This study was conducted to evaluate the usability of saffron to improve skin aging and contact dermatitis occurred by permanent oxidative hair dye, ρ-phenilenediamine (PPD). CES was prepared and used at the subsequent experiments.
Contents of total polyphenol and total flavonoid in C.sativus extract were 316.8 μg/mL and 458.5 μg/mL, respectively. DHHP radical scavenging activity and ABTS radical scavenging activity of C.sativus extract were 315.2 μg/mL and 57.1 μg/mL, respectively. Toxicity of CES was not observed at the in vitro test using HaCaT cell. Treatment of CSE showed the recovery of HaCaT cell viability suffered from oxidative stress with dose dependent manner. C.sativus extract-treated samples of 50 μg/mL and of 100 μg/mL recovered to cell viabilities of 80.9% and of 87.9%, respectively, compared with 75.6% of positive control (PC) treated with H₂0₂. Protein expression of Nrf2와 HO-1 decreased by the treatment of H₂0₂ in PC, compared with the expression in negative control (NC). However, treatment of CES increased dose-dependently the expression of Nrf2 and HO-1. Moreover, expression of catalase and SOD, target factors of Nrf2, also increased with the similar tendency of the expression of Nrf2 with dose-independently (p>0.05). In same condition, expression of mRNA of Nrf2, HO-1, catalase and SOD were determined. Expression of mRNA of Nrf2 and HO-1 showed increase with dose dependent manner in the similar tendency at expression of these proteins. Expression of mRNA of catalase and SOD appeared to increase dose-dependently, compared with difference to expression of these proteins. Treatment of PPD caused to decrease of HaCaT cell viability with dose dependent manner. Decreased cell viability was recovered by treatment of CSE dose-dependently. Treatment of PPD caused to increase expression of Bax protein, one of cell death factor, and to decrease expression of Bc12, one of cell viability factor. However, the same treatment of PPD and CSE leaded to decease expression of Bax, and to increase of expression of Bc12. Especially, increase of expression of Bc12 was not dose-dependently, and showed similarly to the value of NC. CSE treated samples of 50 μg/mL and 100 μg/mL decreased statistically at the comparison of the ratio of Bax/Bcl2 values. This result indicated the same tendency of HaCaT cell viability, and that treatment of CSE inhibited cell death occurred by the treatment of PPD and the viability was recovered.
ROS increased to about 2.8 times treated by PPD, and decreased to about 1.4 times added to CSE of 100 μg/mL. This result indicates cell protection effect of C.sativus extract by the inhibition of the generation of ROS by PPD. Expression of Nrf2 and HO-1 in HaCaT cell decreased by treatment of PPD, and was lower than that of NC. By the same treatment with CSE and PPD, difference of the expression of Nrf2 was not observed and that of HO-1 increased with dose dependent manner. These results also appeared at the expression of mRNA of these proteins. Expression of SOD and catalase, representative antioxidant enzymes, increased by treatment of CSE to HaCaT cell suffered from PPD.