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

Figure 4 Treatment with Bletilla striata polysaccharides is relatively safe in healthy mice. All mice were randomized into five groups (12 individuals each) including the normal control group (untreated); solvent control group (control; phosphate buffer saline: 20 mL/kg; quaque die; irrigation; 4 weeks); simple Bletilla striata polysaccharides group (0.1, 0.2, 0.4 g/kg; quaque die; irrigation; 4 weeks) (n = 12). Twelve hours after the final treatment, the mice were fasted for 6 h and their blood and liver samples were collected. A: Liver weights; B: Serum alanine aminotransferase levels; C: Serum total cholesterol levels; D: Serum triglyceride levels; E: Liver tissue triglyceride contents; F: Apoptotic index; G: Hematoxylin and eosin staining analysis of liver tissue pathology and transferase-mediated deoxyuridine triphosphate-nick end labeling analysis of liver cell apoptosis; H: Hepatic malondialdehyde levels; I: Hepatic reduced glutathione levels; J: Western blot analysis of the relative levels of hepatic lipid metabolism-related protein expression; K: Western blot analysis of relative hepatic nuclear factor kappa B p65, and hepatocyte nuclear factor-1 alpha protein levels; L: Western blot analysis of hepatic endoplasmic reticulum stress-related protein expression; M: Western blot analysis of hepatic cleaved caspase-3 and mixed lineage kinase domain-like protein phosphorylation. Data are representative images or expressed as the mean ± SD of each group of mice from at least three separate experiments. ^bP < 0.01. ¹P < 0.01 vs the untreated group. NC: Normal control; ALT: Alanine aminotransferase; TC: Total cholesterol; TG: Triglycerides; BSP: Bletilla striata polysaccharides; HE: Hematoxylin and eosin; MLKL: Mixed lineage kinase domain-like protein; TUNEL: Transferase-mediated deoxyuridine triphosphate-nick end labeling; wk: Week; MDA: Malondialdehyde; GSH: Glutathione; PBS: Phosphate buffer saline; SREBP1c: Cleaved sterol regulatory element-binding protein 1; 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.