Effect of oxaliplatin on the activation of hepatic stellate cells and its mechanism / 临床肝胆病杂志
Journal of Clinical Hepatology
; (12): 1142-1148, 2024.
Article
em Zh
| WPRIM
| ID: wpr-1032262
Biblioteca responsável:
WPRO
ABSTRACT
ObjectiveTo investigate the effect of oxaliplatin on the activation of hepatic stellate cells (HSCs), as well as the association of oxaliplatin with microRNA-30a-5p and autophagy. MethodsHSC-LX2 cells were cultured and divided into groups according to the following three protocols: control group, PDGF treatment group, oxaliplatin treatment group, oxaliplatin+PDGF treatment group; control group, microRNA-30a-5p transfection group, PDGF treatment group, microRNA-30a-5p transfection+PDGF treatment group; control group, 3-MA group, microRNA-30a-5p inhibitor group, microRNA-30a-5p inhibitor+3-MA group. Western Blot was used to measure the expression of HSC activation-related proteins (Collagen-I and alpha-smooth muscle actin [α- SMA]) and HSC autophagy-related proteins (Beclin-1, P62, and LC3B); LysoTracker staining and immunofluorescence assay were used to measure the expression of LC3B autophagosomes; RT-PCR was used to measure the expression level of microRNA-30a-5p; bioinformatics techniques were used to predict the potential targets of microRNA-30a-5p in HSCs. The independent-samples t test was used for comparison of normally distributed continuous data between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsAfter the cells were treated with oxaliplatin, RT-PCR results showed that the oxaliplatin treatment group had a significantly higher expression level of microRNA-30a-5p than the control group (P<0.01); Western Blot showed that the oxaliplatin treatment group had significant reductions in the expression levels of the HSC activation-related proteins α-SMA and Collagen-Ⅰ and the autophagy-related proteins Beclin 1 and LC3BⅡ/Ⅰ (all P<0.001); immunofluorescence assay showed that the oxaliplatin treatment group had a significantly lower number of autophagosomes than the control group (P<0.05). After HSC-LX2 cells were transfected with microRNA-30a-5p mimic, compared with the control group, the microRNA-30a-5p mimic group had significant reductions in the expression levels of the autophagy-related proteins Beclin 1 and LC3BⅡ/Ⅰ (P<0.05) and the HSC activation-related protein Collagen-Ⅰ (P<0.001); after HSC-LX2 cells were transfected with microRNA-30a-5p inhibitor, Western Blot showed that compared with the control group, the microRNA-30a-5p inhibitor group had significant increases in the expression levels of the HSC activation-related proteins Collagen-Ⅰ and α-SMA and the autophagy-related protein Beclin 1 (t=2.41, 2.32, and 4.57, all P<0.05). Western Blot showed that compared with the control group, the microRNA-30a-5p inhibitor group had significant increases in the expression levels of the HSC autophagy-related protein Beclin 1 and the HSC activation-related protein α-SMA (both P<0.05), and after the treatment with the autophagy inhibitor 3-MA, there were no significant differences in the expression of these proteins between the two groups (P>0.05). The bioinformatics analysis using TargetScan, PicTar, and miRanda databases showed that the autophagy-related protein Beclin-1 might be a potential target of miRNA-30a-5p. ConclusionOxaliplatin can inhibit the activation of HSCs by upregulating the expression of microRNA-30a-5p, which provides new ideas and a new target for the treatment of liver fibrosis.
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Índice:
WPRIM
Idioma:
Zh
Revista:
Journal of Clinical Hepatology
Ano de publicação:
2024
Tipo de documento:
Article