• Journal of Ginseng Research
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• 제45권1호
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• pp.134-148
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• 2021

Background: Lung cancer has a high incidence worldwide, and most lung cancer-associated deaths are attributable to cancer metastasis. Although several medicinal properties of Panax ginseng Meyer have been reported, the effect of ginsenosides Rk1 and Rg5 on epithelial-mesenchymal transition (EMT) stimulated by transforming growth factor beta 1 (TGF-β1) and self-renewal in A549 cells is relatively unknown. Methods: We treated TGF-β1 or alternatively Rk1 and Rg5 in A549 cells. We used western blot analysis, real-time polymerase chain reaction (qPCR), wound healing assay, Matrigel invasion assay, and anoikis assays to determine the effect of Rk1 and Rg5 on TGF-mediated EMT in lung cancer cell. In addition, we performed tumorsphere formation assays and real-time PCR to evaluate the stem-like properties. Results: EMT is induced by TGF-β1 in A549 cells causing the development of cancer stem-like features. Expression of E-cadherin, an epithelial marker, decreased and an increase in vimentin expression was noted. Cell mobility, invasiveness, and anoikis resistance were enhanced with TGF-β1 treatment. In addition, the expression of stem cell markers, CD44, and CD133, was also increased. Treatment with Rk1 and Rg5 suppressed EMT by TGF-β1 and the development of stemness in a dose-dependent manner. Additionally, Rk1 and Rg5 markedly suppressed TGF-β1-induced metalloproteinase-2/9 (MMP2/9) activity, and activation of Smad2/3 and nuclear factor kappa B/extra-cellular signal regulated kinases (NF-kB/ERK) pathways in lung cancer cells. Conclusions: Rk1 and Rg5 regulate the EMT inducing TGF-β1 by suppressing the Smad and NF-κB/ERK pathways (non-Smad pathway).

• 한방재활의학과학회지
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• 제24권3호
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• pp.51-69
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• 2014

Objectives This study was aimed to investigate the effects of Sibjeondaebotanggamibang (SJT) on the wound-induced rats. Methods It was observed the effects of anti-oxidation and anti-inflammation by using of lipopolysaccharide (LPS)-treated RAW 264.7 cells. For the observing on SJT anti-oxidation, it needed to mesure the total amount of polyphenol, DPPH scavenging ability, ABTS scavenging ability and the value of ROS production. In order to observe on the anti-inflammation of SJT, it was mesured the value of No and Cytokine (TNF-${\alpha}$, IL-$1{\beta}$, IL-6). It needed to make a scar (around $2{\times}2cm^2$) on the top of the fascia in the back of the rats and then the rats were divided into 4 groups (n=6). Control group was not treated at all, whereas SJ group was orally medicated SJT, Terra group was per-cutaneously applied Terramycin, and SJ+Terra group was both orally medicated SJT and percutaneously applied Terramycin per day for three weeks. The size of wound was measured with Digimatic Caliper and the blood samples (WBC, neutrophil, monocyte, lymphocyte) were analyzed using Minos-ST, which were collected by cardiac puncture. The effect on inflammatory cytokine (TNF-${\alpha}$, IL-$1{\beta}$, IL-6), immunological cells in synovial fluid was measured. To measure the wound factor expressed by wounded skin sample, we extracted RNA and to investigate MMP-1,2,9 we used RT-PCR. For performing histopathological examinations, we paralyzed the rats by ether, and extracted wounded skin tissues, which were measured by H & E, and monitored on the optical microscope. Results 1. DPPH and ABTS scavenging activity of SJT was increased concentration-dependantly, and ROS scavenging activity was significantly increased (10, $100{\mu}g/ml$). 2. NO production was significantly reduced in SJT treated cells ($100{\mu}g/ml$), both TNF-${\alpha}$ and IL-6 in SJT treated cells (1, 10, $100{\mu}g/ml$), and IL-$1{\beta}$ in SJT treated cells (1, $100{\mu}g/ml$). 1. The size of wound was significantly decreasing in SJ group, Terra group, SJ+Terra group. 2. WBC was significantly reduced in SJ and SJ+Terra group, monocyte in SJ+Terra group. Neutrophil was also reduced in SJ, SJ+Terra group but meaningless. 3. TNF-${\alpha}$ and IL-6 were significantly reduced in SJ group, Terra group, SJ+Terra group, and IL-$1{\beta}$ in SJ+Terra group. 4. mRNA expression in MMP-1 was significantly reduced in SJ group. 5. Collegan production and chronic inflammation were significantly decreased in SJ group, Terra group, SJ+Terra groups. Re-epithelization on the skin in Terra group, SJ+Terra groups was decreased. Conclusions According to this in vitro experiment, Sibjeondaebotanggamibang (SJT) has the effects of anti-oxidative and anti-inflammatory. By in vivo experiment, SJT has the effects of anti-inflammatory. Moreover, the progress of recovery was found visually, heamatologically, genetically and histopathologically. In conclusion, it could be thought that SJT has effect on the treating of wound.

