• Journal of Nutrition and Health
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• 제42권5호
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• pp.434-441
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• 2009

본 연구는 J774 대식세포에서 FPN 유전자 발현 조절에 구리가 미치는 영향을 알아보기 위하여 수행되었으며 그 결과는 다음과 같다. J774 대식 세포에 구리를 처리하였을 때, iron exporter FPN의 mRNA 수준이 농도 의존적으로 증가하는 것으로 나타났다. 반면, iron importer DMT1의 mRNA 수준은 구리 처리에 의해 영향을 받지 않았다. Actinomycin D를 이용하여 mRNA 합성을 억제한 상태에서 FPN mRNA 분해 정도를 시간별로 추적한 결과, acitnomycin D 처리 후 9시간 경과시 FPN mRNA 수준이 처음 수준의 약 60% 정도로 감소하였다. 배양액에 구리를 첨가한 경우에도 FPN mRNA의 분해 정도는 아무것도 처리하지 않은 대조군과 유의적인 차이가 없었으며, 이로 볼 때 구리는 FPN mRNA의 안정성에 영향을 미치지 않는 것으로 생각된다. 한편, reporter assay 실험 결과 구리의 첨가는 FPN 프로모터 활성을 유의적으로 증가시키는 것으로 나타나, 구리가 FPN mRNA의 전사 과정을 직접적으로 촉진함을 알 수 있었다. 또한, FPN 5'-UTR에 위치하는 IRE (iron response element)의 존재 여부는 구리에 의한 FPN 전사 개시 활성에 영향을 주지 않는 것으로 나타났으며, 이로 볼 때 구리는 철분과는 독립적인 작용 기작에 의해 FPN 유전자 발현을 조절하는 것으로 사료된다. 이상의 결과를 종합해 볼 때, 구리는 대식 세포에서 전사개시 과정을 활성화함으로써 농도 의존적으로 FPN 유전자 발현을 촉진하는 것으로 생각되며, 이는 구리가 철분의 대사에 미치는 새로운 작용 기작을 제시한다. 앞으로, 구리와 철 분의 상호 작용이 FPN의 철분 및 다른 무기질 이온의 세포내 외 수송 (transport)에 어떤 영향을 미치는지에 대한 기능적 연구가 계속적으로 이루어져야 할 것으로 사료된다.

• 대한약리학회지
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• 제8권1호
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• pp.67-75
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• 1972

Many studies on the mechanism of the inotropic action of cardiac glycosides have shown the possible intimate relationship between the mobilization of intracellular calcium and inotropic effect. Evidence obtained from recent studies suggests that cardiac glycosides may increase the intracellular $Ca^{++}$ concentration through the release of this ion from cellular or intracellular membrane. It seemed imperative to study the effect of ouabain on the interaction between mitochondria and $Ca^{++}$, because mitochondria are known to have a rather powerful $Ca^{++}$ pump mechanism which may have an important role on the regulation of intracellular $Ca^{++}$ concentration. The present investigations was made into the effect of ouabain on $Ca^{++}$ untake of mitochondria in the presence of ATP and its dependence on $K^+$ and $Na^+$ in the medium. The results are summarized as follows: 1. The rate of rise in the turbidity of superprecipitation was solely influenced by ionic strength of the medium, not by the species of ion, i.e. $Na^+$ or $K^+$. The higher ionic strength suppressed and the lower enhanced the rate of superprecipitation respectively. 2. No effect of ouabain was found on the rate of superprecipitation. 3. Mitochondria depressed the rate of superpretipitation, and the depressed rate of superprecipitation by mitochondria was reversed by ouabain, and the degree of this reversal was almost identical in $Na^+$ and $K^+$ medium. 4. $Ca^{++}$ uptake of mitochondria was inhibited by ouabain in the presence of ATP and the degree of inhibition showed the dose-response manner in terms of concentration of ouabain. 5. In the absence of ATP, mitochondria took or the $Ca^{++}$ in initial period but released it later. Such uptake and release of $Ca^{++}$ was not influenced by ouabain. 6. It is suggested that intracellular calcium mobilization by ouabain through the action upon the mitochondria was due to inhibition on ATP-dependent $Ca^{++}$ uptake by this agent, not to the action upon so called binding.

