Objective To investigate effects and mechanism of total glucosides of paeony on the proliferation, invasion, migration of HaCaT cells and the release of exosomes.
Methods HaCaT cells were treated with total glucosides of paeony at concentrations of 50, 100 and 200 μg/L, and were set as low dose group, medium dose group and high dose group, and cells added equal amount of complete culture medium was designed as control group. The proliferation ability was detected by methyl thiazolyl tetrazolium (MTT) method at different time points; the migration and invasion ability of cells were detected by Transwell chamber test; the exosomes were extracted by high-speed centrifugation; protein expressions of CyclinD1, protein kinase B (Akt), phosphorylated-Akt(p-Akt), phosphoinositide 3-kinase (PI3K), CD63 and CD81 were detected by western blot; the expression of CyclinD1, Akt, p-Akt and PI3K mRNA was detected by quantitative real-time PCR (qRT-PCR).
Results The inhibition rate of cell proliferation showed no difference among four groups at 0 h and 24 h (P>0.05). At 48 and 72 h of culture, the inhibition rates of cell proliferation were the highest in the high dose group, followed by medium dose group, low dose group, and were the lowest in the control group (P < 0.05). The high dose group had the largest number of migrating cells and invading cells, followed by medium dose group and low dose group, and the control group had the lowest(P < 0.05). Protein expressions of CyclinD1, CD63, CD81, Akt, p-Akt, PI3K and p-PI3K as well as expressions of CyclinD1, Akt, p-Akt, PI3K and p-PI3K mRNA were the highest in the high dose group, followed by medium dose group, low dose group, and were the lowest in the control group (P < 0.05).
Conclusion Glucosides of paeony can inhibit the secretion of exosomes, and the proliferation, invasion and migration of HaCaT cells. Its mechanism may be related to down-regulation of PI3K/Akt signaling pathway.
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