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Effect of danshensu on Yes-associated protein expression in as well as on proliferation and apoptosis of a psoriasis-like cell model / 中华皮肤科杂志
Chinese Journal of Dermatology ; (12): 452-458, 2020.
Article in Chinese | WPRIM | ID: wpr-870296
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.
Full text: Available Index: WPRIM (Western Pacific) Type of study: Prognostic study Language: Chinese Journal: Chinese Journal of Dermatology Year: 2020 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Type of study: Prognostic study Language: Chinese Journal: Chinese Journal of Dermatology Year: 2020 Type: Article