• 식품과학과 산업
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• 제53권1호
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• pp.2-32
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• 2020

키토산은 천연에 존재하는 풍부한 고분자 다당류이며 동시에 항균작용, 항암작용, 면역 활성 증강 작용 등 다양한 생리 기능성을 가진 생체 물질(biomaterial)이다. 그러나 키토산은 수용액에 쉽게 용해되지 않고 체내 흡수율도 매우 낮은 고분자 물질이기 때문에 그 자체로는 생체 내에서 기능을 발휘하지 못한다. 따라서 체내 흡수율이 용이하고 보다 다양한 생리 활성을 나타내는 키토산올리고당으로 섭취할 필요가 있다. 키토산으로부터 키토산올리고당을 만드는 방법은 온화한 조건에서 부반응이 없는 효소분해가 가장 이상적이다. 그러나 키토산을 분해시킬 수 있는 효소의 가격이 매우 비싸고 효소 활성이 낮아 산업적으로 효소를 이용하여 키토산올리고당을 대량 생산하는데는 경제적으로 큰 문제가 있었다. 따라서 효소 분해에 의한 키토산올리고당의 생산을 산업적으로 적용시켰을 때 대량으로 소모되는 효소의 높은 생산 비용을 경감하기 위하여 효소의 재이용을 위한 생산 시스템을 도입하였고 한외여과막과 효소 반응기를 결합시킨 한외여과막 효소반응기를 키토산올리고당의 생산에 적용시킴으로써 대량 생산이 가능해졌다. 이 시스템은 사용하는 막의 종류에 따라 원하는 분자량의 키토산올리고당을 생산 할 수 있어 분자량 크기에 따라 생체 내에서 다른 생리기능을 나타내는 키토산올리고당을 각기 목적에 맞는 제품으로 대량 생산할 수 있다. 키토산올리고당은 암세포 성장에 영향을 미치는 면역을 증진시키고 암 전이 관련 효소인 MMP-2 및 MMP-9의 발현을 저지하여 암 전이를 억제할 뿐만 아니라 암세포의 신생 혈관형성을 억제하므로 암의 예방이나 치료에 활용될 수 있다. 또한 키토산올리고당은 생체 노화를 촉진시키는 초과산화 라디칼, 히드록시라디칼, 과산화 라디칼과 같은 활성산소종을 소거시킴으로써 활성산소종에 의해 유발되는 질환도 예방한다. 더 나아가서, 키토산올리고당은 양전하를 가진 아미노기를 갖고 있어 미생물 세포막의 음전하와 결합하여 세포막의 기능을 상실시킴으로써 막을 통해 출입하는 대사에 필요한 물질들을 봉쇄한다. 이것은 미생물의 성장과 증식을 감소시키는 효과가 있어 식품분야에서도 천연 보존제로서 활용될 것으로 기대된다. 지금까지 고혈압 및 심장 질환의 치료는 레닌엔지오텐신 시스템(RAS)경로의 치료조작(mani pulator) 및 ACE 저해에 초점이 맞추어져 왔다. Captopril, Enalapril, Alcacepril 및 Lisinopril같은 합성 ACE 저해제는 고혈압 치료 및 예방을 위해서도 널리 사용되고 있으나 기침, 알레르기 반응, 맛 장애 및 피부발진과 같은 부작용을 야기시킬 위험이 있다. 그러므로 고혈압의 치료나 관리를 위한 치료제로서 자연계에 존재하는 천연성분인 키토산올리고당이 바람직한 대안으로 사용될 수 있을 것이다. 고령 사회로 진입하면서 노인의 치매 발병률은 현저하게 증가하는 추세이나 아직까지 효과가 뛰어난 치매치료제는 개발되지 않고 있는 실정이다. 키토산올리고당이 치매유발효소인 베타-세크레테이즈뿐만 아니라 아세틸콜린 에스테르가수분해효소를 저해하고, 산화에 의한 뇌세포의 손상을 저해하는 효능이 있는 것으로 밝혀져 앞으로 치매 예방이나 치료에 적극적으로 활용되어야 한다. 최근에 소비자나 환자들은 다양한 부작용을 낳는 화학적으로 합성된 의약품을 기피하는 경향이 있어 자연식품이나 천연 생리기능성 물질과 자가치료에 대한 욕구가 높아지고 있다. 그러므로 의약품에 비해 다소 효능이 낮을지라도 특정한 질병의 예방이나 치료를 위해 특수한 생리 기능성 물질의 섭취는 더욱 더 인기를 끌게 될 것이다. 따라서 항암, 항산화, 항염증, 항균, 항고혈압, 항치매, 항당뇨, 항알레르기 등 다양한 생리활성을 나타내는 키토산올리고당을 활용한 건강기능성식품(nutraceuticals), 의약품(pharmaceuticals) 및 기능성 화장품(cosmeceuticals) 등 다양한 제품이 개발될 것으로 기대된다.

