• Archives of Pharmacal Research
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• 제24권2호
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• pp.126-135
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• 2001

Quinacrine (QU), a phospholipase-A2 (PLA-2) inhibitor has been used clinically as a chemotherapeutic adjuvant. To understand the mechanisms leading to its chemotherapeutic effect, we have investigated QU-induced apoptotic signaling pathways in human cervical squamous carcinoma HeLa cells. In this study, we found that QU induced cytochrome c-dependent apoptotic signaling. The release of pro-apoptotic cytochrome c was QU concentration- and time-dependent, and preceded activation of caspase-9 and -3. Flow cytometric FACScan analysis using fluorescence intensities of $DiOC_6$/ demonstrated that QU-induced cytochrome c release was independent of mitochondrial permeability transition (MPT), since the concentrations of QU that induced cytochrome c release did not alter mitochondrial membrane potential (${\blacktriangle}{\Psi}_m$). Moreover, kinetic analysis of caspase activities showed that cytochrome c release led to the activation of caspase-9 and downstream death effector caspase-3, Caspase-3 inhibitor (Ac-DEVD-CHO) partially blocked QU-induced apoptosis, suggesting the importance of caspase-3 in this apoptotic signaling mechanism. Supplementation with arachidonic acid (AA) sustained caspase-3 activation induced by QU. Using inhibitors against cellular arachidonate metabolism of lipooxygenase (Nordihydroxyguaiaretic Acid, NDGA) and cyclooxygenase (5,8,11,14-Eicosatetraynoic Acid, ETYA) demonstrated that QU-induced apoptotic signaling may be dependent on its role as a PLA-2 inhibitor. Interestingly, NDCA attenuated QU-induced cytochrome c release, caspase activity as well as apoptotic cell death. The blockade of cytochrome c release by NDCA was much more effective than that attained with cyclosporin A (CsA), a MPT inhibitor. ETYA was not effective in blocking cytochrome c release, except under very high concentrations. Caspase inhibitor z-VAD blocked the release of cytochrome c suggesting that this signaling event is caspase dependent, and caspase-8 activation may be upstream of the mitochondrial events. In summary, we report that QU induced cytochrome c-dependent apoptotic signaling cascade, which may be dependent on its role as a PLA-2 inhibitor. This apoptotic mechanism induced by QU may contribute to its known chemotherapeutic effects.

• Archives of Pharmacal Research
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• 제22권5호
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• pp.485-490
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• 1999

The mechanism of action of fish oil (FO), currently used in different chronic inflammatory conditions such as rheumatoid arthritis (RA), is not completely understood, although it is thought that it could alter the metabolism of endogenous autacoids. In addition, we hypothesized that the known capability of fatty acids (FA) of stabilizing serum albumin and perhaps other proteins, may be of pharmacological relevance considering that it is shared by other anti-rheumatic agents (e.g. nonsteroidal antiinflammatory drugs). Thus, we studied the effect of oral administration of FO and corn oil (CO), a vegetable oil with a different composition, on the stability of rat serum proteins, evaluated buy a classical in vitro method based on heat-induced protein denaturation. FO, and, to a lower extent, CO inhibited heat-induced denaturation of rat serum (RS): based on the inhibitory activity (EC50) of the major fatty acids against heat-induced denaturation of RS in vitro, it was possible to speculate the in vivo effects of palmitic acid (C16:0) and eicosapentaenoic acid (EPA, C20:5, n-3) may be more relevant than that of linolenic acid (C18:2). To better investigate this phenomenon, we extracted albumin from the serum of animals treated or not with FO with a one-step affinity chromatography technique, obtaining high purity rat serum albumin preparations (RSA-CTRL and RSA-FO), as judged by SDS-PAGE with Coomassie blue staining. When these RSA preparations were heated at $70^{\circ}C$ for 30 min, it was noted that RSA-FO was much more stable than RSA-CTRL, presumably due to higher number of long chain fatty acids (FA) such as palmitic acid or EPA. In conclusion, we provided evidences that oral administration of FO in the rat stabilizes serum albumin, due to an increase in the number of protein bound long chain fatty acids (e.g. palitic acid and EPA). We speculate that the stabilization of serum albumin and perhaps other proteins could prevent changes of antigenicity due to protein denaturation and glycosylation, which may trigger pathological autoimmune responses, suggesting that this action may be involved in the mode of action of FO in RA and other chronic inflammatory diseases.

