Effect of hemoporfin-photodynamics on VEGF-A/MAPK/ERK pathway in vascular endothelial cells
10.3760/cma.j.cn114453-20200304-00104
- VernacularTitle:海姆泊芬-光动力对血管内皮细胞VEGF-Α/MAPK/ERK通路的作用
- Author:
Qichao LIU
1
;
Yingjuan MEI
;
Youquan LUO
;
Mengxue ZHENG
;
Rui YIN
Author Information
1. 陆军军医大学西南医院皮肤科，重庆 400038
- Keywords:
Hemoporfin;
Photodynamic therapy;
MAPK pathway;
Port wine stain;
Human umbilical vein endothelial cells
- From:
Chinese Journal of Plastic Surgery
2020;36(4):385-391
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the molecular mechanism of Hemoporfin-mediated photodynamic therapy on vascular endothelial cells.Methods:Human umbilical vein endothelial cells (HUVEC) were cultured in vitro and divided into four groups: PDT group(Hemoporfin concentration: 5 μg/ml, light fluences: 4 J/cm 2), drug group (only Hemporfin: 5 μg/ml), light group(only irradiation by 4J/cm 2 light), and blank control (no drug, no light). The cell viability and proliferation were detected by cck-8 cytotoxicity test and Brdu testafter different treatments as mentioned above. Expression levels of VEGF-A/MAPK/ERK pathway related molecules in the cells before and after photodynamic treatment were detected by real-time quantitative PCR, Western blot and immunofluorescence staining. Results:Compared with the black control group, the cell viability[(0.45±0.08)vs(1.02±0.11), t=12.02, P<0.05] and cell proliferation level [(0.42±0.02)vs(1.00±0.01), t=31.20, P<0.05]were significantly decreased in PDT group.The mRNA expression levels, including Ras[(0.62±0.02)vs(1.05±0.03), t=10.35, P<0.05], c-Raf [(0.72±0.04)vs(1.00±0.05), t=7.35, P<0.05], Mek[(0.73±0.12)vs(1.15±0.04), t=7.74, P<0.05], Erk [(0.56±0.11)vs(1.02±0.03), t=5.56, P<0.05], VEGF-Α [(0.34±0.04)vs(1.02±0.07), t=7.59, P<0.05], and VEGFR2[(0.54±0.05)vs(1.00±0.03), t=5.34, P<0.05] were significantly decreased. The proteinphosphorylation level of c-Raf[(0.44±0.02)vs(1.02±0.05), t=46.7, P<0.05], Mek[(0.72±0.05)vs(1.05±0.04), t=5.35, P<0.05], Erk[(0.62±0.15)vs(1.03±0.03), t=8.58, P<0.05] and the proteinexpression level of VEGF-A[(0.64±0.03)vs(1.03±0.04), t=21.65, P<0.05] were significantly down-regulated in PDT group compared with the black control group. Compared with the blank control group, there were no significant differences expression between the drug group and the light group at cell activity, molecular proliferation level and molecular expressions. Conclusions:HMME-PDT inhibits the activity and proliferation of vascular endothelial cells by inhibiting the expression of the VEGF-A/MAPK/ERK pathway to achieve the purpose of inhibiting vascular hyperplasia and repair.
