In vitro와 ex vivo 혈관신생 모델에서 황련 냉수추출물의 신생혈관 억제효과 Antiangiogenic Activity of Coptis chinensis Franch. Water Extract in in vitro and ex vivo Angiogenesis Models원문보기
혈관신생, 즉 새로운 혈관형성은 종양의 성장과 전이에 중요한 역할을 한다고 알려져 있으며, 암 치료에 중요한 목표물이 되고 있다. 이 연구의 목적은 황련 냉수추출물의 혈관신생 억제작용의 효과를 밝히고 항암제로서의 가능성을 평가하고자 한다. Ex vivo rat aortic ring assay 실험결과 신생혈관성장을 억제하는 결과를 확인하였고, 이것을 통해 황련 냉수추출물이 혈관신생과정의 주요한 단계와 내피세포의 증식, 이동, 침투, 혈관내피세포자극인자에 의한 반응으로 모세관 모양의 관 형성작용을 억제함을 알아내었다. 또한 황련 냉수추출물을 처리하였을 때, 세포주기가 억제되고 VEGF에 의한 반응으로 인해 G0/G1 주기에서 S 주기로 가는 과정을 예방하고, VEGF에 의해 활성화가 유도되는 MMP-2, MMP-9이 감소되었다. 따라서 이들의 결과는 황련 냉수추출물이 종양의 발달 단계 중 혈관신생을 방해하는 잠재적인 항암약물의 소재로 고려될 수 있음을 제안한다.
혈관신생, 즉 새로운 혈관형성은 종양의 성장과 전이에 중요한 역할을 한다고 알려져 있으며, 암 치료에 중요한 목표물이 되고 있다. 이 연구의 목적은 황련 냉수추출물의 혈관신생 억제작용의 효과를 밝히고 항암제로서의 가능성을 평가하고자 한다. Ex vivo rat aortic ring assay 실험결과 신생혈관성장을 억제하는 결과를 확인하였고, 이것을 통해 황련 냉수추출물이 혈관신생과정의 주요한 단계와 내피세포의 증식, 이동, 침투, 혈관내피세포자극인자에 의한 반응으로 모세관 모양의 관 형성작용을 억제함을 알아내었다. 또한 황련 냉수추출물을 처리하였을 때, 세포주기가 억제되고 VEGF에 의한 반응으로 인해 G0/G1 주기에서 S 주기로 가는 과정을 예방하고, VEGF에 의해 활성화가 유도되는 MMP-2, MMP-9이 감소되었다. 따라서 이들의 결과는 황련 냉수추출물이 종양의 발달 단계 중 혈관신생을 방해하는 잠재적인 항암약물의 소재로 고려될 수 있음을 제안한다.
Angiogenesis, the formation of new blood vessels, plays an important role in tumor growth and metastasis; therefore, it has become an important target in cancer therapy. Novel anticancer pharmaceutical products that have relatively few side effects or are non-cytotoxic must be developed, and such pr...
Angiogenesis, the formation of new blood vessels, plays an important role in tumor growth and metastasis; therefore, it has become an important target in cancer therapy. Novel anticancer pharmaceutical products that have relatively few side effects or are non-cytotoxic must be developed, and such products may be obtained from traditional herbal medicines. Coptis chinensis Franch. is an herb used in traditional medicine for the treatment of inflammatory diseases and diabetes. However, potential antiangiogenic effects of C. chinensis water extract (CCFWE) have not yet been studied. The purpose of this study was to determine the antiangiogenic effect of CCFWE in order to evaluate its potential for an anticancer drug. We found that the treatment with CCFWE inhibited the major steps of the angiogenesis process, such as the endothelial cell proliferation, migration, invasion, and capillary-like tube formation in response to vascular endothelial growth factor (VEGF), and also resulted in the growth inhibition of new blood vessels in an ex vivo rat aortic ring assay. We also observed that CCFWE treatment arrested the cell cycle at the G0/G1 phase, preventing the G0/G1 to S phase cell cycle progression in response to VEGF. In addition, the treatment reduced the VEGF-induced activation of matrix metalloproteinases 2 and 9. Taken together, these findings indicate that CCFWE should be considered a potential anticancer therapy against pathological conditions where angiogenesis is stimulated during tumor development.