• 한국식품저장유통학회지
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• 제22권6호
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• pp.867-877
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• 2015

본 연구에서는 삼채 뿌리를 물, 에탄올, 메탄올로 추출했다. 추출방법에 따라 추출물의 항산화 및 항염증 효과는 다르게 확인됐다. 그 중에는 에탄올 추출물은 가장 좋은 효과를 확인하였으며 강한 환원력, DPPH 라디칼, superoxide 라디칼 및 아질산염 소거능이 나타났다 (IC50값은 각각 3.22, 4.26, 0.95, 3.22 mg/mL). 물 추출물과 메탄올 추출물은 좀 약한 효과를 나타났는데 좋은 hydroxyl 라디칼과 superoxide 라디칼 소거능을 볼 수 있다. RAW 264.7세포를 통해서 측정한 결과에 따라 모두 추출물은 유익적인 염증 개성 효과를 나타냈다. 에탄올 추출물은 가장 높은 NO 생성 저해 활성을 나타내고 그 다음은 물 추출물과 메탄올 추출물을 순으로 나타냈다. Total phenolic, flavonoid하고 thiosulfinate의 함량은 다 에탄올 추출물은 제일 높은 농도를 가지고 있다 (각각 24.96 mg GAE/g extract, 4.27 mg RE/g extract, $14.2{\mu}M/g$ extract). 연구결과에 따라 삼채 뿌리는 높은 total phenolic, total flavonoid하고 total thiosulfinate 함량을 함유하고 좋은 항산화 및 항염증 효과를 볼 수 있다.

• 생명과학회지
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• 제25권3호
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• pp.269-275
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• 2015

생체에서 가장 많은 비율로 분포하고 있는 콜라겐 펩타이드는 동물의 뼈와 해양 생물의 비늘에 많이 함유되어 있다. 콜라겐은 동물 생체의 여러 결합조직에서 구조 단백질로 흔하게 발견된다. 또한, 이들은 생물 의료 자재, 제약, 화장품, 식품 및 가죽 산업에 널리 이용된다. 각종 어류 비늘에서 추출된 펩타이드는 UVB 조사에 의해 유도된 피부의 손상 및 광 노화에 대한 보호 효과가 있었다. 그럼에도 불구하고, UVB 조사에 대한 옥돔 비늘 유래의 펩타이드 특성은 명확히 알려져 있지 않다. 이 연구에서는 옥돔 비늘 추출물에서 분리된 1 kDa 이상(HMP)과 1 kDa 이하(LMP)의 펩타이드를 이용하여 UVB 조사에 의해 유도된 피부 손상과 광 노화에 대한 효과를 연구하였다. 이들 펩타이드는 농도 의존적으로 DPPH 라디칼 소거능을 보였으며 LMP는 HaCaT 인간 피부세포에서 UVB 조사에 의해 유도된 세포 지질 과산화 산물인 8-isoprostane 생성을 억제하였다. 그리고 LMP와 HMP는 B16F10 마우스 흑색종 세포에서 tyrosinase 활성 및 melanin 함량을 감소시켰으며 또한 HaCaT 세포에서 UVB로 유도된 elastase 활성을 감소시켰고 matrix metalloproteinase-1의 활성을 감소시켰다. 이러한 결과는 옥돔 비늘에서 유래된 펩타이드가 미백효과, 항산화제 및 광 노화 억제제로서 유용한 물질이 될 것으로 기대된다.

• 생명과학회지
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• 제17권7호통권87호
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• pp.910-914
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• 2007

본 연구는 MRI 조영제나 새로운 생리활성 물질을 개발키 위해 paraformaldehyde와 hypophosphorous acid를 아미노산인 glycine 혹은 glutamic acid와 함께 반응시켜 3,7-dihydroxy-3,7-dioxoperhydro-1,5,3,7-diazadiphos-phocme-1,5-diacetic acid 1 와 3,7-dihydrexy-3,7-dioxoperhydre-1,5,3,7-diazadiphos-phocme-1,5-di- (2-glutaric acid)3을 합성하였다. 그러나 aspartic acid에 의한 2-[5-(1,2-dicarboxy-ethyl)-3,7-dihydroxy-3,7-dioxo-315.715-[1,5,3,7]diazadiphosphocan-1-yl]-succinic acid 2는 얻지 못했다. 합성 된 3,7-dihydroxy-3,7-dioxoperhydro-1,5,3,7-dia-zadiphosphocine-1,5-diacetic acid 1을 산 촉매에 의한 에스테르화반응시켜 화합물 3,7-dihydroxy-3,7-dioxoper-hydro-1,5,3,7-diazadiphosphocine-1,5-diacetic acid methyl ester 4, 3,7-dihydroxy-3,7-dioxoperhydro-1,5,3,7-dia-zadiphosphocine-1,5-diacetic acid ethyl ester 5, 그리고 3,7-dihyoxy-3,74dioxoperhyo-1,5,3,7-diazadiphosphocine-1,5-diacetic acid propyl ester 6을 합성하였다. 계속해서 다양한 반응을 통해 새로운 화합물을 합성할 것이며 생리활성검색도 실시할 예정이다.

