The Imbalance of Mitochondrial Homeostasis of Peripheral Blood-Derived Macrophages Mediated by MAFLD May Impair the Walking Ability of Elderly Patients with Osteopenia

Figure 5

MAFLD impaired aerobic respiration and mitochondrial morphology of CD14+ monocytes in peripheral blood of patients with osteopenia. (a) The protein level of BAX, BLC2, and CYCS in Thp-1 induced by PMA and CD14 + momocytes in human peripheral blood is detected by WB. (b) In the heat map, the relative mRNA level of TLR4, BAX/BCL-2, CYCS, and Myd88 in PBMCs from OPe, OPe with MAFLD, and SO patients is determined by RT-PCR. (c) Representative image from immunofluorescence staining assay of COXIV and CYCS in THP-1 induced by PMA and the supernatant of HepG2 fatty liver model. COXIV (green), CYCS (red). (d) The OCR and ECAR in CD14 + PBMCs from the patients among OPe, OPe with MAFLD, and SO are detected by Seahorse XF. The bar plots indicate the quantitation of non-mitochondrial respiration, basal respiration, respiration capacity, and fatty acid oxidation among OPe, OPe with MAFLD, and SO. (e) The mitochondrial microstructure and morphology of CD14+ PBMCs from OPe and OPe with MAFLD patients are observed via transmission electron microscopy. (f) The level of mitochondrial ROS and intracellular ROS is assessed by FCA among OPe, OPe with MAFLD, and SO. The quantitative results showed right. Bars, means SE; , ; , ; , ; , . Abbreviations: TLR4: toll-like receptor 4; MyD88: myeloid differentiation factor 88; BCL-2: B-cell lymphoma-2; BAX: BCL2-associated X protein; CYCS: Cytochrome C; COXIV: Cytochrome c oxidase subunit IV; ROS: reactive oxygen species; PBMC: peripheral blood monocytes cell; SO: severe osteoporosis; OPe: osteopenia; OPe with MAFLD: osteopenia with metabolic dysfunction-associated fatty liver disease.

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