• 생명과학회지
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• 제20권9호
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• pp.1365-1372
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• 2010

본 연구에서는 마를 건조 후 분말화하여 실험에 사용하였으며, 건조 마 분말에 대한 생리활성에 대한 연구로 건조된 마를 유기용매로 추출하여 마 추출물과 분획물들의 $H_2O_2$에 의해 유도된 산화적 스트레스 및 인체 암세포(HT1080 인체 섬유육종세포, HT-29 인체 결장암세포)에 대한 증식 억제 효과에 대해 검토하고자 하였다. 마는 항비만, 항변비, 항돌연변이, 혈당 및 혈중 콜레스테롤 감소 등의 활성이 있어 건강식품으로 알려져 있다. $H_2O_2$에 의해 유도된 산화적 스트레스 저해효과를 알아보기 위하여 DCFH-DA assay를 행하였다. 건조 마 A+M 추출물을 농도별로 인체 섬유육종세포(HT1080)에 처리하였을 때 농도 의존적으로 세포 내 활성산소종을 크게 억제시켰다. 각 분획물들 중 85% aq. MeOH 분획물은 다른 분획물과 비교하여 높은 지질 과산화물 생성 억제 효과를 보였다. 인체 암세포에 대한 증식 억제 실험에서 A+M 및 MeOH 추출물은 농도 의존적으로 인체 섬유육종 및 결장암세포의 증식을 억제하였다. 마 추출물로부터 얻어진 n-hexane, 85% aq. MeOH 및 n-butanol (n-BuOH) 분획물들은 첨가농도 0.5 mg/ml 이상 처리했을 때 이들 두 암세포의 증식을 유의적으로 억제하였다(p<0.05). 본 연구 결과로 부터 건조 마의 암세포 증식 억제 효과는 마의 A+M 추출물이 MeOH 추출물에 비해 높았고, 분획물들의 경우에는 water 분획물을 제외한 분획물들의 암세포 억제 효과가 높았다. 특히 85% aq. MeOH 분획물은 인체 암세포 증식 억제 효과뿐만 아니라 세포 내 활성산소종 감소시키는 효과가 높아 이들 분획물에 활성물질이 있을 것으로 추정된다.

• 생명과학회지
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• 제28권3호
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• pp.293-299
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• 2018

본 연구는 PMA (Phorbol 12-myristate 13-acetate)에 의해 MMP 활성이 증가된 섬유육종세포에서 MMP-2와 -9의 mRNA 발현 및 단백질 활성에 대한 만형자의 억제 효과를 zymography와 RT-PCR 방법에 의해서 측정하였다. 만형자 시료는 dichloromethane에 의해서 두 번 추출되었으며 동일한 과정을 methanol를 사용하여 반복하였다. 각각의 용매에 의해서 얻어진 추출물을 합한 후에 zymography를 이용하여 이 추출물의 MMP-2와 -9에 대한 억제효과를 측정한 결과 유의적인 억제효과를 나타내었다. 억제활성 성분을 추적하기 위하여 극성에 따른 추출물의 용매분획을 실시하여 n-hexane, 85% aq. MeOH, n-butanol, 및 water 분획층을 얻었다. 이 4가지 용매분획에 대한 MMP 억제활성을 측정하였으며 측정한 결과 85% aq. MeOH 분획층이 zymography와 RT-PCR 실험에서 MMP-2와 -9에 대해 가장 강한 억제효과를 나타내었다. 이상의 결과는 만형자 추출물이 암전이 억제제 개발을 위한 좋은 원천이 될 수 있는 가능성이 있음을 제시한다.