• 생명과학회지
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• 제30권11호
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• pp.990-998
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• 2020

본 연구는 C57BL/6 마우스에서 Allium cepa (red) 생물전환 추출물과 Angelica gigas Nakai 추출물이 혼합된 새로운 물질의 모발 성장 촉진 효과를 평가하기 위해 수행되었다. 적양파의 에탄올 추출물을 Bacillus subtilis KJ-3(BS3) 균주의 사용을 통해 생물전환하였고 이것을 Red-BCQ라고 명명하였다. Red-BCQ의 퀘르세틴 함량은 생물전환 후 약 7.4배 증가 되었다. 참당귀 추출물(Agnex)에는 decursin (D), decursinol angelate (DA) 등 쿠마린이 다량 함유되어 있다. Agnex 1 mg 중에는 D 0.4146 mg, DA 0.3659 mg이 포함되어 있었다. 미녹시딜(minoxidil)은 모발 성장을 촉진하는 것으로 알려져 있다. 본 연구에서는 Red-BCQ, Agnex 및 이들 혼합물의 모발 성장 촉진효과를 5% 미녹시딜과 비교하였다. 25마리의 마우스는 생리식염수(CON), 5% 미녹시딜(PCON), Red-BCQ (RA), Agnex (AG), 이들 혼합물 (RAG) 처리군 등 5개의 실험군으로 나누었다. 시료는 하루에 한 번 4주 동안 정해진 시간에 경구로 투여되었다. 모발 증식은 7, 14, 21, 28일에 사진으로 관찰하였다. 또한 5α-reductase, alkaline phosphatase (ALP), γ-glutamyl transpeptidase (γ-GT), insulin-like growth factor (IGF)-1, transforming growth factor(TGF)-β1, 항산화효소활성, 피부조직의 모낭도 관찰했다. 모든 결과에서 혼합물 투여 그룹은 다른 그룹보다 항산화 효과와 모발 성장 촉진 효과가 더 컸다. 이러한 데이터는 RAG가 C57BL/6 종의 모발 성장에 대해 강력한 자극활동을 가지고 있음을 시사한다.

• Molecules and Cells
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• 제37권6호
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• pp.449-456
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• 2014

The use of synovial fluid-derived mesenchymal stem cells (SFMSCs) obtained from patients with degenerative arthropathy may serve as an alternative therapeutic strategy in osteoarthritis (OA) and rheumatoid arthritis (RA). For treatment of OA and RA patients, autologous transplantation of differentiated MSCs has several beneficial effects for cartilage regeneration including immunomodulatory activity. In this study, we induced chondrogenic differentiation of SFMSCs by inhibiting protein kinase A (PKA) with a small molecule and microRNA (miRNA). Chondrogenic differentiation was confirmed by PCR and immunocytochemistry using probes specific for aggrecan, the major cartilaginous proteoglycan gene. Absorbance of alcian blue stain to detect chondrogenic differentiation was increased in H-89 and/or miRNA-23b-transfected cells. Furthermore, expression of matrix metalloproteinase (MMP)-9 and MMP-2 was decreased in treated1 cells. Therefore, differentiation of SFMSCs into chondrocytes through inhibition of PKA signaling may be a therapeutic option for OA or RA patients.

• 한국자원식물학회지
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• 제22권5호
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• pp.431-437
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• 2009

in vivo 및 in vitro에서 마황의 항염증효과를 검토하기 위하여 마황추출물을 급여한 흰쥐에게 LPS shock로 급성기 염증반응을 유발시킨 후, 혈액 및 간장의 전염증성 cytokines들의 농도를 경시적으로 조사하였으며, 한편으로는 Raw 264.7 cell에 LPS shock를 가한 후, 마황추출물이 각종 전염증성 cytokines들의 생산량에 미치는 영향을 조사했다. in vivo 실험에서, 각 처리군 별 plasma IL-$1{\beta}$, IL-6, TNF-$\alpha$ 및 IL-10 농도의 경시적 변동은 전 처리군 모두가 LPS 처리 후, 2h째 급격하게 증가하여, 5h째에 최고치를 나타내었다. Plasma IL-$1{\beta}$, IL-6 및 TNF-$\alpha$의 농도는 LPS처리 후 5h째에서 마황 첨가군 모두가 대조군보다 낮은 값을 나타내었다. Plasma IL-10의 농도는 LPS처리 후, 2h째 및 5h째 모두 에서 마황추출물 첨가군 들이 대조군보다 높은 값을 나타내었다. Raw 264.7 macrophages를 이용한 in vitro 실험에서, IL-$1{\beta}$, IL-6 및 TNF-$\alpha$의 농도들은 대조군보다 마황처리 군들 모두가 낮은 값을 나타내었다. IL-10의 농도는 대조군보다 마황처리군들이 다소 높은 경향을 보였으나, 유의한 차이는 아니었다. 이상의 결과들을 종합해보면 마황에 내재하는 기능성 물질들이 염증반응을 완화하는 효과를 가지고 있음을 시사해 준다.