• 한국산학기술학회논문지
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• 제17권11호
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• pp.640-647
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• 2016

형광 간섭 현상을 최소화시켜 상대적으로 민감한 측정이 가능한 aequorin기반 luminescence기술은 $G_{{\alpha}16}$ 단백질 도입을 통해 세포 내부의 칼슘 이동 신호를 감지하여 G 단백질 결합 수용체(G protein-coupled receptor, GPCR)의 기능 분석을 가능하게 하는 세포 기반 분석 기술로 수용체 및 G 단백질 유전자 전달의 최적화 과정이 필수적이다. 본 연구를 위해 corticotropin releasing factor receptor subtype 2(CRF2) 수용체를 모델 시스템으로 CRF2와 $G_{{\alpha}16}$ 단백질이 구축된 세 가지 안정화 세포주를 제작하였고, 이들을 이용한 서로 다른 세 가지 조건의 임시 발현 세포주에서 작용제(sauvagine)와 길항제(K41498)의 반응성을 분석하여 최적의 유전자 전달 방법을 도출하고자 하였다. 그 결과 sauvagine 및 K41498의 농도에 따른 반응에서 CRF2-$G_{{\alpha}16}$ 안정화 세포주가 임시 발현 세포주보다 10배 이상의 유효신호 비율을 나타내었고(z'=0.77) 임시 발현 세포주의 경우 $G_{{\alpha}16}$의 안정화 발현 이후에 CRF2를 전달하는 경우가 다른 임시 발현 조건보다 2배 이상 높은 효율을 보였다(z'=0.84). 따라서 임시 유전자 전달 기술을 GPCR 세포 기능 분석 시스템에 활용할 경우 $G_{{\alpha}16}$ 단백질에 대한 안정화 세포주를 우선적으로 구축하고, 목표하는 다양한 수용체들을 단계적으로 발현시키는 것이 최선의 방법이라 판단된다.

• 약학회지
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• 제41권3호
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• pp.389-398
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• 1997

Non-adenergic non-cholinergic (NANC) innervation on the circular muscle of the rabbit gastric body was investigated by observing the magnitudy of relaxations induced by the elec trical field stimulation (EFS). Strips were cut from the greater curvature of the gastric body and stimulated with 5s trains of 0.5 ms pulses at 1-20 Hz, 40 V. The EFS induced transient frequency-dependent contractons, followed by a slowly recovering relaxation ewpecially at higher frequency of the EFS. In the presence of atropine and guanethidine, the contractions were virtually abolished, while the frequency-dependent relaxations by the EFS remained unaffected. The magnitude of relaxations progressively decreased as the location of the strips gets closer to the bottom of the gastric body. The relaxations were ablished by tetrodotoxin, indicating that their orgin is the NANC nerve stimulation. NG-nitro-L-arginine (L-NNA, 10-$100{\mu}M$), the inhibitor of nitric oxide (NO)-synthase, caused a concentration-dependent inhibition of the NANC relaxations. The inhibitory effects of L-NNA were not affected gy the location of the strips and were reversed by L-arginine, the precursor of NO-biosynthesis. Hemoglobin (20-$60{\mu}M$), a NO scavenger, inhibited the NANC relaxation s in a concentration-dependent manner. This inhibition was more prominent in the NANC relaxations observed in the lower portion of the gastric body and the relaxations induced ly lower frequencies of the EFS. Methyelne blue (10-$100{\mu}M$), an inhibitor of cytosolic guanylate cyclase, markedly inhibited the NANC relaxations, almost abolishing the response at a higher dose ($100{\mu}M$). These results suggest that NANX innervation of the rabbit gastric body progeressively decrease as he location of the strips gets closer to the bottom of the gastric body, and that the NANC relaxation is primarily mediated by NO-guanosine 3',5'-cyclic monophophate (cyclic GMP).

• 약학회지
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• 제48권6호
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• pp.328-334
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• 2004

Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human pa pilloma virus) type 16 and cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Mentha arvensis Linne var.piperascens have inhibitory effects on bindings between E6 oncoprotein and tumor suppressor p53, E3 ubiqutin- protein ligase (E6AP). HPV oncoprotein inhibitors from Mentha piperita L. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and inhibitory compounds were finally purified by HPLC using an ELISA screening system based on binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on functions of E6 oncoprotein. We investigated whether caffeic acid methyl ester (CAM) extracted from Mentha piperita L. could inhibit the function of E6 oncoprotein. CAM inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that CAM inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.