• Journal of Applied Biological Chemistry
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• 제52권4호
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• pp.180-186
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• 2009

본 연구에서는 염생식물의 한 종인 순비기나무의 추출 분획물 및 화합을 이용하여 항발암성 및 생체방어 물질로서의 유용성을 검토하고자 4종의 인체 암세포(HT1080, AGS, MCF-7 및 HT-29)에 대한 증식억제 효과를 검토하였다. 추출물에 대한 암세포 증식억제능 측정 결과, 10, 50 및 $100\;{\mu}g/mL$의 시료 처리 농도에서 농도 의존적으로 암세포의 생존율이 감소하는 경향을 확인하였으며, 모든 세포주에 대하여 methanol 추출물 보다 methylene chloride 추출물이 뛰어난 효과를 나타냄을 알 수 있었다. 따라서, 물질의 극성도에 따른 암세포 증식억제능을 확인하고자 추출물을 순차적으로 용매 분획하여 $H_2O$, n-BuOH, 85% aq. MeOH 및 n-hexane의 네 가지 분획층을 얻었으며, 추출물과 동일한 조건하에서 암세포 증식억제 효과를 재검토하였다. 그 결과, 비교적 극성이 덜한 85% aq. MeOH 분획층에서 우수한 암세포 증식억제능이 확인되었다. $100\;{\mu}g/mL$의 농도에서 85% aq. MeOH 분획층의 시료를 처리한 결과 HT1080, AGS, MCF-7 및 HT-29 세포는 각각 93.8, 91.6, 90.9% 그리고 82.7%의 강력한 억제 효과를 나타내었으며, 낮은 농도의 시료($10\;{\mu}g/mL$)를 처리한 결과에서도 HT1080과 AGS 세포에서 주목할 만한 억제효과를 보여 주었다. 생리활성 결과를 바탕으로 85% aq. MeOH 분획층의 분리를 통해 1종의 화합물을 분리할 수 있었으며, artemetin으로 알려진 화합물로 확인되었다. 분리된 화합물의 암세포 증식억제 효과를 확인해 본 결과 HT1080 세포에 선택적으로 효과를 보임을 확인하였으며, 그 효과는 항암제인 doxorubicin에 상응하는 결과였다. 따라서 본 연구를 통해 염생식물 순비기나무의 각종 인체 암세포에 대한 높은 증식억제 효과와 함께 암예방을 위한 기능성 소재로서의 개발 가능성이 확인되었다. 추후 순비기나무의 85% aq. MeOH 분획층 및 분리된 화합물에 대한 집중적인 연구를 통해 새로운 생리활성물질의 개발이 기대되어진다.

• KSBB Journal
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• 제27권4호
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• pp.227-231
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• 2012

Whole plants of Zostera asiatica were extracted twice with acetone/methylene chloride (A+M) and methanol (MeOH) in turn. The combined crude extracts were evaporated in vacuo and then the residue was partitioned between water and methylene chloride. The aqueous layer was fractionated into $H_2O$ and n-butanol and then the organic layer was also fractionated into 85% aq. MeOH and n-hexane, successively. The crude extracts and their solvent fractions were evaluated for their inhibitory effect on growth of human cancer cells AGS, HT-29, MCF-7, and HT-1080 cells by MTT reduction assay. Among samples tested, 85% aq. MeOH and n-hexane fractions showed strong cytotoxic effect against AGS, HT-29, and MCF-7 cells. On the other hand, for HT-1080 cell, 85% aq. MeOH fraction exhibited the strongest cytotoxic effect.

• KSBB Journal
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• 제24권2호
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• pp.201-206
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• 2009

