Angiotensin (1－7) activates autophagy to regulates apoptosis and angiogenesis of oral submucosal fibroblasts
10.19405/j.cnki.issn1000-1492.2023.03.018
- Author:
Lehong Qiu
1
;
Wei Deng
1
;
Chengwen Gan
1
;
Ying Sun
1
Author Information
1. Dept of Stomatology，Hainan General Hospital， Hainan Hospital Affiliated to Hainan Medical College，Haikou 570311
- Publication Type:Journal Article
- Keywords:
soral submucosal fibrosis;
fibroblasts;
angiotensin ( 1-7);
apoptosis;
autophagy;
angiogenesis
- From:
Acta Universitatis Medicinalis Anhui
2023;58(3):457-464
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the effect of angiotensin ( 1-7) ［Ang( 1-7) ］on the apoptosis and angiogenesis of fibroblasts in the oral submucosal fibrosis ( OSF) ，and to explore the effect preliminarily mechanism．
Methods:Fibroblasts were isolated and cultured from human buccal mucosal tissue，the cell morphology was observed by inverted microscope，and the expression of vimentin was detected by immunofluorescence staining ; areca nut extract (ANE) was used to induce human fibroblasts to simulate the in vitro model of fibroblasts in OSF，the experimental groups included control group ( normally cultured cells) ，ANE group ( 100 μg/ ml ANE cultured cells for 48 hours) ，ANE + low-dose Ang( 1-7) group ( 100 μg/ ml ANE + 10－7 mol /L Ang ( 1-7) cultured cells for 48 h) ， ANE + high-dose Ang( 1-7) group ( 100 μg/ ml ANE + 10－5 mol /L Ang( 1-7) cultured cells for 48 h) ，immunofluorescence staining detected the expression of α-smooth muscle actin ( α-SMA) ，ELISA method detected the content of Collagen I and Collagen Ⅲ in the cell culture supernatant，MTT method detected cell proliferation activity， flow cytometry detected cell apoptosis ，the tubule formation experiment detected the vascularization of human umbilical vein endothelial cell(HUVEC) ; After the mRFP-GFP-LC3 virus was transferred to the cells，the level of autophagy was detected by immunofluorescence staining，Western blot detected the expression of autophagy-related protein Beclin-1 and the ratio of LC3-Ⅱ/ LC3-Ⅰ .
Results:The isolated and cultured cells were in a long spindle shape，and Vimentin was positively expressed，indicating that fibroblasts were successfully isolated ; Compared with the ANE group，the fluorescence expression of α-SMA protein in ANE low dose Ang( 1-7) group and ANE + high dose Ang( 1-7) group significantly decreased，the contents of Collagen I and Collagen Ⅲ in the culture supernatant were reduced (P＜0．05) ，cell proliferation activity decreased (P＜0．05) ，and cell apoptosis rate increased (P < 0．05) ，the cell culture supernatants of the two groups inhibited the angiogenesis of HUVEC (P＜0．05) ，endophagosomes were reduced (P＜0．05) ，Beclin-1 protein expression was reduced (P ＜0．05) ，and the ratio of LC3- Ⅱ / LC3-Ⅰ was down-regulated (P ＜0．05 ) ; in addition ，the effect of ANE + high-dose Ang ( 1-7 ) group was stronger than that of ANE + low-dose Ang( 1-7) group (P＜0．05) ．
Conclusion: Ang( 1-7) can inhibit the activation of fibroblasts induced by ANE，promote cell apoptosis，and reduce the angiogenesis of HUVEC，this mechanism may be related to the regulation of cell autophagy.
- Full text:2024071621001392327血管紧张素(1-7)激活自...变细胞凋亡与血管生成的研究_邱乐宏.pdf
