Abstract:

【Retraction】

【Abstract】 Objective To evaluate the effect of danshensu on proliferation and apoptosis of M5-stimulated HaCaT cells （a psoriasis-like cell model）, and to explore its relationship with Yes-associated protein （YAP） expression. Methods HaCaT cells were stimulated with M5, a mixture containing 10 μg/L interleukin（IL）-1α, IL-17, IL-22, tumor necrosis factor （TNF）-α and oncostatin M, for 48 hours to establish a psoriasis-like cell model. Then, the cell model was divided into several groups to be treated with 0 （control group）, 0.125, 0.25 and 0.5 mmol/L danshensu respectively, and HaCaT cells receiving no treatment served as the blank control group. Real-time quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of YAP respectively in these groups; methyl thiazolyl tetrazolium （MTT） assay was conducted to estimate the cellular proliferative activity after 24-, 48- and 72-hour treatment with danshensu, flow cytometry to evaluate the effect of danshensu on cell cycle and apoptosis, and Western blot analysis to determine expression of cell cycle-related proteins （cyclin A, cyclin B1, cyclin D, cyclin E） and apoptosis-related proteins （cleaved caspase-3, Bcl-2, BAX, p53 and p21）. One-way analysis of variance was used for comparing means in several groups, and least significant difference （LSD）-t test for multiple comparisons. Results The mRNA and protein expression of YAP significantly differed among the blank control group, control group, 0.125-, 0.25- and 0.5-mmol/L danshensu groups （both P < 0.001）, so did the cellular proliferative activity at 24, 48 and 72 hours （all P < 0.001）. The 0.125-, 0.25- and 0.5-mmol/L danshensu groups all showed significantly decreased mRNA and protein expression of YAP （mRNA: 1.76 ± 0.04, 1.54 ± 0.05, 1.33 ± 0.05 respectively; protein: 1.78 ± 0.06, 1.49 ± 0.32, 1.27 ± 0.04 respectively）, and cellular proliferative activity at 48 hours （1.66 ± 0.04, 1.52 ± 0.02, 1.34 ± 0.04 respectively） compared with the control group （mRNA: 2.04 ± 0.04; protein: 2.10 ± 0.06; cellular proliferative activity: 1.82 ± 0.03; all P < 0.05）. Flow cytometry showed significant differences in the proportions of cells at G0/G1, S and G2/M phases as well as in the apoptosis rates among the above 5 groups （all P < 0.001）. Compared with the control group, the 0.125-, 0.25- and 0.5-mmol/L danshensu groups showed significantly higher proportions of cells at G0/G1 and G2/M phases, but lower proportions of cells at S phase （all P < 0.05）. Additionally, the apoptosis rates were significantly higher in the 0.25- and 0.5-mmol/L danshensu groups than in the control group （both P < 0.05）. Western blot analysis revealed significant differences in the expression of cell cycle-related proteins and apoptosis-related proteins among the above 5 groups （all P < 0.001）. Conclusion Danshensu can inhibit the proliferation of the psoriasis-like cell model and promote its apoptosis, likely by suppressing YAP expression.

Key words: Psoriasis, Danshensu, Keratinocytes, Cell proliferation, Apoptosis, Yes-associated protein