• Journal of Ginseng Research
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• 제24권1호
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• pp.18-22
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• 2000

앞의 연구에서 우리는 진세노사이드가 신경세포에 존재하는 high-threshold voltage-dependent $Ca^{2+}$ channel을 억제한다는 것을 발표하였다. 그러나, 이러한 연구는 진세노사이드가 여러 칼슘 채널subtypes중 어느 특정 칼슘 채널만을 선택적으로 조절한다는 것을 보여주지는 않았다. 따라서 이 연구에서 우리는 여러 칼슘 채널subtypes에 선택적으로 작용하는 약물 혹은 toxins을 이용하여 진세노사이드가 어느 종류의 칼슘 채널 subtypes를 억제하는가를 bovine chromaffin cell을 이용하여 연구하였다. 사용한 물질은nimodipine(L-type 칼슘 채널 길항제), $\omega$-conotoxin GVIA (N-type $Ca^{2+}$ channel 길항제), $\omega$-agatoxin IVA(P-type 칼슘 채널 길항제)이었다. 연구 결과 진세노사이드는 bovine chromaffin 세포에 존재하는 high-threshold 칼슘 current을 투여 농도별로 억제하였다. $IC_{50}$/은 약 120 $\mu$g/ml인 것으로 나타났다. nimodipine은 진세노사이드에 의한 칼슘 currents억제 작용에 영향을 미치지 않은 것으로 나타났다. 그러나, $\omega$-conotoxin GVIA, $\omega$-agatoxin IVA 및 nimodipine+$\omega$-conotoxin GVIA+$\omega$-agatoxin IVA을 처리한 세포에서는 진세노사이드에 의한 칼슘 currents억제 작용이 현저하게 줄어 들었다. 이러한 연구 결과들은 진세노사이드가 L-type 칼슘 채널은 억제하지 않고, 주로 N-, p-, 및 Q-type칼슘 채널을 억제한다는 것을 보여주고 있다

• 한국자원식물학회지
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• 제23권5호
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• pp.399-405
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• 2010

현재 임상적으로 적용되고 있는 항암요법들은 많은 부작용을 가지고 있으며, 부작용에 의하여 또다른 질환을 야기하는 것이 항암치료에서 문제점으로 지적되고 있다. 따라서 부작용을 줄이고 항암요법을 유지시키는 방법으로, 인체 안전하다고 보고된 천연물을 이용하여 면역력을 증가시킴으로써 인체내 항암 효과를 나타내게 하는 BRM들을 개발하는 연구가 큰 의미를 지닌다. 이에 본 연구에서는 민간요법으로 이미 사용되고 있는 해당화의 macrophage의 활성화에 대한 BRM 효과를 확인하였으며, 특히 과육(RRF)과 종자(RRS)의 부위별 추출물로 그 효과를 비교 분석하였다. RRF는 암세포 자체에 대한 세포독성은 나타내지 않았으나, macrophage의 활성화에 의한 항암 효과를 나타내었다. 이러한 항암 효과는 macrophage의 활성화에 의한 증가되는 NO 및 TNF-$\alpha$와 같은 암세포 독성물질에 의한 효과를 기대할 수 있으나, RRF 처리에 의하여 활성화된 macrophage는 NO 분비에 효과를 나타내지 않았다. RRF의 처리는 TNF-$\alpha$ 분비를 증가시켰으나, macrophage의 활성화에 의해 암세포 독성을 나타내지 않았던 RRS에서도 TNF-$\alpha$ 분비가 증가한 것으로 보아 TNF-$\alpha$ 분비만으로는 macrophage의 항암효과에 직접적인 영향을 나타내지는 않는 것으로 보인다. 본 연구 결과들은 종합해 볼 때, RRF는 RRS와는 달리 macrophage를 활성화하여 항암효과를 나타내었으며, phagocytosis 능력도 증가시켰다. RRF는 TNF-$\alpha$ 등의 분비조절과 같이 RRS와는 다르게 macrophage를 활성화시키는 것으로 보이며, 임상적으로 적용시 RRF가 RRS보다 세포독성 측면에서도 안전하고, macrophage의 활성화 효과 측면에서도 유의성이 높을 것으로 평가 된다.

• Journal of Acupuncture Research
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• 제29권1호
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• pp.115-125
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• 2012

