Korean J Physiol Pharmacol.  2023 Jan;27(1):105-112. 10.4196/kjpp.2023.27.1.105.

Antioxidation and anti-inflammatory effects of gamma-irradiated silk sericin and fibroin in H ₂O₂-induced HaCaT Cell

• Choi JH
• Lee S
• Han HJ
• Kwon J

• KMID: 2537508
• DOI: http://doi.org/10.4196/kjpp.2023.27.1.105

Abstract

Oxidative stress in skin cells can induce the formation of reactive oxygen species (ROS), which are critical for pathogenic processes such as immunosuppression, inflammation, and skin aging. In this study, we confirmed improvements from gamma-irradiated silk sericin (I-sericin) and gamma-irradiated silk fibroin (I-fibroin) to skin cells damaged by oxidative stress. We found that I-sericin and I-fibroin effectively attenuated oxidative stress-induced ROS generation and decreased oxidative stress-induced inflammatory factors COX-2, iNOS, tumor necrosis factor-α, and interleukin-1β compared to the use of non-irradiated sericin or fibroin. I-sericin and Ifibroin effects were balanced by competition with skin regenerative protein factors reacting to oxidative stress. Taken together, our results indicated that, compared to non-irradiated sericin or fibroin, I-sericin, and I-fibroin had anti-oxidation and antiinflammation activity and protective effects against skin cell damage from oxidative stress. Therefore, gamma-irradiation may be useful in the development of cosmetics to maintain skin health.

Keyword

Gamma-irradiation; Silk fibroin; Silk sericin; Skin regeneration

Figure

• 

  Fig. 1 Effect of sericin and I-sericin or fibroin and I-fibroin on cell viability in H2O2-induced HaCaT cells. Cell viability was measured with MTT assays. (A) HaCaT cells treated with indicated concentrations of sericin and I-sericin (10–100 μg/ml) or fibroin and I-fibroin (10–100 μg/ml) for 24 h. (B) HaCaT cells treated with indicated concentrations of H2O2 (0.1–2 mM) for 24 h. (C) HaCaT cells treated with indicated concentrations of sericin and I-sericin or fibroin and I-fibroin (25–100 μg/ml) and 1 mM H2O2 for 24 h. Data are mean ± SEM (n = 3). Different label means a significant difference between each group by one-way ANOVA. *p < 0.05, compared to control. **p < 0.01, compared to control. ***p < 0.001, compared to control. #p < 0.05, compared to H2O2-only group. ##p < 0.01, compared to H2O2-only group. ###p < 0.001, compared to H2O2-only group. $p < 0.05, compared to the sericin- or fibroin-treated group.

• 

  Fig. 2 Effects of sericin and I-sericin or fibroin and I-fibroin on antioxidant enzymes in H2O2 treated HaCaT cells. (A) ROS production of HaCaT cell with (A1) sericin or I-sericin and (A2) fibroin or I-fibroin pretreatment followed by H2O2; (B) SOD activity of HaCaT cell with (B1) sericin or I-sericin and (B2) fibroin or I-fibroin pretreatment followed by H2O2; (C) GSH content of HaCaT cell with (C1) sericin or I-sericin and (C2) fibroin or I-fibroin pretreatment followed by H2O2. ROS, reactive oxygen species; SOD, superoxide dismutase; GSH, glutathione. Different label means a significant difference between each group by one-way ANOVA. **p < 0.01, compared to control. ***p < 0.001, compared to control. #p < 0.05, ##p < 0.01 compared with H2O2-treated only. $p < 0.05, compared to the sericin or fibroin-treated group.

• 

  Fig. 3 Effects of sericin and I-sericin or fibroin and I-fibroin on inflammatory markers in H2O2-treated HaCaT cells. (A) COX-2 expression of HaCaT cell with (A1) sericin or I-sericin and (A2) fibroin or I-fibroin pretreatment followed by H2O2; (B) iNOS expression of HaCaT cell with (B1) sericin or I-sericin and (B2) fibroin or I-fibroin pretreatment followed by H2O2; (C) IL-1β concentration of of HaCaT cell with (C1) sericin or I-sericin and (C2) fibroin or I-fibroin pretreatment followed by H2O2; (D) TNF-α concentration of HaCaT cell with (D1) sericin or I-sericin and (D2) fibroin or I-fibroin pretreatment followed by H2O2. COX-2, cyclooxygenase-2; iNOS, inducible nitric oxide synthase; IL, interleukin; TNF, tumor necrosis factor. Different label means a significant difference between each group by one-way ANOVA. **p < 0.01, and ***p < 0.005 compared to control. #p < 0.05, ##p < 0.01, and ###p < 0.005 compared with H2O2-treated only. $p < 0.05, compared to the sericin or fibroin-treated group.

• 

  Fig. 4 Effect of of sericin and I-sericin or fibroin and I-fibroin on (A) procollagen type (PCOL1), (B) Elastin, and (C) MMP1 protein expression in in H2O2-treated HaCaT cells. (A) PCOL1 expression of HaCaT cell with (A1) sericin or I-sericin and (A2) fibroin or I-fibroin pretreatment followed by H2O2; (B) Elastin expression of HaCaT cell with (B1) sericin or I-sericin and (B2) fibroin or I-fibroin pretreatment followed by H2O2; (C) MMP1 expression of of HaCaT cell with (C1) sericin or I-sericin and (C2) fibroin or I-fibroin pretreatment followed by H2O2. Different label means a significant difference between each group by one-way ANOVA. ***p < 0.005 compared to control. #p < 0.05, ##p < 0.01, and ###p < 0.005 compared with H2O2-treated only. $p < 0.05, compared to the sericin or fibroin-treated group.

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