Effect of resveratrol on apoptosis and cell cycle of human keratinocytes irradiated by ultraviolet B
10.35541/cjd.20210059
- VernacularTitle:白藜芦醇对中波紫外线照射的人角质形成细胞凋亡及细胞周期的影响
- Author:
Hongying CHEN
;
Xu CHEN
;
Li LI
;
Heng GU
- From:
Chinese Journal of Dermatology
2021;54(5):408-413
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate the effect of resveratrol on apoptosis and cell cycle of human epidermal keratinocytes (HEKs) after ultraviolet B (UVB) irradiation.Methods:After 12-hour treatment with 0 (control group) , 25, 50, 100, 150 and 200 μmol/L resveratrol, cell counting kit-8 (CCK8) assay, bromodeoxyuridine (BrdU) assay and lactate dehydrogenase (LDH) assay were performed to evaluate the effect of resveratrol on proliferative activity and death of HEKs. Some HEKs were divided into 4 groups: normal control group receiving no treatment, resveratrol group treated with 100 μmol/L resveratrol, UVB group irradiated with 50 mJ/cm 2 UVB, and UVB+resveratrol group irradiated with 50 mJ/cm 2 UVB followed by 12-hour treatment with 100 μmol/L resveratrol. Western blot analysis was conducted to determine the expression of apoptosis-related proteins poly (ADP-ribose) polymerase (PARP) and caspase-3, as well as cell cycle-related proteins cyclin B1 and cyclin E1, and flow cytometry to detect the apoptosis and determine cell cycle distribution. One-way analysis of variance was used for comparisons among multiple groups, and least significant difference- t test for multiple comparisons. Results:The proliferative activity of HEKs significantly differed among the 25-, 50-, 100-, 150-, 200-μmol/L resveratrol groups and control group ( F=46.52, P < 0.001) , and was significantly lower in these resveratrol groups than in the control group (all P < 0.001) ; the DNA synthesis activity of HEKs was significantly lower in the 100-μmol/L resveratrol group (0.761 ± 0.141) than in the control group (2.226 ± 0.141, t=17.92, P < 0.001) ; there was no significant difference in cell death rate among the 25-, 50-, 100- and 200-μmol/L resveratrol groups ( F=1.938, P > 0.05) . After treatment with or without UVB and/or resveratrol, there were significant differences in the apoptosis rate, proportion of cells at G1, G2 and S phases, expression of PARP, caspase-3, cyclin B1 and cyclin E1 among the 4 groups (all P < 0.05) . The UVB group showed significantly increased apoptosis rates (24.69% ± 3.54%) and cleavage levels of PARP and caspase-3 (2.190 ± 0.349, 0.090 ± 0.016, respectively) compared with the normal control group and UVB+resveratrol group (4.00% ± 0.81%, 0.926 ± 0.040, 0.024 ± 0.019, respectively, all P < 0.05) ; the UVB group showed significantly decreased protein expression of cyclin E1 and cyclin B1 (0.116 ± 0.017, 0.775 ± 0.080, respectively) compared with the normal control group, and the UVB+resveratrol group showed significantly increased expression of cyclin E1 (0.287 ± 0.047) , but decreased expression of cyclin B1 (0.504 ± 0.009) compared with the UVB group (all P < 0.05) ; the UVB group showed significantly decreased proportion of S-phase cells, but increased proportion of G1- and G2-phase cells compared with the UVB+resveratrol group (all P < 0.05) . Conclusion:Resveratrol can decrease the proliferative activity of HEKs, inhibit apoptosis induced by UVB radiation, and regulate cell cycle progression.
