ABSTRACT
Objective To study the effect and mechanism of blueberry on regulating the mitochondrial inner membrane protein mitofilin/Mic60 in an in vitro model of metabolic dysfunction-associated liver disease (MAFLD). Methods L02 human hepatocytes were induced by free fatty acids (FFA) to establish MAFLD cell model. A normal group, a model group, an 80 μg/mL blueberry treatment group, a Mic60 short hairpin RNA (Mic60 shRNA) transfection group, and Mic60 knockdown combined with an 80 μg/mL blueberry treatment group were established. The intracellular lipid deposition was observed by oil red O staining, and the effect of different concentrations of blueberry pulp on the survival rate of L02 cells treated with FFA was measured by MTT assay. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), total cholesterol (TC), superoxide dismutase (SOD) activity, glutathione (GSH) and malondialdehyde (MDA) contents were measured by visible spectrophotometry. The expression of reactive oxygen species (ROS) in hepatocytes was observed by fluorescence microscopy, and the mRNA and protein expression of Mic60 were detected by real-time quantitative PCR and Western blot analysis, respectively. Results After 24 hours of FFA stimulation, a large number of red lipid droplets in the cytoplasm of L02 cells was observed, and the survival rate of L02 cells treated with 80 μg/mL blueberry was higher. The results of ALT, AST, TG, TC, MDA and the fluorescence intensity of ROS in blueberry treated group were lower than those in model group, while the levels of SOD, GSH, Mic60 mRNA and protein in blueberry treated group were higher than those in model group. Conclusion Blueberry promotes the expression of Mic60, increases the levels of SOD and GSH in hepatocytes, and reduces the production of ROS, thus alleviating the injury of MAFLD hepatocytes and regulating the disorder of lipid metabolism.
Subject(s)
Humans , Blueberry Plants/chemistry , Hepatocytes/metabolism , Liver/metabolism , Liver Diseases/metabolism , Reactive Oxygen Species/metabolism , Superoxide Dismutase/metabolism , Superoxides/metabolism , Mitochondrial Membranes/metabolism , Mitochondrial Proteins/metabolism , Plant Extracts/pharmacologyABSTRACT
OBJECTIVE:To establish HPLC method in the determination of the related substances in cefalotin sodium.METHODS:The determination was performed with C 18 as the chromatographic column.The mobile phase was consisted of sodium acetate solution-acetonitrile(82∶18)with a flow rate at1.0ml/min.The column temperature was30℃and the detection wavelength was254nm.RESULTS:All the chief peaks of cefalotin sodium and those of their related substances could be well isolated in all the specificity tests.The total peak area of the chief foreign substances of all the3batches of samples were no more than the twice of the chief peak area of their control solutions.CONCLUSION:This method is simple,accurate and reproducible.