Apoptosis

, Volume 19, Issue 1, pp 30–41

MicroRNA-29a protects against acute liver injury in a mouse model of obstructive jaundice via inhibition of the extrinsic apoptosis pathway

  • Mao-Meng Tiao
  • Feng-Sheng Wang
  • Li-Tung Huang
  • Jiin-Haur Chuang
  • Ho-Chang Kuo
  • Ya-Ling Yang
  • Ying-Hsien Huang
Original Paper

Abstract

Recent studies have shown that microRNA-29 (miR-29) is significantly decreased in liver fibrosis, as demonstrated in human liver cirrhosis, and that its downregulation influences the activation of hepatic stellate cells. In addition, both cleaved caspase-3 production and apoptosis play a role in cholestatic liver injury. However, it is unknown if miR-29 is effective in modulating the extent of injury. We employed miR-29a transgenic mice (miR-29aTg mice) and wild-type (WT) littermates to clarify the role of miR-29 in hepatic injury and fibrogenesis, using the bile duct-ligation (BDL) mouse model. After BDL, all three members of the miR-29 family were significantly downregulated in the livers of WT mice, and miR-29b and miR-29c were significantly downregulated in the livers of the miR-29aTg mice. Liver function, as measured by alanine transaminase and aspartate transaminase activity, was significantly improved in the miR-29aTg mice than in the WT littermates, following 1 week of obstructive jaundice. In addition, overexpression of miR-29a was associated with a significant downregulation of the expression of collagen-1α1, collagen-4α1, phospho-FADD, cleaved caspase-8, cleaved caspase-3, Bax, Bcl-2, PARP, and nuclear factor-κB, as well as an upregulation of phospho-AKT expression. In addition, there were significantly fewer TUNEL-positive liver cells in the miR-29aTg group than in the WT littermates after BDL. Our results indicate that miR-29a decreases cholestatic liver injury and fibrosis after BDL, at least partially, by modulating the extrinsic rather than intrinsic pathway of apoptosis.

Keywords

Caspase Mir-29 Cholestasis TUNEL 

Abbreviations

BDL

Bile duct ligation

ECM

Extracellular matrix

HSC

Hepatic stellate cells

LDH

Lactate dehydrogenase

TLR

Toll like receptor

TUNEL

TdT-mediated dUTP biotin nick-end labeling

WT

Wide type

XIAP

X-linked inhibitor of the apoptotic protein

Supplementary material

10495_2013_909_MOESM1_ESM.tif (77 kb)
Fig. S1Quantitative measurement of collagen-1α1 expression by immunofluorescence staining in liver tissue of miR-29a transgenic mice and WT littermates underwent sham operation and bile duct ligation (BDL). Data are expressed as mean ± standard error in 4 samples from miR-29a transgenic mice and 4 samples from WT littermates (TIFF 76 kb)
10495_2013_909_MOESM2_ESM.tif (1.9 mb)
Fig. S2Immunoreactive caspase 8 staining between miR-29aTg mice and wild type littermates. There was significantly higher caspase-8 immunoreactivity in the liver tissue of the sham-operated and BDL groups than in the control group, which was expressed constantly in the cytoplasm of hepatocytes (arrowhead). Moreover, overexpression of miR-29a decreased the expression of caspase-8 and caspase-3 after BDL (TIFF 1900 kb)
10495_2013_909_MOESM3_ESM.tif (595 kb)
Fig. S3Comparison of the X-linked inhibitor of the apoptotic protein (XIAP) expression by western blot in liver tissue of miR-29a transgenic mice and WT littermates that underwent sham operation and bile duct ligation (BDL). There were significantly downregulated XIAP protein expression in both miR-29aTg mice and WT littermates after BDL. However, overexpression did not change of XIAP protein expression after BDL. Data are expressed as mean ± standard error in 8 samples from miR-29a transgenic mice and 6 samples from WT littermates (TIFF 595 kb)

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Mao-Meng Tiao
    • 1
  • Feng-Sheng Wang
    • 2
  • Li-Tung Huang
    • 1
  • Jiin-Haur Chuang
    • 3
  • Ho-Chang Kuo
    • 1
  • Ya-Ling Yang
    • 4
  • Ying-Hsien Huang
    • 1
  1. 1.Department of PediatricsKaohsiung Chang Gung Memorial Hospital and the Graduate Institute of Clinical Medical Sciences, Chang Gung University College of MedicineKaohsiungTaiwan, ROC
  2. 2.Department of Medical ResearchKaohsiung Chang Gung Memorial Hospital and Chang Gung University College of MedicineKaohsiungTaiwan, ROC
  3. 3.Department of SurgeryKaohsiung Chang Gung Memorial Hospital and Chang Gung University College of MedicineKaohsiungTaiwan, ROC
  4. 4.Department of AnesthesiologyKaohsiung Chang Gung Memorial Hospital and Chang Gung University College of MedicineKaohsiungTaiwan, ROC

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