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Activation of Mir-29a in Activated Hepatic Stellate Cells Modulates Its Profibrogenic Phenotype through Inhibition of Histone Deacetylases 4.

Huang YH, Tiao MM, Huang LT, Chuang JH, Kuo KC, Yang YL, Wang FS - PLoS ONE (2015)

Bottom Line: In this study, we examined whether miR-29a was involved in the regulation of HDAC4 and modulation of the profibrogenic phenotype in HSCs.Overexpression of miR-29a and HDAC4 RNA-interference decreased the expression of fibrotic genes, HDAC4 signaling, and HSC migration and proliferation.In contrast, knockdown of miR-29a with an antisense inhibitor increased HDAC4 function, restored HSC migration, and accelerated HSC proliferation.

View Article: PubMed Central - PubMed

Affiliation: Departments of Pediatrics, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan.

ABSTRACT

Background: Recent studies have shown that microRNA-29 (miR-29) is significantly decreased in liver fibrosis and that its downregulation influences the activation of hepatic stellate cells (HSCs). In addition, inhibition of the activity of histone deacetylases 4 (HDAC4) has been shown to strongly reduce HSC activation in the context of liver fibrosis.

Objectives: In this study, we examined whether miR-29a was involved in the regulation of HDAC4 and modulation of the profibrogenic phenotype in HSCs.

Methods: We employed miR-29a transgenic mice (miR-29aTg mice) and wild-type littermates to clarify the role of miR-29a in cholestatic liver fibrosis, using the bile duct-ligation (BDL) mouse model. Primary HSCs from both mice were treated with a miR-29a mimic and antisense inhibitor in order to analyze changes in profibrogenic gene expression and HSC activation using real-time quantitative RT-PCR, immunofluorescence staining, western blotting, and cell proliferation and migration assays.

Results: After BDL, overexpression of miR-29a decreased collagen-1α1, HDAC4 and activated HSC markers of glial fibrillary acidic protein expression in miR-29aTg mice compared to wild-type littermates. Overexpression of miR-29a and HDAC4 RNA-interference decreased the expression of fibrotic genes, HDAC4 signaling, and HSC migration and proliferation. In contrast, knockdown of miR-29a with an antisense inhibitor increased HDAC4 function, restored HSC migration, and accelerated HSC proliferation.

Conclusions: Our results indicate that miR-29a ameliorates cholestatic liver fibrosis after BDL, at least partially, by modulating the profibrogenic phenotype of HSCs through inhibition of HDAC4 function.

No MeSH data available.


Related in: MedlinePlus

Migration of primary activated HSCs was measured using a wound healing assay.A miR-29a mimic or HDAC4 RNAi significantly inhibited the migration of primary HSCs of WT mice. Data are expressed as the mean ± SE of four independent experiments. *indicates a p < 0.05 between the groups.
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pone.0136453.g008: Migration of primary activated HSCs was measured using a wound healing assay.A miR-29a mimic or HDAC4 RNAi significantly inhibited the migration of primary HSCs of WT mice. Data are expressed as the mean ± SE of four independent experiments. *indicates a p < 0.05 between the groups.

Mentions: In order to assess whether miR-29a may regulate HSC migration, a wound-healing assay was performed using primary HSCs. The results of the present study showed that a miR-29a mimic or HDAC4 RNAi inhibited the migration of primary HSCs (both p < 0.001; Fig 8). We then conducted a cell proliferation assay to examine the effects of miR-29a on cell proliferation. As showed in Fig 9, a miR-29a mimic or HDAC4 RNAi inhibited the proliferation of primary HSCs (both p < 0.001). In addition, addition of a miR-29a anti-sense inhibitor significantly increased HSC proliferation (p < 0.001).


Activation of Mir-29a in Activated Hepatic Stellate Cells Modulates Its Profibrogenic Phenotype through Inhibition of Histone Deacetylases 4.

Huang YH, Tiao MM, Huang LT, Chuang JH, Kuo KC, Yang YL, Wang FS - PLoS ONE (2015)

Migration of primary activated HSCs was measured using a wound healing assay.A miR-29a mimic or HDAC4 RNAi significantly inhibited the migration of primary HSCs of WT mice. Data are expressed as the mean ± SE of four independent experiments. *indicates a p < 0.05 between the groups.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4549141&req=5

pone.0136453.g008: Migration of primary activated HSCs was measured using a wound healing assay.A miR-29a mimic or HDAC4 RNAi significantly inhibited the migration of primary HSCs of WT mice. Data are expressed as the mean ± SE of four independent experiments. *indicates a p < 0.05 between the groups.
Mentions: In order to assess whether miR-29a may regulate HSC migration, a wound-healing assay was performed using primary HSCs. The results of the present study showed that a miR-29a mimic or HDAC4 RNAi inhibited the migration of primary HSCs (both p < 0.001; Fig 8). We then conducted a cell proliferation assay to examine the effects of miR-29a on cell proliferation. As showed in Fig 9, a miR-29a mimic or HDAC4 RNAi inhibited the proliferation of primary HSCs (both p < 0.001). In addition, addition of a miR-29a anti-sense inhibitor significantly increased HSC proliferation (p < 0.001).

Bottom Line: In this study, we examined whether miR-29a was involved in the regulation of HDAC4 and modulation of the profibrogenic phenotype in HSCs.Overexpression of miR-29a and HDAC4 RNA-interference decreased the expression of fibrotic genes, HDAC4 signaling, and HSC migration and proliferation.In contrast, knockdown of miR-29a with an antisense inhibitor increased HDAC4 function, restored HSC migration, and accelerated HSC proliferation.

View Article: PubMed Central - PubMed

Affiliation: Departments of Pediatrics, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan.

ABSTRACT

Background: Recent studies have shown that microRNA-29 (miR-29) is significantly decreased in liver fibrosis and that its downregulation influences the activation of hepatic stellate cells (HSCs). In addition, inhibition of the activity of histone deacetylases 4 (HDAC4) has been shown to strongly reduce HSC activation in the context of liver fibrosis.

Objectives: In this study, we examined whether miR-29a was involved in the regulation of HDAC4 and modulation of the profibrogenic phenotype in HSCs.

Methods: We employed miR-29a transgenic mice (miR-29aTg mice) and wild-type littermates to clarify the role of miR-29a in cholestatic liver fibrosis, using the bile duct-ligation (BDL) mouse model. Primary HSCs from both mice were treated with a miR-29a mimic and antisense inhibitor in order to analyze changes in profibrogenic gene expression and HSC activation using real-time quantitative RT-PCR, immunofluorescence staining, western blotting, and cell proliferation and migration assays.

Results: After BDL, overexpression of miR-29a decreased collagen-1α1, HDAC4 and activated HSC markers of glial fibrillary acidic protein expression in miR-29aTg mice compared to wild-type littermates. Overexpression of miR-29a and HDAC4 RNA-interference decreased the expression of fibrotic genes, HDAC4 signaling, and HSC migration and proliferation. In contrast, knockdown of miR-29a with an antisense inhibitor increased HDAC4 function, restored HSC migration, and accelerated HSC proliferation.

Conclusions: Our results indicate that miR-29a ameliorates cholestatic liver fibrosis after BDL, at least partially, by modulating the profibrogenic phenotype of HSCs through inhibition of HDAC4 function.

No MeSH data available.


Related in: MedlinePlus