ABSTRACT
Objective To investigate the effect of atractylone on the viability and apoptosis of hepatoma HepG2 cells and its mechanism of action. Methods Hepatoma HepG2 cells were selected and divided into low-, middle-, and high-dose atractylone groups (5, 10, and 20 μmol/L), and the cells in the control group were added with an equal volume of DMSO. MTT colorimetry was used to measure the viability of HepG2 cells after treatment with different concentrations of atractylone; flow cytometry was used to measure the apoptosis rate and mitochondrial membrane potential of HepG2 cells; the DCFH-DA fluorescent probe labeling method was used to measure the level of reactive oxygen species (ROS) in HepG2 cells; Transwell assay was used to evaluate the effect of atractylone on the migration ability of HepG2 cells; Western blot was used to measure the protein expression levels of Bcl-2, Bax, and cleaved caspase-3. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for comparison between two groups. Results After 24 and 48 hours of treatment with atractylone, compared with the control group, the low-, middle-, and high-dose atractylone groups had a tendency of reduction in cell viability (all P < 0.05), with a half inhibitory concentration of 26.19 μmol/L in atractylone treatment of HepG2 cells for 72 hours. The low-, middle-, and high-dose atractylone groups had a significantly higher apoptosis rate than the control group (14.34%/29.32%/50.12% vs 0.32%, all P < 0.05). Compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant increase in the fluorescence intensity of ROS in HepG2 cells (all P < 0.05). After 48 hours of treatment with atractylone, compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant reduction in the number of migrated cells (132.67±18.36/57.00±9.26/31.00±2.45 vs 258.11±38.54, P < 0.05). Compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant reduction in the expression of the anti-apoptotic factor Bcl-2 and significant increases in the expression of the apoptotic factors Bax and cleaved caspase-3 (all P < 0.05). Conclusion Atractylone can induce the apoptosis and inhibit the migration of HepG2 cells, which provides an experimental basis for further development and utilization of atractylone.
ABSTRACT
Objective To investigate the effect of miRNA-384 (miR-384)expression on hepatic steatosis in mice with nonalcoholic fatty liver disease (NAFLD)induced by high-fat diet (HFD). Methods A total of 30 male C57BL/6J mice were fed for 7 days to adapt to the environment and then randomly divided into 2 groups,with 15 mice in each group. The mice in the control group were given normal diet, and those in the model group were given HFD for 8 weeks and then the liver tissue was harvested. HE and Nile red staining were used to ob-serve the pathological changes of the liver. Microarray sequencing was performed to determine the whole-genome miRNA expression profile of liver tissue,and PCR was used to measure the relative expression of miR-384. The t-test was used for the comparison of continuous da-ta between groups. Results In the control group,the liver was red with sharp edges,the lobular structure was clear,and there was no he-patic steatosis;in the model group,the liver was yellow with blunt edges,and the hepatocytes were swollen with a large number of fat vacu-oles in the cytoplasm and nuclear deviation caused by the fusion of lipid droplets. Compared with the normal mice,the NAFLD mice had 12 upregulated miRNAs and 18 downregulated miRNAs in liver tissue. Some of the differentially expressed miRNAs between the control group and the model group were screened to obtain the same cluster diagram. Among the 8 miRNAs with significant changes,miR-384 showed a significant fold change. Conclusion The upregulation of miR -384 is closely associated with hepatic steatosis,but its mechanism still needs further study.