본 연구에서는 염생식물의 한 종인 염주괴불주머니의 조추출물 및 용매별 분획물을 이용하여 생리활성물질에 의한 항발암성 물질의 생성 및 생체방어물질로서의 유용성을 검토하고자 5종의 인체암세포 (AGS, HT1080, U-937, MCF-7 및 HT-29)에 대한 증식 억제효과를 검토하였다. 조추출물에 대한 암세포 증식 억제능 측정 결과, 10, 50 및 100 ${\mu}g/ml$의 시료 처리 농도에서 농도 의존적으로 암세포의 생존율이 감소하는 경향을 확인하였으며, 모든 세포주에 대하여 methanol 조추출물 보다 methylene chloride 조추출물이 뛰어난 효과를 나타냄을 알 수 있었다. 따라서, 물질의 극성에 따른 암세포 성장 억제능을 관찰하고자 조추출물을 순차적으로 용매 분획하여 $H_2O$, n-BuOH, 85% aq. MeOH 및 n-hexane의 네 가지 분획층을 얻었으며, 조추출물과 동일한 조건하에서 암세포 증식 억제능을 재검토하였다. 그 결과, n-BuOH 분획층보다 85% aq. MeOH 분획층에서 우수한 암세포 성장 억제능이 확인되어, 이는 비교적 비극성인 methylene chloride 조추출물로부터 활성성분들이 85% aq. MeOH 분획층으로 이동된 것으로 여겨졌다. 특히 100 ${\mu}g/ml$의 85% aq. MeOH 분획층의 시료를 처리한 AGS 인체 위암세포는 93.4%라는 강력한 효력을 기록하였으며, 그 외에도 HT1080 인체 섬유육종 세포가 85.0%, U-937 인체 구성 림프종 세포 71.5%, MCF-7 인체 유방암세포와 HT-29 인체 결장암세포가 각각 51.7% 및 36.8%의 암세포 증식 억제효과를 나타내었다. 이상의 결과로부터 해양생물 중 염생식물의 한 종류인 염주괴불주머니의 높은 항암활성효과와 함께 암예방 기능성 소재로써의 개발 가능성을 확인할 수 있었으며, 특히 염주괴불주머니의 85% aq. MeOH 분획층에 대한 집중적인 연구를 통해 새로운 생리활성물질의 개발이 기대된다.

• 한국식생활문화학회지
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• 제24권6호
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• pp.749-755
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• 2009

We investigated the chemical components of red onion powder dried using the low temperature vacuum method and the inhibitory effects of solvent extracts of the dried red onion powder on the growth of HT-1080 human fibrosarcoma and HT-29 human colon cancer cells and $H_2O_2$-induced oxidative stress. The moisture content of the dried red onion powder was 17.95%, while the vitamin C content was 96 mg/100 g and the total phenols content was 39.1 mg/mL. The inhibitory effects of acetone with methylene chloride (A+M) and methanol (MeOH) extracts of the red onion powder on the growth of HT-1080 and HT-29 cancer cells increased in a dose dependent manner (p<0.05). The inhibitory effect was greater on the growth of HT-29 cells, while the A+M extracts had a higher inhibitory effect than the MeOH extracts. Treatment with the hexane, 85% aq. methanol, butanol and water fractions of the extract led to significant inhibition of the growth of both cancer cell lines (p<0.05). Among the fractions, the hexane and 85% aq. methanol fractions showed a greater inhibitory effect. To determine the protective effect on $H_2O_2$-induced oxidative stress, a DCFH-DA (dichlorodihydrofluorescin diacetate) assay was conducted. All fractions, including the crude extracts of dried red onion, appeared to lead to a significant reduction in the levels of intracellular reactive oxygen species (ROS), and these reductions occurred in a dose dependent fashion (p<0.05). Among the fractions, the 85% methanol fraction showed the greatest protective effect on the production of lipid peroxides.

• 대한한방종양학회지
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• 제4권1호
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• pp.131-146
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• 1998

To examine the effect of Shiquandabutang on the metastasis of cancer, the following experiments were made. Before the main experiments, the cytotoxicity was measured by putting Shiquandabutang sample in HT1080. Then zymography was made to examine the change of gelatinolytic activity. And western blotting was carried out to examine the changes of Fos, Jun, Ets, the transcription factors of MMP-2, MMP-9, and Erk, JNK on signal transduction pathway to AP-1. Third, in vitro invasion assay with transwells coated by collagen and matrigel was carried out. From the results of the above the following conclusions were obtained. 1. The experimental result about cytotoxicity of Shiquandabutang against HT1080 was as below. The stained cell count after being treated by Shiquandabutang sample $400{\mu}g/ml$ for 24 hours was 0.9% of total cells, and the stained cell count by Shiquandabutang sample $100{\mu}g/ml$ was 1.5% of total cells. Both were near the level of control group which showed 0.6% stained. 2. The result of collagenase assay was as below. In Shiquandabutang sample $400{\mu}g/ml$, MMP-2 was reduced as compared with TPA control group, and the band of MMP-9 induced by TPA disappeared. In Shiquandabutang sample $800{\mu}g/ml$, both bands of MMP-2 and MMP-9 disappeared. 3. The results of western blots for Jun, Fos, Ets, Erk, JNK were as below. In Shiquandabutang sample $200{\mu}g/ml$, Ets was reduced, and Fos were increased. 4. The result of invasion assay was as below. The number of cells which migrated across transwell membrane in Shiquandabutang-treated group was less than that of +TPA control group. From the above results, it was concluded that Shiquandabutang might control the appearing and acting of collagenase not by the MMP-2, -9 promoter but by other way.

