Effect of oleanolic acid on biological function of keloid fibroblasts
10.3760/cma.j.cn114453-20190705-00212
- VernacularTitle:齐墩果酸对瘢痕疙瘩成纤维细胞生物学功能的影响
- Author:
Zhishan XU
1
;
Hongyi WANG
;
Shixiu LIN
;
Jiulong LIANG
;
Quan ZHANG
;
Kai TAO
Author Information
1. 北部战区总医院整形外科，沈阳 110016
- Keywords:
Oleanolic acid;
Keloid;
Fibroblasts;
Cell proliferation;
Apoptosis;
Cell movement
- From:
Chinese Journal of Plastic Surgery
2021;37(4):430-437
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of oleanolic acid on the growth and migration of keloid fibroblasts.Methods:Keloid tissue samples from 9 patients in the Department of Plastic Surgery of General Hospital of Northern Theater were collected and fibroblasts were cultured in vitro. Fibroblasts were treated with different concentrations of oleanolic acid and divided into three groups: control group added 0.9% NaCl; 5 μmol/L oleanolic acid group added 5 μmol/L oleanolic acid; 10 μmol/L oleanolic acid group added 10 μmol/L oleanolic acid. MTT assay was used to detect cell proliferation; flow cytometry was used to detect cell cycle. Annexin V propidium iodide (AV-PI) staining was used to detect cell apoptosis. Transwell assay was used to detect the migration of oleanolic acid. Western blotting and real-time PCR were used to detect the expression of related proteins and mRNA activity. Each group was made in triplicate. Analysis of variance was used to compare the data among the three groups. LSD- t test was used for pairwise comparison, and P<0.05 was considered to be statistically significant. Results:MTT result showed that oleanolic acid could inhibit the proliferation of cells. After 24 hours, the proliferation of cells in 5 μmol/L oleanolic acid group and 10 μmol/L oleanolic acid group were 0.660±0.020 and 0.460±0.020, respectively, compared with 0.780±0.001 in the control group, F=114.4, P<0.001. Compared with the control group, the difference was statistically significant ( t=5.94, P<0.001, t=15.60, P<0.001); flow cytometry showed that the cell cycle G1/S phase transduction was blocked, 5 μmol/L oleanolic acid group and 10 μmol/L oleanolic acid group were significantly inhibited. The percentage of G1 phase cells in the 5 μmol/L oleanolic acid group was significantly higher than that in the control group ( t=3.14, P=0.030, t=6.38, P< 0.001). AⅤ-PI staining showed that the number of apoptotic cells in the 5 μmol/L oleanolic acid group (0.9%) and 10 μmol/L oleanolic acid group (3.4%) was significantly higher than that in the control group (0.4%), and the difference among the three groups was F=119.6, P<0.001. Transwell assay showed that the migration number of cells in 5 μmol/L oleanolic acid group (57.13 ± 2.65) and 10 μmol/L oleanolic acid group (42.15 ± 2.55) was significantly lower than that in control group (72.27± 3.32), F=101.3, P<0.001. Compared with the control group, the difference was statistically significant ( t=6.50, P<0.001, t=14.41, P<0.001). Western blotting showed that oleanolic acid could inhibit the expression of Cyclin D1, Bcl-2, Bax and MMP2. Compared with the control group, 5 μmol/L oleanolic acid t=8.70, P<0.001, t=5.00, P=0.040, t=12.41, P<0.001, t=10.46, P<0.001; compared with the control group, 10 μmol/L oleanolic acid t=31.61, P<0.001, t=23.17, P<0.001, t=12.11, P<0.001, t=44.52, P<0.001. Real-time PCR reaction showed that the mRNA activity levels of Cyclin D1, Bcl-2, Bax, MMP2 were also inhibited. Compared with the control group, 5 μmol/L oleanolic acid t=5.42, P< 0.001, t=3.11, P=0.040, t=16.11, P<0.001, t=11.71, P<0.001; compared with the control group, 10 μmol/L oleanolic acid t=51.78, P<0.001, t=30.89, P<0.001, t=10.64, P<0.001, t=17.10, P< 0.001. Conclusions:Oleanolic acid (5 μmol/L and 10 μmol/L) can inhibit the proliferation and migration of keloid fibroblasts and induce apoptosis of keloid fibroblasts after treating keloid fibroblasts for 24 hours, which can inhibit the growth of keloid and be used for the prevention and treatment of keloid.
