ABSTRACT
The aim of this study was to investigate the role and mechanism of terpinen-4-ol (T4O) on high glucose (HG) -induced calcification in vascular smooth muscle cell (VSMC). To investigate the role of T4O on HG-induced calcium deposition, osteogenic phenotypic transformation and mitochondrial dynamics in VSMC, Mdivi-1, a mitochondrial dynamin-related protein 1 (Drp-1) inhibitor, was used to analyze the correlation between mitochondrial dynamics and VSMC calcification and the role of T4O. Alizarin red S staining was used to observe calcium salt deposition and flow cytometry to detect intracellular Ca2+ content; Western blot and immunofluorescence were used to detect the expression of phenotypic switching-related markers α-smooth muscle actin (α-SMA), bone morphogenetic protein 2 (BMP2) and Runt related transcription factor 2 (Runx2), and mitochondrial dynamics-related markers mitofusin 1 (MFN1), mitofusin 2 (MFN2) and Drp-1. The results showed that low and high doses of T4O could inhibit HG-induced down-regulation of α-SMA, MFN1 and MFN2 expression levels, and up-regulation of BMP2, Runx2 and Drp-1 expression levels, reduce intracellular Ca2+ content and calcium salt deposition, and effectively inhibit HG-induced VSMC calcification and mitochondrial dynamics disorders. The T4O group, Mdivi-1 group and T4O+Mdivi-1 group were able to up-regulate the expression levels of HG-induced α-SMA, MFN1 and MFN2, down-regulate the protein expression levels of BMP2, Runx2 and Drp-1, and inhibit calcium salt deposition, and there was no significant difference between the above indexes in the T4O and T4O+Mdivi-1 groups. The above findings suggest that T4O can inhibit the expression level of Drp-1, regulate the disturbance of mitochondrial dynamics, and suppress HG-induced VSMC calcification.
ABSTRACT
Objective To predict and analyze the structure and function of Stenotrophomonas maltophilia OmpA. Methods Bioinformatics software and biological databases were used to analyze the physicochemical properties, signal peptides,and transmembrane structures of the OmpA protein and to predict the subcellular localization, secondary and tertiary structures,sequence homology and conserved domains,and epitopes of OmpA.Results OmpA protein had strong hydrophilicity but without transmembrane helices in mature protein,and positions 1-22 of the sequence were predicted as signal peptide.In the second structure random coil helix,αlpha-helix,beta-turn and extended strand made up 50.27%, 24.59%,9.29%and 15.85%,respectively.The three-dimensional structure was β-barrel.OmpA was highly conserved among S.maltophilia strains but shared minimal homology with human and mouse proteins.The N-terminal domain of OmpA was OM-channels superfamily and the C-terminal domain of OmpA was OmpA_C-like superfamily.OmpA protein contained 11 dominant antigen epitopes.Conclusion The characteristics of OmpA identified by bioinformatics analysis can not only provide reference for the study of the structure and novel function of OmpA,but also theoretically contribute to the research on related new subunit vaccines.