JOURNAL OF COSMETIC SCIENCE 24 through sacran or HA-treated membrane fi lters had signifi cantly reduced CYP1A1 mRNA expression levels (Figure 4). On the other hand, when comparing sacran and HA, sacran showed a more signifi cant downregulation of CYP1A1 mRNA expression levels than HA (Figure 4). AMELIORATION OF ADVERSE EFFECTS INDUCED BY TOBACCO SMOKE ON HaCaT KERATINOCYTES Treatment of HaCaT keratinocytes with tobacco smoke showed high cytotoxicity, but sacran or HA signifi cantly reduced the cytotoxicity of tobacco smoke. Compared with the Figure 3. Trapping effects of polysaccharide-treated fi lters on ACs and BaP in tobacco smoke. ACs and BaP in PBS(-) diffused with tobacco smoke passed through fi lters treated with or without polysaccharides were quantifi ed as follows: ACs were estimated by FI (Ex 470 nm, Em 550 nm) with NBD-hydrazine in the pres- ence of trifl uoroacetic acid. BaP was quantifi ed by FI (Ex 360 nm, Em 450 nm). Calibration curves were prepared, and their concentrations were quantifi ed. Each value represents the mean ± SD of three experi- ments. Wilcoxon rank-sum test, ***p 0.001. Trapping effi ciency is expressed as a percentage versus the value of the H2O-treated membrane fi lter diffused with tobacco smoke (Control).

SACRAN PROTECTS SKIN AGAINST POLLUTANTS 25 Figure 4. Suppressive effects of polysaccharides on CYP1A1 mRNA expression levels induced by tobacco. HaCaT keratinocytes were cultured in DMEM containing PBS diffused with tobacco smoke through fi lters treated with or without sacran or HA. After 24 h, total RNA was extracted and CYP1A1 mRNA levels were determined by real-time PCR. Each value represents the mean ± SD of four experiments. **p 0.01. Control(-) denotes sham-treated cells (PBS(-) nontreated with tobacco smoke) and Control(+) denotes cells treated with tobacco smoke diffused through a nontreated fi lter.