Effect of Periplaneta Aamericana Polypeptide on Angiogenesis

Hong-han ZHANG; Rui ZHANG; Fa-lu YUAN; Ting LI; Fang PENG

doi:10.13422/j.cnki.syfjx.20190725

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Effect of Periplaneta Aamericana Polypeptide on Angiogenesis

Pharmacology | 更新时间：2021-02-09

- Effect of Periplaneta Aamericana Polypeptide on Angiogenesis
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 8, Pages: 60-70(2019)
- 作者机构：
  
  1.大理大学　药学与化学学院，云南　大理　 671000；
  2.云南新兴职业学院，昆明　 650200；
  3.云南省药物研究所，昆明　 650111
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20190725
  CLC： R22;R242;R2-031;R285.5;R735.7
- Received：16 December 2018，
  
  Published Online：20 December 2018，
  
  Published：20 April 2019
- 稿件说明：
移动端阅览
Hong-han ZHANG, Rui ZHANG, Fa-lu YUAN, et al. Effect of Periplaneta Aamericana Polypeptide on Angiogenesis[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(8): 60-70.
DOI：

Hong-han ZHANG, Rui ZHANG, Fa-lu YUAN, et al. Effect of Periplaneta Aamericana Polypeptide on Angiogenesis[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(8): 60-70. DOI： 10.13422/j.cnki.syfjx.20190725.

摘要

目的：

探讨美洲大蠊多肽对血管生成的作用，探讨其机制。

方法：

以人脐静脉内皮细胞HUVECs和人肝癌HepG2细胞为研究对象，采用噻唑蓝(MTT)比色法和细胞划痕实验，观察不同质量浓度(6.25

12.5

100 mg·L

-1

)的美洲大蠊多肽(PAP-1～3)，脱脂膏和美洲大蠊多肽前体物CⅡ-3对HUVECs增殖、迁移的影响，另设正常组和沙利度胺组；采用小管形成实验检测各组HUVECs小管形成能力；通过细胞间黏附实验，观察不同质量浓度(25

100 mg·L

-1

)的美洲大蠊提取物处理后人肝癌HepG2细胞同HUVECs之间黏附能力；利用免疫细胞化学染色和酶联免疫吸附测定(ELISA)检测各组HUVECs中血管内皮生长因子(VEGF)蛋白表达。

结果：

MTT比色法结果显示，多肽均能抑制HUVECs的增殖(

0.05)，呈剂量依赖关系。不同质量浓度多肽分别作用24

72 h

PAP-2的抑制作用优于PAP-1

PAP-3(

0.05)。不同质量浓度的C Ⅱ-3和脱脂膏作用

HUVECs的存活率明显升高。细胞划痕实验结果显示多肽均能抑制HUVECs迁移(

0.05)，且随药物浓度的增加，抑制迁移的作用也越强。其中PAP-2的作用优于PAP-1和PAP-3(

0.05)；CⅡ-3在一定程度上能抑制HUVECs迁移，但是剂量越大，抑制作用越弱；脱脂膏作用后

HUVECs迁移能力增强。小管生成实验结果显示，多肽均能抑制HUVECs小管形成(

0.05)，且随着给药浓度的增大，抑制作用越强。其中

PAP-2对小管形成的抑制作用优于PAP-1和PAP-3(

0.05)。CⅡ-3和脱脂膏一定程度上促进HUVECs小管的形成。细胞间黏附实验结果显示，多肽均能阻断HepG2同HUVECs之间的黏附(

0.05)，其中PAP-2对HepG2同HUVECs间黏附的阻断作用强于PAP-1和PAP-3(

0.05)。相反

CⅡ-3和脱脂膏在一定程度上却能促进HepG2同HUVECs间黏附。免疫细胞化学染色和ELISA实验结果显示，多肽均能够下调HUVECs细胞中VEGF含量(

0.05)，且呈浓度依赖性。其中

PAP-2对VEGF的下调作用优于PAP-1和PAP-3(

0.05)。而CⅡ-3和脱脂膏能上调HUVECs中VEGF的表达。

结论：

美洲大蠊多肽对HUVECs的侵袭和转移，分化形成小管的能力有一定抑制作用，并能下调细胞中VEGF蛋白的表达。其中，美洲大蠊多肽的作用优于CⅡ-3和脱脂膏，多肽中PAP-2的作用较优。

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

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

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 (

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 (

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 (

0.05)

and the effect of PAP-2 was better than that of PAP-1 and PAP-3 (

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 (

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 (

0.05)

and the effect of PAP-2 was better than that of PAP-1 and PAP-3 (

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 (

0.05)

in which effect of PAP-2 was better than that of PAP-1 and PAP-3 (

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.

关键词

Keywords

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