Κυριακή 8 Νοεμβρίου 2020

Effects and Mechanism of Berberine on the Dexamethasone-Induced Injury of Human Tendon Cells

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Objective. To investigate the effects of berberine (Berb) on dexamethasone- (Dex-) induced injury of human tendon cells and its potential mechanism. Methods. CCK-8 assay was used to explore the appropriate concentration of Dex-induced injury of tendon cells and the doses of Berb attenuates Dex cytotoxicity; cell wound healing assay was used to detect the effects of Berb and Dex on the migration ability of tendon cells; flow cytometry was used to measure cell apoptosis; DCF DA fluorescent probe was used to measure the ROS activity of cells. Western blotting was used to detect the expression of phenotype related factors including smooth muscle actin α (SMA-α), type I collagen (Col I), col III, apoptosis-related factors, caspase-3, cleaved caspase-3, caspase-9, cleaved caspase-9, and PI3K/AKT. Results. CCK -8 assay showed that 1–100 μM Dex significantly inhibited the proliferation of tendon cells in a concentration-dependent manner , where the inhibitory effect of 100 μM Dex was most significant , and the pretreatment of 150, 200 μM Berb could reverse those inhibitions (all ). Compared with the control group, Dex significantly inhibited cell migration , while Berb pretreatment could enhance cell migration . Flow cytometry and ROS assay showed that Dex could induce apoptosis and oxidative stress response of tendon cells (all ), while Berb could reverse those responses . Western blot showed that Dex could inhibit the expression of the col I and III as well as α-SMA (all ) and enhance the expression of apoptosis-related factors including cleaved caspase-3 and cleaved caspase-9 (all ). Besides, Dex could also inhibit the activation of the PI3K/AKT signaling pathway (all ), thus affecting cell function, while Berb treatment significantly reversed the expression of those above p roteins (all ).Conclusion. Berb attenuated DEX induced reduction of proliferation and migration, oxidative stress, and apoptosis of tendon cells by activating the PI3K/AKT signaling pathway and regulated the expression of phenotype related biomarkers in tendon cells. However, further studies are still needed to clarify the protective effects of Berb in vivo.
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