-2 derived from ALR-5 II c-3-1, but it could not be digested with GL-PGase. The present studies of relationship between structures and intestinal immune modulating activity of the active polysaccharides purified from A. lancea DC. rhizomes suggested that neutral galactosyl chains consisting mainly of (1\longrightarrow6)-linked Galf and Galp, and RG- II -like moiety with unique component sugars, such as 2-Me-Xyl, 2-Me-Fuc, Api, AceA, Kdo and Dha should play an important role in the potent intestinal immune modulating action of A. lancea DC. rhizomes.

• 대한한의학방제학회지
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• 제25권2호
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• pp.179-191
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• 2017

Objectives : Increased oxidative stress by reactive oxygen species (ROS) has been suggested as a major cause of muscle fatigue. Although several studies have demonstrated the various biological properties of Sophora flavescens Aiton, Glycyrrhiza uralensis Fischer and Dictamnus dasycarpus Turcz, but the antioxidative potentials have not been clearly demonstrated. The present study was designed to investigate the protective effects of their water and ethanol extract mixtures (medicinal herbal mixtures, MHMIXs) on hydrogen peroxide ($H_2O_2$)-induced cell damage and apoptosis in C2C12 myoblasts. Methods : Cytotoxicity was assessed by an MTT assay. Quantitative evaluation of apoptosis induction and ROS production was evaluated by flow cytometry analysis. Expression levels of apoptosis regulatory and DNA-damage proteins were detected by Western blotting. Result : The inhibition of $H_2O_2$-induced cell proliferation was effectively blocked in extracts of 3: 1: 1 (EMHMIXs-1) or 2: 2: 1 (EMHMIXs-2) of S. flavescens, G. uralensis and D. dasycarpus Turcz, ethanol extracts from various complex extracts in C2C12 myoblasts. EMHMIXs-1 and EMHMIXs-2 also effectively attenuated $H_2O_2$-induced C2C12 cell apoptosis, which was associated with the restoration of the upregulation of Bad and death receptor 4, and downregulation of XIAP and cIAP-1 induced by $H_2O_2$. In addition, these herbal mixtures significantly blocked the $H_2O_2$-induced ROS generation and phosphorylation of $p-{\gamma}H2A.X$, which suggests that they can prevent $H_2O_2$-induced cellular DNA damage. Conclusions : The results suggest that EMHMIXs-1 and EMHMIXs-2 could block the DAN damage and apoptosis of C2C12 myoblasts by oxidative stress through blocking ROS generation.