• Food Science and Biotechnology
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• 제15권1호
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• pp.63-69
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• 2006

The primary objective of this study is to determine whether a diet supplemented with brown rice koji (BRK) results in a reduced stress response in rats and mice. BRK, which has been suggested as a candidate for use as a stress- and fatigue-fighting supplement, was compared with red ginseng extract (RG) for its stress-reducing potential. The animals in this study were divided into no-stress, stress, RG, and BRK groups of 8 to 10 animals each. Stress was induced by means of immobilization (being restrained in plastic tubes for 30 min and electroshock (0.5 mA in mice or 2 mA in rats for 5 min). The no-stress group was not exposed to stress. Rats in the RG group received oral doses of 200 mg RG extract/kg body weight daily. The BRK group was fed a 30% BRK diet and exposed to stress. Animals were given supplements for 7 days before being exposed to stress, and then were given supplements for 5 days with exposure to stress. When the stress exposure ended, the animals were observed for stress-related changes in behavior and their plasma corticosterone levels were measured. BRK supplementation was associated with a partial blockade of the effects of stress on locomotion and elevated plus-maze test results in rats and mice. It was also associated with a partial reduction in stress-induced behaviors such as freezing, burrowing, smelling, face-washing, and rearing. BRK supplementation did not have a significant effect on plasma corticosterone levels, which were increased in the animals exposed to stress (p<0.01). The mice in the RG group received RG in water (2 mg RG/ mL $H_2O$), and the BRK group received a 30% BRK diet (weight) for 7 days. Both groups were evaluated for signs of fatigue. BRK supplementation increased endurance, as indicated by time on the rota-rod, in cold water, and on the horizontal wire. These results suggest that BRK supplementation partially protects the animal from the effects of stress and may also contribute to resistance to fatigue on physical exertion.

• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of International Convention of the Pharmaceutical Society of Korea
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• pp.109-113
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• 2001

Oxidized low-density lipoprotein (oxLDL) has been shown to modulate transactivations by the peroxisome proliferator activated receptor (PPAR)$\gamma$ and nuclear factor-kappa B (NF$\kappa$B). In this study, the oxLDL signaling pathways involved with the NF$\kappa$B transactivation were investigated by utilizing a reporter construct driven by three upstream NF$\kappa$B binding sites, and various pharmacological inhibitors. OxLDL and its constituent lysophophatidylcholine (lysoPC) induced a rapid and transient increase of intracellular calcium and stimulated the NF-KB transactivation in resting RAW264.7 macrophage cells in an oxidation-dependent manner. The NF$\kappa$B activation by oxLDL or lysoPC was inhibited by protein kinase C inhibitors or an intracellular calcium chelator. Tyrosine kinase or PI3 kinase inhibitors did not block the NF$\kappa$B transactivation. Furthermore, the oxLDL-induced NF$\kappa$B activity was abolished by the PPAR$\gamma$ ligands. When the endocytosis of oxLDL was blocked by cytochalasin B, the NF$\kappa$B transactivation by oxLDL was synergistically increased, while PPAR transactivation was blocked. These results suggest that oxLDL activates NF-$\kappa$B in resting macrophages via protein kinase C- and/or calcium-dependent pathways, which does not involve the endocytic processing of oxLDL. The endocytosis-dependent PPAR$\gamma$ activation by oxLDL may function as an inactivation route of the oxLDL induced NF$\kappa$B signal. Short heterodimer partner (SHP), specifically expressed in liver and a limited number of other tissues, is an unusual orphan nuclear receptor that lacks the conventional DNA-binding domain. In this work, we found that SHP expression is abundant in murine macrophage cell line RAW 264.7 but suppressed by oxLDL and its constituent I3-HODE, a ligand for peroxisome proliferator-activated receptor y. Furthermore, SHP acted as a transcription coactivator of nuclear factor-$\kappa$B (NF$\kappa$B) and was essential for the previously described NF$\kappa$B transactivation by lysoPC, one of the oxLDL constituents. Accordingly, NF$\kappa$B, transcriptionally active in the beginning, became progressively inert in oxLDL-treated RAW 264.7 cells, as oxLDL decreased the SHP expression. Thus, SHP appears to be an important modulatory component to regulate the transcriptional activities of NF$\kappa$B in oxLDL-treated, resting macrophage cells.