Abstract:
BACKGROUND:Intraarticular scar adhesion refers to a serious complication caused by knee surgery or trauma, leading to various sequelae (e.g., articular cartilage degeneration and knee joint stiffness). Artesunate (ART) has exhibited an effect to suppress fibroblast proliferation, whereas the exact mechanism remains unclear. This study aims to delve into the possible mechanism of ART in suppressing joint adhesion. METHODS:The effect of ART on reduced intraarticular adhesions was ascertained by histological staining and immunohistochemical analysis through vivo experiments. Cell Counting Kit-8 (CCK-8) assay, Western blot analysis, flow cytometry, and tunnel staining were used to detect the effect of ART in promoting fibroblast apoptosis and delve into its possible signaling pathway. RESULTS:The results of hematoxylin-eosin (HE) staining suggested that the number of fibroblasts decreased with the increase in ART concentration. The results of Masson staining were similar, with the increase in concentration, the collagen content decreased. Immunohistochemical results showed that the expression of endoplasmic reticulum stress (ERS) characteristic proteins 78 kDa glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) increased in a concentration-dependent manner. CCK-8 results suggested that ART could inhibit fibroblast viability in a concentration- and time-dependent manner. Results of flow cytometry, tunnel staining, and Western blot suggested the apoptosis of fibroblasts occurred after ART treatment. Cells with caspase inhibitors were treated, and apoptotic proteins cleaved-poly ADP-ribose polymerase (cleaved PARP) and cleaved-caspase 3 were detected; the results showed that the apoptotic effect of ART was reduced. The expressions of ERS-related protein CHOP and apoptosis-related protein Bax were upregulated, while the expression of Bcl-2 was downregulated, and the ratio of Bax/Bcl-2 increased in a concentration-dependent manner. Continuous detection of PRKR-like ER kinase (PERK) pathway-related proteins showed that the expression of p-PERK and phosphorylating eukaryotic initiation factor 2α (p-eIF2α) increased in a time-dependent and concentration-dependent manner. PERK pathway inhibitors could partially inhibit ART-mediated apoptosis through PERK pathway. CONCLUSIONS:ART can promote fibroblast apoptosis through PERK pathway, a classical ERS pathway, and thus prevent fibrosis in the surgical area after joint surgery.
journal_name
J Orthop Surg Resjournal_title
Journal of orthopaedic surgery and researchauthors
Chen H,Tao J,Wang J,Yan Ldoi
10.1186/s13018-019-1445-xsubject
Has Abstractpub_date
2019-12-17 00:00:00pages
448issue
1issn
1749-799Xpii
10.1186/s13018-019-1445-xjournal_volume
14pub_type
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