신생혈관형성을 억제하는 것은 최근들어 많은 고형암의 치료에 있어서 매우 유용한 접근 방법이다. 현재까지 가장 잘 알려진 접근 방법은 신생혈관형성의 핵심인자인 혈관내피세포성장인자(VEGF)를 표적으로 하는 방법이다. 다양한 화학예방 물질들을 포함하는 많은 자연의 생산물들이나 추출물들은 그들의 항신생혈관형성 성질을 통하여 악성종양의 성장을 억제하고 있다. 황백은 항종양, 항균, 항염증 및 그외 다른 생물학적 작용을 가지고 있는, 오래전부터 널리 사용되어온 한국 전통 약재이다. 우리는 종양의 성장, 침투, 전이에 있어서 매우 중요한 과정인 신생혈관형성에 미치는 황백 온수추출물의 효과를 연구하였다. 황백 온수추출물의 항신생혈관형성 효과를 확인하기 위해서 혈관 내피세포의 성장, 이동, 침투, 관형성 그리고, 자이모그램 분석을 수행하였으며, 흰쥐 대동맥 주변 미세혈관 발아실험을 진행하였다. 그 결과, 황백 온수추출물은 혈관내피세포성장인자(VEGF)에 의해 유도되는 혈관내피세포의 성장, 이동, 침투, 관형성 그리고, 대동맥의 혈관발아를 억제하는 효과를 in vitro와 ex vivo 실험을 통해서 확인하였다. 또한, 황백 추출물은 VEGF에 의해 유도되는 기질금속단백질분해효소 (MMP)-2와 -9의 활성화를 저해하였다. 본 연구 결과들은 황백 온수추출물의 신생혈관형성 억제작용이 암과 같은 혈관신생과 관련된 질병을 치료하는데 좋은 소재가 될 수 있음을 시사한다
신생혈관형성을 억제하는 것은 최근들어 많은 고형암의 치료에 있어서 매우 유용한 접근 방법이다. 현재까지 가장 잘 알려진 접근 방법은 신생혈관형성의 핵심인자인 혈관내피세포성장인자(VEGF)를 표적으로 하는 방법이다. 다양한 화학예방 물질들을 포함하는 많은 자연의 생산물들이나 추출물들은 그들의 항신생혈관형성 성질을 통하여 악성종양의 성장을 억제하고 있다. 황백은 항종양, 항균, 항염증 및 그외 다른 생물학적 작용을 가지고 있는, 오래전부터 널리 사용되어온 한국 전통 약재이다. 우리는 종양의 성장, 침투, 전이에 있어서 매우 중요한 과정인 신생혈관형성에 미치는 황백 온수추출물의 효과를 연구하였다. 황백 온수추출물의 항신생혈관형성 효과를 확인하기 위해서 혈관 내피세포의 성장, 이동, 침투, 관형성 그리고, 자이모그램 분석을 수행하였으며, 흰쥐 대동맥 주변 미세혈관 발아실험을 진행하였다. 그 결과, 황백 온수추출물은 혈관내피세포성장인자(VEGF)에 의해 유도되는 혈관내피세포의 성장, 이동, 침투, 관형성 그리고, 대동맥의 혈관발아를 억제하는 효과를 in vitro와 ex vivo 실험을 통해서 확인하였다. 또한, 황백 추출물은 VEGF에 의해 유도되는 기질금속단백질분해효소 (MMP)-2와 -9의 활성화를 저해하였다. 본 연구 결과들은 황백 온수추출물의 신생혈관형성 억제작용이 암과 같은 혈관신생과 관련된 질병을 치료하는데 좋은 소재가 될 수 있음을 시사한다
Blocking new blood-vessel formation (angiogenesis) is now recognized as a useful approach to the therapeutic treatment of many solid tumors. The best validated approach to date is to target the vascular endothelial growth-factor (VEGF) pathway, a key regulator of angiogenesis. Many natural products ...
Blocking new blood-vessel formation (angiogenesis) is now recognized as a useful approach to the therapeutic treatment of many solid tumors. The best validated approach to date is to target the vascular endothelial growth-factor (VEGF) pathway, a key regulator of angiogenesis. Many natural products and extracts that contain a variety of chemopreventive compounds have been shown to suppress the development of malignancies through their anti-angiogenic properties. Phellodendron amurense, which is widely used in Korean traditional medicine, has been shown to possess antitumor, antimicrobial, and anti-inflammatory properties, among others. The present study investigated the effects of P. amurense hot-water extract (PAHWE) on angiogenesis, a key process in tumor growth, invasion, and metastasis. To investigate PAHWE’s anti-angiogenic properties, this study’s authors performed an analysis of angiogenesis and endothelial-cell proliferation, migration, invasion, and tube formation, as well as zymogram assays and the rat aortic ring-sprouting assay. PAHWE inhibited cell growth, mobility, and vessel formation in response to VEGF in vitro and ex vivo. Furthermore, it reduced VEGF-induced intracellular signaling events, such as the activation of matrix metalloproteinases (MMPs) -2 and -9. These results indicate that PAHWE’s anti-angiogenic properties might lead to the development of potential drugs for treating angiogenesis-associated diseases such as cancer.
