Induction of Angiogenesis in Zebrafish Embryos and Proliferation of Endothelial Cells by an Active Fraction Isolated from the Root of Astragalus membranaceus using Bioassay-guided Fractionation
The objective of the study was to identify the active fraction(s) from AR aqueous extract responsible for promoting angiogenesis using bioassay-guided fractionation. The angiogenic activity was screened by monitoring the increase of sprout number in sub-intestinal vessel (SIV) of the transgenic zebr...
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| Format: | Article |
| Language: | English |
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Elsevier
2014-10-01
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| Series: | Journal of Traditional and Complementary Medicine |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2225411016302383 |
| _version_ | 1818200450002845696 |
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| author | Patrick Kwok-Kin Lai Judy Yuet-Wa Chan Hin-Fai Kwok Ling Cheng Hua Yu Ching-Po Lau Ping-Chung Leung Kwok-Pui Fung Clara Bik-San Lau |
| author_facet | Patrick Kwok-Kin Lai Judy Yuet-Wa Chan Hin-Fai Kwok Ling Cheng Hua Yu Ching-Po Lau Ping-Chung Leung Kwok-Pui Fung Clara Bik-San Lau |
| author_sort | Patrick Kwok-Kin Lai |
| collection | DOAJ |
| description | The objective of the study was to identify the active fraction(s) from AR aqueous extract responsible for promoting angiogenesis using bioassay-guided fractionation. The angiogenic activity was screened by monitoring the increase of sprout number in sub-intestinal vessel (SIV) of the transgenic zebrafish embryos after they were treated with 0.06-0.25 mg/ml of AR aqueous extract or its fraction(s) for 96 h. Furthermore, the angiogenic effect was evaluated in treated zebrafish embryos by measuring the gene expression of angiogenic markers (VEGFA, KDR, and Flt-1) using real-time polymerase chain reaction (RT-PCR), and in human microvascular endothelial cell (HMEC-1) by measuring cell proliferation using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 3H-thymidine uptake assay, and cell cycle analysis. A major active fraction (P1-1-1), which was identified as glycoproteins, was found to significantly stimulate sprout formation (2.03±0.27) at 0.125 mg/ml (P<0.001) and up-regulate the gene expression of VEGFA, KDR, and Flt-1 by 2.6-fold to 8.2-fold. Additionally, 0.031-0.125 mg/ml of P1-1-1 was demonstrated to significantly stimulate cell proliferation by increasing cell viability (from 180% to 205%), 3H-thymidine incorporation (from 126% to 133%) during DNA synthesis, and the shift of cell population to S phase of cell cycle. A major AR active fraction consisting of glycoproteins was identified, and shown to promote angiogenesis in zebrafish embryos and proliferation of endothelial cells in vitro. |
| first_indexed | 2024-12-12T02:37:51Z |
| format | Article |
| id | doaj.art-67767f06804e40d48089e4c60b71e7e1 |
| institution | Directory Open Access Journal |
| issn | 2225-4110 |
| language | English |
| last_indexed | 2024-12-12T02:37:51Z |
| publishDate | 2014-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Traditional and Complementary Medicine |
| spelling | doaj.art-67767f06804e40d48089e4c60b71e7e12022-12-22T00:41:14ZengElsevierJournal of Traditional and Complementary Medicine2225-41102014-10-014423924510.4103/2225-4110.139109Induction of Angiogenesis in Zebrafish Embryos and Proliferation of Endothelial Cells by an Active Fraction Isolated from the Root of Astragalus membranaceus using Bioassay-guided FractionationPatrick Kwok-Kin Lai0Judy Yuet-Wa Chan1Hin-Fai Kwok2Ling Cheng3Hua Yu4Ching-Po Lau5Ping-Chung Leung6Kwok-Pui Fung7Clara Bik-San Lau8Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.The objective of the study was to identify the active fraction(s) from AR aqueous extract responsible for promoting angiogenesis using bioassay-guided fractionation. The angiogenic activity was screened by monitoring the increase of sprout number in sub-intestinal vessel (SIV) of the transgenic zebrafish embryos after they were treated with 0.06-0.25 mg/ml of AR aqueous extract or its fraction(s) for 96 h. Furthermore, the angiogenic effect was evaluated in treated zebrafish embryos by measuring the gene expression of angiogenic markers (VEGFA, KDR, and Flt-1) using real-time polymerase chain reaction (RT-PCR), and in human microvascular endothelial cell (HMEC-1) by measuring cell proliferation using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 3H-thymidine uptake assay, and cell cycle analysis. A major active fraction (P1-1-1), which was identified as glycoproteins, was found to significantly stimulate sprout formation (2.03±0.27) at 0.125 mg/ml (P<0.001) and up-regulate the gene expression of VEGFA, KDR, and Flt-1 by 2.6-fold to 8.2-fold. Additionally, 0.031-0.125 mg/ml of P1-1-1 was demonstrated to significantly stimulate cell proliferation by increasing cell viability (from 180% to 205%), 3H-thymidine incorporation (from 126% to 133%) during DNA synthesis, and the shift of cell population to S phase of cell cycle. A major AR active fraction consisting of glycoproteins was identified, and shown to promote angiogenesis in zebrafish embryos and proliferation of endothelial cells in vitro.http://www.sciencedirect.com/science/article/pii/S2225411016302383Astragali RadixGlycoproteinsTraditional Chinese MedicineWound healing |
| spellingShingle | Patrick Kwok-Kin Lai Judy Yuet-Wa Chan Hin-Fai Kwok Ling Cheng Hua Yu Ching-Po Lau Ping-Chung Leung Kwok-Pui Fung Clara Bik-San Lau Induction of Angiogenesis in Zebrafish Embryos and Proliferation of Endothelial Cells by an Active Fraction Isolated from the Root of Astragalus membranaceus using Bioassay-guided Fractionation Journal of Traditional and Complementary Medicine Astragali Radix Glycoproteins Traditional Chinese Medicine Wound healing |
| title | Induction of Angiogenesis in Zebrafish Embryos and Proliferation of Endothelial Cells by an Active Fraction Isolated from the Root of Astragalus membranaceus using Bioassay-guided Fractionation |
| title_full | Induction of Angiogenesis in Zebrafish Embryos and Proliferation of Endothelial Cells by an Active Fraction Isolated from the Root of Astragalus membranaceus using Bioassay-guided Fractionation |
| title_fullStr | Induction of Angiogenesis in Zebrafish Embryos and Proliferation of Endothelial Cells by an Active Fraction Isolated from the Root of Astragalus membranaceus using Bioassay-guided Fractionation |
| title_full_unstemmed | Induction of Angiogenesis in Zebrafish Embryos and Proliferation of Endothelial Cells by an Active Fraction Isolated from the Root of Astragalus membranaceus using Bioassay-guided Fractionation |
| title_short | Induction of Angiogenesis in Zebrafish Embryos and Proliferation of Endothelial Cells by an Active Fraction Isolated from the Root of Astragalus membranaceus using Bioassay-guided Fractionation |
| title_sort | induction of angiogenesis in zebrafish embryos and proliferation of endothelial cells by an active fraction isolated from the root of astragalus membranaceus using bioassay guided fractionation |
| topic | Astragali Radix Glycoproteins Traditional Chinese Medicine Wound healing |
| url | http://www.sciencedirect.com/science/article/pii/S2225411016302383 |
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