• 동의생리병리학회지
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• 제23권2호
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• pp.360-367
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• 2009

The purpose of this study was evaluated physiological activity of processed sulfur with Cordyceps militaris mycelium that antioxidative and antiinflammatory effects. Proliferation of processed sulfur (PS) with Cordyceps militaris mycelium was increased in dose-dependent manner. In organic sulfur contents of Cordyceps militalis mycelium fortified processed sulfur, CM+PSH (CM+3000 ppm of PS) was significantly higher than other groups. However, CM+PSL (CM+1500 ppm of PS) was almost changed organic sulfur. Content of total polyphenol compounds was similarity to CM, CM+PSL and CM+PSH. The EDA (electron donating ability) and SOD-like activity was increased in dose-dependent manner and the activity of CM were significantly higher than CM+PSL and CM+PSH. We examined cytotoxicity, nitric oxide production of Raw 264.7 cell and inhibition of HT 1080 cell by MTT assay. CM, CM+PSL and CM+PSH do not have any toxic effects in macrophages (Raw 264.7). And CM+PSL and CM+PSH inhibited the production of nitrite in Raw 264.7 cells activated with LPS. The antitumor effects of processed sulfur with Cordyceps militaris mycelium on HT 1080 cell was indicated a significantly inhibition activity. These results suggested that processed sulfur with Cordyceps militaris mycelium have activities of antioxidant, antiinflammatory effects.

• 동의생리병리학회지
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• 제23권2호
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• pp.473-479
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• 2009

The purpose of this study was evaluated physiological activity effect of organic germanium in culture broth of germanium-fortified Tricholoma matsutake mycelium and germanium-fortified yeast. Proliferation Tricholoma matsutake mycelium and yeast was inhibited by addition of germanium. Contents of organic germanium in Tricholoma matsutake mycelium and yeast was increased in dose-dependent manner. And low concentration(1,000 ppm) of germanium in mycelium was almost changed organic germanium. In the result of antioxidant activity as SOD-like activity, contents of total polyphenol compound and electron donating ability, activity of germanium-fortified Tricholoma matsutake mycelium was higher than that of germanium-fortified yeast. To evaluate of antitumor effects in vitro, we examined nitric oxide production of Raw 264,7 cell and cytotoxicity of HT1080 cell by MTT assay. Nitric oxide production of germanium-fortified Tricholoma matsutake mycelium was shown low level in low concentration(1,000 ppm) than other groups. The anticancer effect of germanium-fortified Tricholoma matsutake mycelium on HT 1080 cell was indicated a strong inhibitory effect in low concentration(1,000 ppm). These results suggest that organic germanium in culture broth of germanium-fortified Tricholoma matsutake mycelium has valuable physiological activities as antioxidant and anticancer effect, and it was higher than that of germanium-fortified yeast.

• 동의생리병리학회지
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• 제24권1호
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• pp.111-117
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• 2010

The purpose of this study was to investigate the effect of garlic(Allium sativum L.) extracts with extraction conditions on antioxidant and anticancer activity. The extracts prepared for garlic by hot temperature extraction (HG), low temperature extraction (LG), UMPM extraction (UG), fermentation (FG) and black garlic hot temperature (BG) method. Content of total polyphenol compound was the BG higher than other extracts. The EDA (electron donating ability) and SOD-like activity was increased in dose-dependent manners, and the activity of BG and UM was significantly higher than LG and FG. We examined cytotoxicity, nitric oxide production of Raw 264.7 cell and inhibition of HT 1080 cell by MTT assay. All extracts does not have any toxic effects in macrophages(Raw 264.7). And UG inhibited the production of nitrite in Raw 264.7 cells activated with LPS. The antitumor effects of LG and UG on HT 1080 cell was indicated a significantly inhibition activity. These results suggested that UG (UMPM extraction of garlic) have activities of antioxidant, anticancer effects.