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The Activity and Mechanism Research of Total Saponins from Albizia julibrissin and Vaccaria segetalis on the Anti-angiogenesis

Author: MaLiPing
Tutor: QiuLiYing
School: Jiangnan University
Course: Microbial and Biochemical Pharmacy
Keywords: Albizia julibrissin Durazz Vaccaria segetalis (Neck.)Garcke total saponins endothelial cells activity mechanism
CLC: R285
Type: Master's thesis
Year: 2013
Downloads: 0
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Abstract


The tumor growth and metastasis rely on oxygen and nutrition supplied by the vessels,so the angiogenesis plays an important role in the process of tumor progression, including thedegradation of extracellular matrix, migration, proliferation of endothelial cells, tubeformation and maturation of vessels. Therefore, inhibiting any part of angiogenic processcould restrict the tumor growth. We used HMEC-1model in vitro and Matrigel plug andtumor model in vivo to detect the effects and mechanism of total saponins of Albiziajulibrissin Durazz and Vaccaria segetalis (Neck.) Garcke on the anti-angiogenesis.bFGF could facilitate the HMEC-1cells growth. AJBE and VSET inhibitedbFGF-induced proliferation of HMEC-1cells with IC50value of5.89±0.33μg/mL and4.67±0.25μg/mL, respectively. AJBE and VSET also suppressed the migration and tubeformation of HMEC-1cell for48h. Trypan blue stain was used to detect the cell viability,which showed there was no necrosis of HMEC-1with treatment of AJBE(5μg/mL). However,necrosis was detected with treatment of VSET(4μg/mL).Then the flow cytometry was applied to investigate HMEC-1cell cycle and apoptosis inAJBE and VSET-treated group, and Annexin V/PI double staining was further to testapoptotic variation. AJBE and VSET could make the S phase arrested and induced apoptosisin HMEC-1cells in concentration-dependent-manner. Meanwhile,VSET(4μg/mL) inducedthe necrosis of HMEC-1. West blotting was used to analyze signal pathway of PI3K/AKT,MAPKs/ERKs and p38MAPK. AJBE and VSET could restrict phosphorylation of bFGFR,and reduced the target protein of pAKT, pERK and pP38expression after48h. Caspase-3protein, which was related to the apoptosis, was activated by AJEB and VSET.In vivo, AJBE and VSET inhibited bFGF-induced angiogenesis significantly in Matrigelplug model by daily oral administration after14d. Immunohistochemisty (IHC) methodindicated that AJBE and VSET disrupted the expression of pbFGFR, pAKT and pERK. H22tumor model was established to study the effects of medical on the tumor angiogenesis. Theinhibition rate of tumor growth with the treatment of AJBE and VSET at low and high dosewas32.74%,37.37%,10.96%and26.33%, respectively. MVD was inhibited by AJBE indose–dependent-manner, but VSET restricted MVD only at high dose. HE staining provedthat there were pyknosis, necrosis and immersion of inflammatory cells in tumor tissue athigh concentration of AJBE and VSET. IHC method certificated that AJBE inhibited theangiogenesis of tumor through PI3K/AKT and MAPKs/ERKs pathway, while VSET exertedthe anti-angiogenesis through PI3K/AKT, MAPKs/ERKs and p38MAPK pathway. VSETactivated p38MAPK signal pathway that was maybe related with haemorrhagic necrosis ofintestine.

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