Salvianolic acid B protects hepatocytes from H₂O₂ injury by stabilizing the lysosomal membrane

To investigate the capability of salvianolic acid B (Sal B) to protect hepatocytes from hydrogen peroxide (H₂O₂)/carbon tetrachloride (CCl₄)-induced lysosomal membrane permeabilization.

Cell Counting Kit-8 assay was used to measure cell viability. Apoptosis and death were assayed through flow cytometry. BrdU incorporation was used to detect cell proliferation. Serum alanine aminotransferase activity and liver malondialdehyde (MDA) content were measured. Liver histopathological changes were evaluated using hematoxylin-eosin staining. Lysosomal membrane permeability was detected with LysoTracker Green-labeled probes and acridine orange staining. The levels of protein carbonyl content (PCC), cathepsins (Cat)B/D, and lysosome-associated membrane protein 1 (LAMP1) were evaluated through western blotting. Cytosol CatB activity analysis was performed with chemiluminescence detection. The mRNA level of LAMP1 was evaluated through quantitative real-time polymerase chain reaction.

Results indicated that H₂O₂ induced cell injury/death. Sal B attenuated H₂O₂-induced cell apoptosis and death, restored the inhibition of proliferation, decreased the amount of PCC, and stabilized the lysosome membrane by increasing the LAMP1 protein level and antagonizing CatB/D leakage into the cytosol. CCl₄ also triggered hepatocyte death. Furthermore, Sal B effectively rescued hepatocytes by increasing LAMP1 expression and by reducing lysosomal enzyme translocation to the cytosol.

Sal B protected mouse embryonic hepatocytes from H₂O₂/CCl₄-induced injury/death by stabilizing the lysosomal membrane.