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. 2018 Aug;18(2):1439-1446.
doi: 10.3892/mmr.2018.9150. Epub 2018 Jun 11.

Protective Effect of Crocin on Ultraviolet B‑induced Dermal Fibroblast Photoaging

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Free PMC article

Protective Effect of Crocin on Ultraviolet B‑induced Dermal Fibroblast Photoaging

Mingwu Deng et al. Mol Med Rep. .
Free PMC article

Abstract

Ultraviolet B (UVB) radiation induces the production of reactive oxygen species (ROS), resulting in the aging of dermal fibroblasts. Crocin, a bioactive constituent of Crocus sativus, possesses anti‑oxidation effects. The purpose of the present study was to evaluate the protective effect of crocin on UVB‑induced dermal fibroblast photoaging. Human dermal fibroblasts were isolated and cultured with different concentrations of crocin prior to and following exposure to UVB irradiation. The senescent phenotypes of cells were evaluated, including cell proliferation, cell cycle, senescence‑associated β‑galactosidase (SA‑β‑gal) expression, intracellular ROS, expression of antioxidant protein glutathione peroxidase 1 (GPX‑1) and extracellular matrix protein collagen type 1 (Col‑1). Crocin rescued the cell proliferation inhibited by UVB irradiation, prevented cell cycle arrest and markedly decreased the number of SA‑β‑gal‑positive cells. In addition, crocin reduced UVB‑induced ROS by increasing GPX‑1 expression and other direct neutralization effects. Furthermore, crocin promoted the expression of the extracellular matrix protein Col‑1. Crocin could effectively prevent UVB‑induced cell damage via the reduction of intracellular ROS; thus, it could potentially be used in the prevention of skin photoaging.

Figures

Figure 1.
Figure 1.
Effects of crocin (0, 12.5, 50 and 100 µm) on the proliferation of dermal fibroblasts.
Figure 2.
Figure 2.
Crocin rescues the cell proliferation inhibited by UVB irradiation. (A) UVB irradiation inhibited fibroblast proliferation in a dose-dependent manner. *P<0.05 vs. control. (B) Crocin rescued the cell proliferation inhibited by UVB irradiation. *P<0.05, as indicated. (C) Morphology of fibroblasts at 72 h post-UVB irradiation. Scale bars, 150 µm. UVB, ultraviolet B; Vit C, vitamin C.
Figure 3.
Figure 3.
Crocin rescues UVB-induced cell cycle arrest. (A) Representative histograms of the cell cycle analysis, as determined by flow cytometry. (B) Statistical analysis of cell cycle distribution from three independent experiments. #P<0.05 vs. control; *P<0.05 vs. UVB. UVB, ultraviolet B; Vit C, vitamin C.
Figure 4.
Figure 4.
Crocin prevents UVB-induced cell aging. (A) Positive SA-β-gal staining indicated aging cells. Scale bars, 100 µm. (B) Percentage of SA-β-gal-positive cells was determined by counting 200 cells per dish. *P<0.05, **P<0.01 and ***P<0.001, as indicated. UVB, ultraviolet B; Vit C, vitamin C; SA-β-gal, β-galactosidase.
Figure 5.
Figure 5.
Crocin reduces UVB-induced intracellular ROS. (A) Intracellular ROS measured by DCFH2-DA staining at 20 min post-UVB irradiation. Scale bars, 500 µm. (B) Intracellular ROS measured by DCFH2-DA staining and flow cytometry. (C) Antioxidant effect of crocin measured by 1,1-diphenyl-2-picrylhydrazyl radical-scavenging activity. (D) Western blot analysis of (E) GPX-1 expression at 72 h post-UVB irradiation. *P<0.05, **P<0.01 and ***P<0.001, as indicated. UVB, ultraviolet B; Vit C, vitamin C; ROS, reactive oxygen species; GPX-1, glutathione peroxidase 1; DCFH2-DA, 2′,7′-dichlorodihydrofluoresce in diacetate.
Figure 6.
Figure 6.
Crocin promotes the expression of collagen type 1. (A) Reverse transcription-quantitative polymerase chain reaction analysis of pro-collagen I at 72 h post-UVB irradiation. (B) Western blotting analysis of (C) collagen type 1 at 72 h post-UVB irradiation. *P<0.05, **P<0.01 and ***P<0.001, as indicated. UVB, ultraviolet B; Vit C, vitamin C; col 1, collagen type 1.

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