Angiogenesis, the formation of new blood vessels, plays an important role in tumor growth and metastasis; therefore, it has become an important target in cancer therapy. Novel anticancer pharmaceutical products that have relatively few side effects or are non-cytotoxic must be developed, and such products may be obtained from traditional herbal medicines. Coptis chinensis Franch. is an herb used in traditional medicine for the treatment of inflammatory diseases and diabetes. However, potential antiangiogenic effects of C. chinensis water extract (CCFWE) have not yet been studied. The purpose of this study was to determine the antiangiogenic effect of CCFWE in order to evaluate its potential for an anticancer drug. We found that the treatment with CCFWE inhibited the major steps of the angiogenesis process, such as the endothelial cell proliferation, migration, invasion, and capillary-like tube formation in response to vascular endothelial growth factor (VEGF), and also resulted in the growth inhibition of new blood vessels in an ex vivo rat aortic ring assay. We also observed that CCFWE treatment arrested the cell cycle at the G0/G1 phase, preventing the G0/G1 to S phase cell cycle progression in response to VEGF. In addition, the treatment reduced the VEGF-induced activation of matrix metalloproteinases 2 and 9. Taken together, these findings indicate that CCFWE should be considered a potential anticancer therapy against pathological conditions where angiogenesis is stimulated during tumor development.
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문제 정의
This complex multistep process is associated with basal membrane injury, endothelial cell proliferation, migration, invasion, and capillary-like structure formation, ending up in the recruitment and adhesion of pericytes or smooth muscle cells [18]. The aim of the current research was to examine the antiangiogenic effects of CCFWE on the VEGF-induced angiogenic process both in vitro and ex vivo.
가설 설정
Fig. 6. CCFWE inhibits vessel sprouting from rat aorta. Rat aortic rings were placed in Matrigel and treated with 10 or 25 μg/ml CCFWE before the exposure to VEGF (20 ng/ml).
제안 방법
1A. After having established that CCFWE does not affect cell viability, all experiments were performed at non-toxic concentrations of CCFWE. Since angiogenesis begins with local proliferation of endothelial cells in response to a proangiogenic stimulus from VEGF, we first determined the antiproliferative effect of CCFWE on human endothelial cells in vitro.
We then examined the effect of CCFWE on cell cycle progression using fluorescence-activated cell sorting (FACS) analysis. After the treatment of HUVECs with 20 ng/ml VEGF, with or without CCFWE, for 20 hr, the percentage of cells in the G0/G1, S, and G2/M phases was monitored. VEGF induced HUVECs to enter into the S phase, whereas the treatment with CCFWE significantly increased the proportion of cells at G0/G1 (Fig.
Experiments were repeated three times, and the results are shown as the mean ± SD of triplicate determinations.
5). Finally, to verify the antiangiogenic effect of CCFWE, we carried out an ex vivo rat aortic ring sprouting assay. The results showed that the VEGF-stimulated outgrowth and dendritic branching of new blood vessels were potently and dose-dependently inhibited in the rat aortic rings treated with CCFWE (Fig.
A vital step in cell migration and invasion includes the degradation of the extracellular matrix (ECM), and MMPs play a central role in the process [41]. Therefore, we further performed a zymogram assay to examine the effect of CCFWE on the VEGF-stimulated secretion of MMP-2 and MMP-9 into the supernatant of cultured media. Gelatin zymography of the serum-free cultured medium supernatants revealed that the activities of MMP-2 and MMP-9 in VEGFtreated cells were substantially higher than those in untreated cells.
이론/모형
Cell viability was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; USB Corporation, Cleveland, OH, USA) assay. One day before the extract treatment, HUVECs (1×105 cells) were seeded into each well of a gelatin (Sigma)-coated 24-well tissue culture plate.
성능/효과
In conclusion, we demonstrated that a water extract of C. chinensis could be an interesting antiangiogenic candidate that targets the VEGF signaling pathway and may be a potential novel therapeutic agent for the treatment of cancer and other angiogenesis-related diseases.
Finally, to verify the antiangiogenic effect of CCFWE, we carried out an ex vivo rat aortic ring sprouting assay. The results showed that the VEGF-stimulated outgrowth and dendritic branching of new blood vessels were potently and dose-dependently inhibited in the rat aortic rings treated with CCFWE (Fig. 6). Collectively, the inhibition of microvessel outgrowth from rat aortic rings, as well as the inhibition of HUVEC proliferation, migration, invasion, and tube formation on a Matrigel matrix, provides strong evidence of the antiangiogenic effect of CCFWE.
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