HO-1 Is Essential for Tetrahydroxystilbene Glucoside Mediated Mitochondrial Biogenesis and Anti-Inflammation Process in LPS-Treated RAW264.7 Macrophages
TSG promoted the mitochondrial biogenesis in RAW264.7 macrophages. RAW264.7 cells were treated with TSG (0, 30, 60, and 120 μM) for 6 h. (a) Mito-tracker red CMX-Ros (red) and DAPI (blue) were used to stain the mitochondria mass and cell nucleus. Fluorescence images were captured by a confocal microscopy. Bar = 10 μm (×240). (b) Mitochondrial mass was also assessed by flow-cytometry method with Mito-tracker green staining. (c) The Mito-tracker green fluorescence was assessed by using Image-Pro plus 6 software, and mean fluorescence was calculated. (d) Western blot analysis was performed to assess the expression of mitochondrial complex IV in whole cell lysates. (e) Quantitative PCR was performed to evaluate the mtDNA content. Relative ratios of mtDNA and nDNA contents were analyzed. (f) The mRNA expressions of PGC-1, NRF1, and TFAM were detected by Real-Time PCR methods. All data were expressed as means ± SEM of triplicate independent experiments. and versus control group.