Objectives : The purpose of this study is to observe the inhibitory effects of $Chrthami$ semen oil pharmacopuncture(CSOP) on CIA (collagen-induced arthritis) mice. Materials and Methods : Two types of experiments were conducted: $in$ $vitro$ assay, inhibition of MIF mRNA and TNF-${\alpha}$ mRNA expressions in synovial membranes was observed, and $in$ $vivo$ assay, $1{\mu}{\ell}/kg$ CSOP was injected every day to the left $Weizhong$ ($BL_{40}$) from day 3 to 21 after induction of CIA, and changes in paw edema, apical surface morphology, neovascularization in synovial membranes, fibrosis, pro-inflammatory cytokines production, Th-1 differentiation, and anti-inflammatory effect were investigated. Results : 1. In synoviocytes of the CIA mice treated with CSOP, MIF mRNA and TNF-${\alpha}$ mRNA expressions were down-regulated in a dose-dependent manner. 2. Paw edema of the CIA mice treated with CSOP was diminished. 3. Tissue injury in the synovial membranes, capillary distribution and fibrosis were reduced in CSOP-treated mice. 4. MIF, TNF-${\alpha}$, IL-6, MMP-9 expressions were repressed in CSOP-treated mice during the experiment to observe the inhibitory effect on cytokines production in early stage RA. 5. IL-12 and CD28 were reduced in CSOP-treated mice during the observation of inhibitory effect on Th 1 differentiation. 6. PPAR-${\gamma}$ was increased during the experiment to observe the anti-inflammatory effect of CSOP. Conclusions : The results may suggest that administration treatment using $Chrthami$ semen oil pharmacopuncture decreases the inflammatory response on an Animal Model with CIA.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제28권2호
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• pp.111-117
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• 2006

An area of current research is investigating the app1ication of human mesenchymal stem cells or hMSCs as a cell-based regenerative therapy. In order to achieve effective bone regeneration, appropriate matrices functioning as cell-carriers must be identified and optimized in terms of function, efficacy and biocompatibility. Two methods of approaching optimization of matrices are to facilitate adhesion of the donor hMSCs and furthermore to facilitate recruitment of host progenitor cells to osteoblastic differentiation. Pleiotrophin is an extracellular matrix protein that was first identified in developing rat brains and believed to be associated with developing neuronal pathways. A recent publication by Imai and colleagues demonstrated that transgenic mice with upregulated pleiotrophin expression developed a greater volume of cortical as well as cancellous bone. The proposed mechanism of action of pleiotrophin is demonstrated here. Through either environmental stresses and/or intracellular regulation, there is an increase in pleiotrophin production. The pleiotrophin is released extracellularly into areas requiring bone deposition. A receptor-mediated process recruits host osteoprogenitor cells into these areas. Therefore, the aim of our study was to investigate the osteoconductive properties of pleiotrophin. We wanted to determine if pleiotrophin coating facilitates cellular adhesion and furthermore if this has any effect on hMSCs derived bone formation in an animal model. The results showed a dose dependent response of cellular adhesion in fibronectin samples, and cellular adhesion was facilitated with increasing pleiotrophin concentrations. Histologic findings taken after 5 weeks implantation in SCID mouse showed no presence of bone formation with only a dense fibrous connective tissue. Possible explanations for the results of the osteogenesis assay include inappropriate cell loading.

• Asian Pacific Journal of Cancer Prevention
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• 제16권11호
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• pp.4781-4786
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• 2015

Siberian ginseng (Eleutherococcus senticosus) is used primarily as an adaptogen herb and also for its immune stimulant properties in Western herbal medicine. Another closely related species used in East Asian medicine systems i.e. Kampo, TCM (Manchuria, Korea, Japan and Ainu of Hokkaido) and also called Siberian ginseng (Acanthopanax senticosus) also displays immune-stimulant and anti-cancer properties. These may affect tumour growth and also provide an anti-fatigue effect for cancer patients, in particular for those suffering from lung cancer. There is some evidence that a carbohydrate in Siberian ginseng may possess not only immune stimulatory but also anti-tumour effects and also display other various anti-cancer properties. Our study aimed to determine the inhibitory and also proliferative effects of a methanol plant extract of Siberan ginseng (E. senticosus) on various cancer and normal cell lines including: A-549 (small cell lung cancer), XWLC-05 (Yunnan lung cancer cell line), CNE (human nasopharyngeal carcinoma cell line), HCT-116 (human colon cancer) and Beas-2b (human lung epithelial). These cell lines were treated with an extract from E. senticosus that was evaporated and reconstituted in DMSO. Treatment of A-549 (small cell lung cancer) cells with E. senticosus methanolic extract showed a concentration-dependent inhibitory trend from $12.5-50{\mu}g/mL$, and then a plateau, whereas at 12.5 and $25{\mu}g/mL$, there is a slight growth suppression in QBC-939 cells, but then a steady suppression from 50, 100 and $200{\mu}g/mL$. Further, in XWLC-05 (Yunnan lung cancer cell line), E. senticosus methanolic extract displayed an inhibitory effect which plateaued with increasing dosage. Next, in CNE (human nasopharyngeal carcinoma cell line) there was a dose dependent proliferative response, whereas in Beas-2 (human lung epithelial cell line), an inhibitory effect. Finally in colon cancer cell line (HCT-116) we observed an initially weak inhibitory effect and then plateau.