• Journal of Oral Medicine and Pain
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• 제32권3호
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• pp.251-261
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• 2007

담즙산과 합성담즙산유도체가 여러 종류의 암세포에 세포자멸사(apoptosis)를 유도하고 항암효과가 있다고 알려져 있다. 합성 chenodeoxycholic acid (CDCA) 유도체가 여러 가지 암세포에 유도한 세포자멸사 in vitro 연구들이 보고되어져 왔다. 하지만 아직 까지 구강편평상피암종세포에 합성 CDCA 유도체가 유도한 세포자멸사 연구는 없었다. 그래서 본 연구는 합성 CDCA 유도체인 HS-1199와 HS-1200이 사람구강편평상피암종세포에 세포자멸사 효과와 세포자멸사 기작을 알기 위해서 수행되었다. 합성 CDCA 유도체로 처리된 사람구강편평상피암종세포(YD9 세포)에서 caspase-3의 활성화, DFF의 degradation, poly (ADP-ribose) polymerase(PARP)의 분절화(HS-1200 only), DNA 분절화(HS-1200 only), 핵 응축, proteosome 활성화의 저해, 사립체막전위 (MMP)의 감소(HS-1200 only) 그리고 cytochrome c와 AIF의 사립체에서 세포질로의 유리와 같은 세포자멸사의 증거를 보였다. 그리고 두 개의 합성 CDCA 유도체 중에서 HS-1200이 HS-1199보다 더욱 더 강한 세포자멸사 효과를 보였다. 이 결과는 HS-1200이 YD9 세포에 항암효과를 가진다는 것을 증명한 것이다. 본 연구는 CDCA 유도체인 HS-1200이 사람구강편평상피암종세포에서 사립체 경로를 통한 caspase 의존적 세포자멸사를 강력하게 유도한다는 것을 증명했으며, 이러한 결과는 HS-1200이 사람구강편평상피암종의 치료적 전략으로서의 가능성이 높다고 생각한다.

• 대한한방내과학회지
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• 제23권2호
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• pp.165-173
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• 2002

Purpose : Among numerous biological symptoms of cancer, matrix metalloproteinases (MMPs) are essential for tumor invasion and metastasis. HAD is used as an inhibitor of MMP gene. This study was designed to evaluate the effects of HAD on anti metastasis and preventing recurrence in cancer patients. Materials and Methods : We retrospectively analyzed the medical records of 69 cancer patients who had been administered with HAD for over 12 months continuously in East-West Cancer Center of Oriental Hospital of Daejeon University, from January 1993 to May 2002. Results : We analyzed gender, portion, stage and anti-metastasis & recurrence rates of cancer patients. Analysis of sex cases showed that the percentage of male is 62.3%, female is 37.7%. Analysis of cancer portion showed that the percentage of stomach is 31.9%, colorectum is 26.1%, lung is 21.7%, liver is 8.7%, breast is 8.7% Analysis of stage showed that the rate of III is 78.3%, IV is 13.0% and II is 8.7%. Analysis of anti-metastasis and recurrence rates showed that colorectal cancer is 77.8%, stomach cancer is 63.6%, lung cancer is 33.4% and breast cancer is 33.3% (mean : 53.6%). Conclusions : HAD has significant effects on anti-metastasis and preventing recurrence of tumor on cancer patients. So it helps to prolong the survival rates of cancer patients.

