Bletilla striata polysaccharides alleviate metabolic dysfunction-associated steatotic liver disease through enhancing hepatocyte RelA/ HNF1α signaling

Figure 3 Stimulation with sodium oleate/sodium palmitate induces steatosis, and impairs nuclear factor kappa B p65/hepatocyte nuclear factor-1 alpha signaling in AML-12 cells. AML-12 cells were stimulated in triplicate with control bovine serum albumin, 250/125 μmol/L sodium oleate/sodium palmitate for 48 hours. A: Oil red O staining of lipids; B: Intracellular triglycerides content; C: Cell counting kit-8 analysis of cell viability; D: Flow cytometry analysis of the frequency of apoptotic cells; E: Intracellular malondialdehyde levels; F: Reduced glutathione levels; G: Western blot analysis of the relative levels of lipid metabolism-related protein expression; H: Western blot analysis of the relative levels of nuclear factor kappa B p65 and hepatocyte nuclear factor-1 alpha expression; I: Western blot analysis of the relative levels of endoplasmic reticulum stress-related protein expression; J: Western blot analysis of the relative levels of apoptosis-, and necroptosis-related protein expression. Data are representative images or expressed as the mean ± SD of each group from three separate experiments. ^bP < 0.01. ¹P < 0.01 vs the respective control group. SO/SP: Sodium oleate/sodium palmitate; TG: Triglycerides; MDA: Malondialdehyde; GSH: Glutathione; APC: Allophycocyanin conjugate; MLKL: Mixed lineage kinase domain-like protein; SREBP1c: Cleaved sterol regulatory element-binding protein 1; TG: Triglycerides; MTP: Microsomal triglyceride transfer protein; MCAD: Medium-chain acyl-CoA dehydrogenase; GPX4: Glutathione peroxidase 4; HNF1α: Hepatocyte nuclear factor-1 alpha; GRP78: 78-kDa glucose-regulated protein; CHOP: CCAAT-enhancer-binding protein homologous protein; HRD1: 3-hydroxy-3-methyl glutaryl coenzyme A reductase degradation 1 homolog; ATF4: Activating transcription factor 4; RelA: Nuclear factor kappa B p65.