Effect of Periplaneta Aamericana Polypeptide on Angiogenesis
10.13422/j.cnki.syfjx.20190725
- VernacularTitle: 美洲大蠊多肽对血管生成的影响
- Author:
Hong-han ZHANG
1
;
Rui ZHANG
1
;
Fa-lu YUAN
2
;
Ting LI
3
;
Fang PENG
1
Author Information
1. College of Pharmacy and Chemistry, Dali University, Dali 671000, China
2. Yunnan Xinxing Occupations Institute, Kunming 650200, China
3. Yunnan Institute of Materia Medica, Kunming 650111, China
- Publication Type:Research Article
- Keywords:
Periplaneta Americana polypeptide;
human umbilical vein endothelial cells(HUVECs);
HepG2 cells;
proliferation;
migration;
vascular endothelial growth factor (VEGF)
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(8):60-70
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of the Periplaneta Americana polypeptide on the angiogenesis. Method:Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and cell scratch assay were used to observe effect of different concentration (6.25,12.5,25,50,100 mg·L-1) of the Periplaneta Americana polypeptide, CⅡ-3 and skimmed cream on the proliferation and migration of human umbilical vein endothelial cells (HUVECs), and a normal group and a thalidomide group were also established in this study. The tubule formation assay was used to detect the effect of different concentration (25, 50, 100 mg·L-1) of the Periplaneta Americana extracts on the formation of tubules in HUVECs cells. The adhesion between HepG2 cells and HUVECs cells was observed by cell adhesion assay. The expression of vascular endothelial growth factor (VEGF) proteins in HUVECs was detected by immunocytochemical staining and enzyme linked immunosorbent assay (ELISA). Result:MTT results showed that the Periplaneta Americana polypeptide could inhibit the proliferation of HUVECs in a dose-dependent manner (P<0.05). The effect of different concentrations of PAP-2 was better than that of PAP-1 and PAP-3 at 24, 48 72 h (P<0.05). However, the survival rate of HUVECs was significantly increased after treatment with different concentrations of CⅡ-3 and skimmed cream in a time-dose dependent manner. Cell wound scratch assay indicated that migration of HUVECs could be inhibited by Periplaneta Americana polypeptide in a dose-dependent manner (P<0.05), and the effect of PAP-2 was better than that of PAP-1 and PAP-3 (P<0.05). CⅡ-3 could inhibit migration HUVECs to some extent, but the higher the dose was, the weaker the inhibition effect was. Skimmed cream promoted migration of HUVECs in a dose-dependent manner. Tube formation assay revealed that Periplaneta Americana polypeptide could inhibit tube formation of HUVECs, and the inhibitory effect of PAP-2 was better than that of PAP-1 and PAP-3 (P<0.05). CⅡ-3 and skimmed cream promoted the tube formation of HUVECs to a certain extent. Intercellular adhesion assay stated clearly that Periplaneta Americana polypeptide could block the adhesion between HepG2 and HUVECs (P<0.05), and the effect of PAP-2 was better than that of PAP-1 and PAP-3 (P<0.05). However, CⅡ-3 and skimmed cream could promote the adhesion between HepG2 and HUVECs. Results of ELISA assay and immunocytochemical staining indicated that Periplaneta Americana polypeptide could down-regulate the expression of VEGF of HUVECs in a dose-dependent manner (P<0.05), in which effect of PAP-2 was better than that of PAP-1 and PAP-3 (P<0.05). However, CⅡ-3 and skimmed cream could up-regulate the expression of VEGF in HUVECs. Conclusion:The Periplaneta Americana polypeptide can inhibit the invasion, metastasis and tube formation of HUVECs, and down-regulate the expression of VEGF in HUVECs. The effect of Periplaneta Americana polypeptide is better than CⅡ-3 and skimmed cream, and the among the polypeptide, the effect of PAP-2 is superior to the other two.