• 대한암한의학회지
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• 제11권1호
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• pp.41-54
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• 2006

Ekongsan (EKS) was expected to have inhibitory effects on angiogenesis, considering the fact that its constituents such as Ginseng Radix, Glycyrrhizae Radix and Citri Pericarpium were reported to inhibit angiogenesis. Moreover, recently several metabolites transformed by the human intestinal microflora were reported to enhance effectiveness compared to their crude drugs. Based on these data, this study was designed to confirm whether the EKS metabolites (EKS-M) can significantly exert the anti-angiogenic and anti-metastatic activites. Hence, with EKS and EKS-M, viability assay, proliferation assay, in vitro tube formation assay, gelatin zymogram assay, in vitro invasion assay were carried out. EKS showed less toxicity in ECV304 and HT1080 cells than EKS-M. EKS-M inhibited the proliferation of HT1080 cells by 30% at 200 ${\mu}g/m{\ell}$ and 42% at 400 ${\mu}g/m{\ell}$ respectively. Also, EKS-M degraded the tube network at 200 ${\mu}g/m{\ell}$. EKS and EKS-M inhibited the expression of MMP-9 at 200 and 400 ${\mu}g/m{\ell}$in HT1080 cells. EKS reduced the invasive activity of HT1080 cells through matrigel coated transfilter at the concentration of 200 ${\mu}g/m{\ell}$ more effectively than EKS-M. These data suggest that EKS and EKS-M has anti-angiogenic and anti-metastatic activities.

• 대한암한의학회지
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• 제11권1호
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• pp.75-93
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• 2006

Jiaweicitaowan (JWCTW) has been used to inhibit recurrence and metastasis of cancer in clinical practice. Further study has shown its anti-metastatic and anti-angiogenic effects. By applying in vitro and in vivo anti-angiogenic evaluation model, the author assayed the each ingredients of JWCTW. The author performed the following experiments and the results are listed as below: Cell viability assay showed that the viability was almost identical between that of the control and that of the ingredients extract 40 ${\mu}g/m{\ell}$ treated, except of hexane fraction of Curcumae Radix (40 ${\mu}g/m{\ell}$, 2.0% of control), ethylacetate fraction (40 ${\mu}g/m{\ell}$, 26.7%), butanol fraction (20 ${\mu}g/m{\ell}$, 87.2%; 40 ${\mu}g/m{\ell}$, 12.5%) of Cremastrae appendiculatae Tuber, water fraction of Persicae Semen (40 ${\mu}g/m{\ell}$, 82.7%), ethylacetate fraction of Hippocampus (40 ${\mu}g/m{\ell}$, 85.3%). The results of gelatin zymogram assay showed that the ingredients of JWCTW decreases the gelatinolytic activity of MMP-9 from ECV304, at the concentration of 10 ${\mu}g/m{\ell}$. In in vitro invasion assay, the ingredients of JWCTW effectively inhibited the invasion of cancer cells as compared with the control (+PMA) groups. In capillary-like tube formation assay, the hexane and ethylacetate fractions of Curcumae Radix, Cremastrae appendiculatae Tuber and Persicae Semen showed the dramatic inhibition effects on tube formation of ECV304 at the concentration of 40 ${\mu}g/m{\ell}$. In ex vivo rat aortic ring assay, the hexane and ethylacetate fractions of Curcumae Radix, Cremastrae appendiculatae Tuber and Persicae Semen showed the inhibition effects on angiogenesis of rat aorta at the concentration of 40 ${\mu}g/m{\ell}$. According to the above research, the anti-angiogenic effects of the ingredients of JWCTW was proved and it suggested that the more effective prescription for anti angiogenesis could be developed.

• 대한암한의학회지
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• 제11권1호
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• pp.105-118
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• 2006