Blocking new blood-vessel formation (angiogenesis) is now recognized as a useful approach to the therapeutic treatment of many solid tumors. The best validated approach to date is to target the vascular endothelial growth-factor (VEGF) pathway, a key regulator of angiogenesis. Many natural products and extracts that contain a variety of chemopreventive compounds have been shown to suppress the development of malignancies through their anti-angiogenic properties. Phellodendron amurense, which is widely used in Korean traditional medicine, has been shown to possess antitumor, antimicrobial, and anti-inflammatory properties, among others. The present study investigated the effects of P. amurense hot-water extract (PAHWE) on angiogenesis, a key process in tumor growth, invasion, and metastasis. To investigate PAHWE’s anti-angiogenic properties, this study’s authors performed an analysis of angiogenesis and endothelial-cell proliferation, migration, invasion, and tube formation, as well as zymogram assays and the rat aortic ring-sprouting assay. PAHWE inhibited cell growth, mobility, and vessel formation in response to VEGF in vitro and ex vivo. Furthermore, it reduced VEGF-induced intracellular signaling events, such as the activation of matrix metalloproteinases (MMPs) -2 and -9. These results indicate that PAHWE’s anti-angiogenic properties might lead to the development of potential drugs for treating angiogenesis-associated diseases such as cancer.
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문제 정의
The extract exhibits various biological effects, such as an inhibitory ef- fect on prostatic contractility [49], an anti-osteoarthritic ef- fect [23], an anti-inflammatory property [48], and anticancer activity [16, 18, 26], but antiangiogenic effects have not been reported yet. This present study aimed to elucidate the ef- fect of the hot water extract of P. amurense on angiogenesis. We tested cell biologic functions such as extracellular ma- trix degradation, cell motility, cell proliferation, cell in- vasion and tube formation in vitro, as well as ex vivo neo- vessel formation associated with angiogenesis.
제안 방법
In order to evaluate the antiangiogenic properties of PAHWE, we performed the rat aortic ring assay with 10 and 25 μg/ml PAHWE. The rat aortic ring sprouting assay is a widely used ex vivo anti-angiogenic model that mimics several stages of angiogenesis, including vascular endothelial cell proliferation, migration and tube formation.
4). The final step in this study was to test if the PAHWE can exhibit its antiangiogenic properties ex vivo, as it did in in vitro assays. The VEGF-induced neovessel formation from rat aorta was significantly inhibited by treatment with PAHWE (Fig.
To determine whether PAHWE was capable of influencing endothelial cell migration, we further examined the anti-migration activity of PAHWE on HUVECs. We observed that PAHWE treatment remarkably inhibited VEGF-in-duced endothelial cell migration in comparison with corresponding untreated HUVECs in a non-cytotoxic range of concentrations (Fig.
In the absence of PAHWE, the HUVECs stimulated with VEGF constructed strong and thick vessel tubes by connecting to neighboring cells, but in the presence of PAHWE, in-tercellular connections were limited and the HUVEC cells failed to form tubes. To quantify the degree of tube formation, photographs were taken under an inverted micro- scope and the tube area was calculated using image analysis software. This result suggests that PAHWE may have an inhibitory effect on angiogenesis
an essential pattern in angiogenesis. To verify whether PAHWE could also down-regulate the expression of the pro-angiogenic MMPs, we quantified the extracellular levels of MMP-2 and -9 that were released by HUVECs using a zymogram assay. Both MMP-2 and -9 can hydrolyze gelatin substrates incorporated into an SDS-PAGE gel, and gelatin hydrolysis by MMP-2 and -9 can be visualized by Coomassie staining.
대상 데이터
HUVECs were purchased from American Type Culture Collection (ATCC®, Manassas, VA, USA). HUVECs at early passages (passages 6-10) were used in all experiments and were cultured on gelatin-coated plates in M199 medium (GIBCO®, Grand Island, NY, USA) supplemented with 20% fetal bovine serum (FBS) (GIBCO®, Gaithersburg, MD, USA), 3 ng/ml basic fibroblast growth factor (KOMABIOTECH, Seoul, Korea), 100 unit/ml penicillin, 100 mg/ml strepto- mycin (GIBCO®, Grand Island, NY, USA), and 5 unit/ml heparin (Sigma-Aldrich, St.
The cell suspensions were in- cubated for 30 min at 37℃ in M199 medium (1% FBS) con- taining various concentrations of PAHWE and then seeded into the inserts at 5×104 cells (100 μl)/insert. M199 medium (1% FBS, 600 μl) containing VEGF (20 ng/ml) was added into the chamber as a chemoattractant. The plate was in- cubated for 4 hr at 37℃ in a humidified atmosphere of 5% CO2.
Louis, MO, USA) and incubated at 37℃ in 5% CO2 in air. Seven-week-old Sprague-Dawley rats were purchased from Orient Co. (Sungnam, Korea) and were maintained on standard chow and water ad libitum.
Matrigel basement membrane ma- trix was obtained from BD Biosciences (Bedford, MA, USA). hVEGF165, recombinant human VEGF165 with rat cross re- activity, was purchased from PEPRO TECH (Rocky Hill, NJ, USA). The cell culture reagents and most other biochemical reagents were purchased from Sigma-Aldrich (St.
데이터처리
The data were analyzed for statistically significant differ- ences between groups by using the Student’s t-test and one-way analysis of variance (ANOVA). A p-value lower than 0.
성능/효과
assay. Results showed that the invasive activity of HUVECs in response to VEGF was significantly inhibited in a dose-dependent manner by PAHWE, whereas no suppressive activity was observed in cells treated with PAHWE alone (Fig. 3A).
5). The results show that PAHWE inhibited many cru- cial steps of the angiogenesis process; therefore, this plant may provide a new source of antiangiogenic agents as po- tential candidates for treatment of angiogenesis related dis- eases, such as cancer, psoriasis, rheumatoid arthritis and di- abetic retinopathy.
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