Inhibitory effect of chlorogenic acid on senescence of human skin fibroblasts
10.3760/cma.j.issn.0412-4030.2015.12.005
- VernacularTitle:绿原酸抗人皮肤成纤维细胞衰老的研究
- Author:
Ting CHEN
;
Zhimao JIANG
;
Bo YU
;
Gang MA
- Publication Type:Journal Article
- Keywords:
Skin aging;
Fibroblasts;
Glyoxal;
Chlorogenic acid;
Cell aging;
Cell proliferation;
p16INK4a mRNA
- From:Chinese Journal of Dermatology
2015;(12):849-852
- CountryChina
- Language:Chinese
-
Abstract:
[Abstract ] Objective To evaluate the inhibitory effect of chlorogenic acid on senescence of human skin fibroblasts (HSFs). Methods Fibroblasts isolated from human foreskin were treated with 1 mmol/L glyoxal in vitro to develop a model for cellular senescence. In order to select effective concentrations of chlorogenic acid, some HSFs were treated with 1 mmol/L glyoxal alone or in combination with chlorogenic acid at different concentrations (5, 10, 20, 40, 80 μmol/L)for 3 days, with those receiving no treatment serving as the blank control group. Then, methyl thiazolyl tetrazolium (MTT)assay was performed to evaluate the proliferative activity of HSFs. Some HSFs were divided into 5 groups to be cultured alone(blank control group), or treated with 1 mmol/L glyoxal(glyoxal group)or the combination of 1 mmol/L glyoxal and chlorogenic acid at effective concentrations of 10, 20 and 40 μmol/L (glyoxal + chlorogenic acid groups). Senescence associated β-galactosidase (SA-β-gal)staining and real-time fluorescence-based quantitative PCR were conducted to determine the percentage of senescent cells and expression level of p16INK4a mRNA respectively. Statistical analysis was carried out by one-way analysis of variance followed by the least significant difference(LSD)-t test. Results Compared with the blank control group, the glyoxal group showed significantly decreased cellular proliferative activity of HSFs (55.65% ± 2.00% vs. 100% ± 6.90%, P < 0.01), while chlorogenic acid increased the proliferative activity of HSFs in a dose-dependent manner, and the increase reached a peak at 40 μmol/L. Concretely speaking, the glyoxal + 10-, 20-, 40-, 80-μmol/L chlorogenic acid groups all significantly differed from the glyoxal group in cellular proliferative activity (60.75% ± 1.32%, 67.65% ± 1.90%, 75.71% ± 3.25% and 75.69% ± 2.38% vs. 55.65%± 2.00%, all P < 0.05), but no significant difference was observed between the glyoxal group and glyoxal + 5-μmol/L chlorogenic acid group or between the glyoxal + 40-μmol/L chlorogenic acid group and glyoxal + 80-μmol/L chlorogenic acid group (both P > 0.05). Therefore, 10 - 40 μmol/L was selected as the effective concentrations of chlorogenic acid. The glyoxal group showed significant increases in the percentage of senescent (SA-β-gal-positive)cells (35.65% ± 2.24% vs. 13.00% ± 2.22%, P < 0.01)and expression level of p16INK4a mRNA (2-ΔΔCt: 1.00 ± 0.06 vs. 0.26 ± 0.05, P <0.01)compared with the blank control group, while the glyoxal + 10-, 20-, 40-μmol/L chlorogenic acid groups showed significantly decreased percentage of senescent cells (31.50% ± 2.13% , 22.31% ± 3.11% and 19.32% ± 3.01%respectively)and expression level of p16INK4a mRNA (2-ΔΔCt: 0.88 ± 0.08, 0.73 ± 0.06 and 0.68 ± 0.04 respectively) compared with the glyoxal group (all P < 0.05). Additionally, the percentage of senescent cells decreased with the increase in chlorogenic acid concentrations in the glyoxal + chlorogenic acid groups. Conclusion Chlorogenic acid can protect HSFs from glyoxal-induced senescence.