Two of the essential processes required for metastasis are neoangiogenesis and tumor cell invasion of basement membranes (BM) and extracellular matrix (ECM). Recently, data showed that herbs removing blood stasis has an anti-angiogenic effects. Tonifying vital Qi and eliminating pathogenic factor was a basic modality in Oriental oncology. In this study, we investigated several Qi and Blood tonics for potent angiogenic inhibitors. Methanol extracts of samples inhibited the proliferation of ECV-304 at the concentration of 100 ${\mu}g/m{\ell}$. Zizyphi Fructus, Glycyrrhizae Radix, Angelicae Gigantis Radix decreased the gelatinolytic activity of MMP-9 from ECV-304, at the concentration of 100 ${\mu}g/m{\ell}$ in gelatin zymography. In in vitro invasion assay, herbs inhibited the invasion activity of ECV-304 by 53% of control (Ginseng Radix), 39% (Zizyphi Fructus), 36% (Angelicae Gigantis Radix), 25% (Glycyrrhizae Radix). Ginseng Radix inhibited the capillary-like tube formation of ECV-304 at the concentration of 160 ${\mu}g/m{\ell}$, Angelicae Gigantis Radix and Paeoniae Radix Alba inhibited at the concentration of 320 ${\mu}g/m{\ell}$. These results indicated that Ginseng Radix, Glycyrrhizae Radix, and Angelicae Gigantis Radix could be considered as potent angiogenic inhibitiors.

• 대한암한의학회지
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• 제11권1호
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• pp.119-134
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• 2006

Shiquandabutang is very famous prescription for tonifying vital energy. We examined the anti-metatstastic effect of Shiquandabutang with in vitro invasion assay model. We performed the following experiments and the results are listed below:Cell viability assay was carried to determine the dose of Shiquandabutang. At lower dose under 200 ${\mu}g/m{\ell}$ (89.6%) viability was very high. But, viability downed as dose grows. At the dose of 600 ${\mu}g/m{\ell}$ (54.2%) viability was almost half of that of control. And at high dose of 1000 ${\mu}g/m{\ell}$ (15.8%) viability was very pure. In BrdU incorporation assay, Shiquandabutang treated groups showed the decreased DNA synthesis rate compared with control group.(200 ${\mu}g/m{\ell}$ (64.4%), 400 ${\mu}g/m{\ell}$ (7.3%)) The results of gelatinase assay showed that Shiquandabutang decreases the gelatinolytic activity of MMP-9. We examined tube formation assay and the result was that Shiquandabutang ihhibits the tube formation at the dose of 200 ${\mu}g/m{\ell}$ and 400 ${\mu}g/m{\ell}$. We examined rat aortic ring assay and the result was that Shiquandabutang ihhibits the angiogenesis of the rat aortic ring at the dose of 400 ${\mu}g/m{\ell}$. From our research, part of the mechanism underlying anti-metastastic effect of Shiquandabutang was proven in vitro. Moreover, we knew that Shiquandabutang is more effectively inhibits the angiogenesis at high dose.

• Journal of Acupuncture Research
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• 제29권1호
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• pp.75-87
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• 2012

Objectives : The purpose of this study is to inquire into the effect of different concentrations of bee venom pharmacopuncture to inhibit genesis of pro-inflammatory cytokines and to inhibit nuclear factor (NF)-${\kappa}B$ activation on type II collagen induced arthritis. Methods : The experiment was divided into category of the normal group (NOR)-no treated group, control group (CON)-CIA (collagen induced arthritis) induced group, and 4,000 : 1 bee venom group (BV-L)- 4000:1 bee venom pharmacopuncture treated group after CIA, and 2000:1 bee venom group (BV-H)- 2,000 : 1 Bee venom pharmacopuncture treated group after CIA. RA was induced in the mice via injecting $50{\mu}{\ell}$ C II mixed CFA. The bee venom pharmacopuncture was applied on $ST_{35}$ for 19 days from the 3rd day of RA inducement. To research the effect on the expression of IKK ($I{\kappa}B$ kinase), iNOS (inducible nitric oxide synthase) & COX-2 (cyclooxygenase-2) mRNA, RT-PCR was performed on synovial membrane cells from the knee joint of CIA mice. Results : The PMA-induced $I{\kappa}B$ kinase (IKK), inducible nitric oxide synthase (iNOS) and cyclooxygenase -2 (COX-2) mRNA expression were dose-dependantly decreased in bee venom treated with synoviocytes. In mice treated with bee venom pharmacopuncture, foot thickness and the damage of synovial membranes of the joint was lessened, and the activation of RA-related pro-inflammatory cytokines such as MIF, TNF-${\alpha}$ and MMP-9 was significantly decreased. The activation of iNOS and COX-2 was suppressed by the inhibition of NF-${\kappa}B$. In addition, each data was shown that 2,000 : 1 bee venom pharmacopuncture was more effective than 4,000 : 1 bee venom pharmacopuncture. Conclusions : It is speculated that bee venom pharmacopuncture has the therapeutic effect of palliating the damage of the synovial membrane and inflammation on RA by suppressing of NF-${\kappa}B$ activation.