Effect Of Dihydroartemisinin On Fibroblasts Of Human Hypertrophic Scar And Its Mechanism

Purpose:Here, we investigated the effect of dihydroartemisinin on treating human hypertrophic scar fibroblast(hHSF) in vitro and tried to elucidate the possible mechanisms. Thus lay the theoretical foundation for the development of dihydroartemisinin for clinical use.Material and method:Cell viability was determined by MTT assay after treatment with dihydroartemisinin for 24h and 48h. Number of alive cell was determined by Trypan blue assay after treatment with dihydroartemisinin for 24h and 48h. Cellular migration abilities were assayed by scratch test. Apoptosis were determined by Annexin V-FITC/PI assay after treatment with dihydroartemisinin for 24h. Cell cycle kit was used to determin the change of cell cycle by flow cytometry analysis after treatment with dihydroartemisinin for 24h. Western blot analysis was performed to study the expression of TGF-β1, caspase3, Cyclin D1 and collagen I. Real time RT-PCR was performed to detect the change in TGF-β1 mRNA expression.Results:1.Dihydroartemisinin showed the obvious cytotoxicity after treating hHSF in dose-dependent manner and time-dependent manner(IC50 of 24h= 14.0μM, IC50 of 48h= 9.3μM).2.Dihydroartemisinin could remarkbly suppressed hHSF cell proliferation and cellular migration abilities in dose-dependent manner.3.Dihydroartemisinin could remarkbly increase cell apoptosis and the expression of cleaved-caspase3. Also, dihydroartemisinin decreased the expression of cyclin D1 and induced G0-G1 arrest in hHSF.4.Dihydroartemisinin decreased the expression of TGF-β1 and collagen I in dose-dependent manner.Conclusion:1.Dihydroartemisinin increases cell apoptosis by increasing the expression of cleaved-caspase3 and arrests the cell cycle by decreasing the expression of cyclin D1 leading to a obvious cytotoxicity and inhibition of cell proliferation in hHSF.2.Dihydroartemisinin suppresses cellular migration abilities of hHSF.3.Inhibition of collagen I expression caused by inhibition of TGF-J31 mRNA and protein expression may be the possible mechanisms of dihydroartemisinin effect.