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Osteopontin Contributes to TGF-β1 Mediated Hepatic Stellate Cell Activation

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Abstract

Background and Purpose

Liver fibrosis is characterized by accumulation of extracellular matrix. Our previous study found that osteopontin (OPN) increased in plasma of cirrhotic patients and indicative of cirrhosis staging. The present study was designed to investigate the expression of OPN in liver tissues and plasma of cirrhotic patients and further explore the role of OPN in human hepatic stellate cell (HSC) activation.

Methods

We used immunohistochemical staining and enzyme-linked immunosorbent assay to evaluate the expression level of OPN in liver tissues and plasma from cirrhotic patients, respectively. We produced lentivirus particles and infected target cell to manipulate OPN expression. Infection efficiency was determined by real-time RT-PCR and western blot. Cell proliferation was determined using CCK8 assay, and phenotypes of HSC activation were determined by real-time RT-PCR. OPN promoter activity was determined by dual luciferase reporter assay.

Results

We found that OPN expression in human cirrhotic liver tissues was upregulated compared to normal controls. In addition, its expression correlated with Child-Pugh classification, MELD score and the occurrence of complications. We further explored OPN level in patients’ plasma and showed that its level correlated with transforming growth factor-β1 (TGF-β1). In human HSC cell line LX-2, we found that change of OPN expression level could not only affect the proliferation of cells but also the TGF-β1 mediated HSC activation. Moreover, OPN was increased by TGF-β1 stimulation and regulated by TGF-β1 at transcription level.

Conclusions

OPN is upregulated in liver tissues and plasma of cirrhotic patients and promotes TGF-β1 mediated HSC activation.

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Acknowledgments

This work was supported by National Natural Sciences Foundation of China, No. 81070350. We thank Technician Zheng Chen in our lab and Dr. Jiuxu Bai from Shenyang General Hospital of PLA for technical assistance. We thank Professor Gaoliang Ouyang (School of Life Sciences, **amen University) for providing the OPN plasmid. We also thank Prof. Scott L. Friedman (Mount Sinai School of Medicine, USA) for providing LX-2 cells.

Conflict of interest

The authors declare no conflicts of interest related to this work.

Author information

Authors and Affiliations

  1. State Key Laboratory of Cancer Biology, ****g Hospital of Digestive Diseases, The Fourth Military Medical University, **’an, 710032, Shaanxi Province, People’s Republic of China

    **ao **ao, Yi Gang, Yong Gu, **dong Chu, **feng Zhou, **qiang Cai, Hui Zhang, Li Xu, Yongzhan Nie, Kaichun Wu, Zhiguo Liu & Daiming Fan

  2. Department of Digestive System, Shaanxi Provincial Crops Hospital of Chinese People’s Armed Police Force, **’an, 710054, Shaanxi Province, People’s Republic of China

    Yong Gu

  3. Department of Radiation Oncology, ****g Hospital, The Fourth Military Medical University, **’an, 710032, Shaanxi Province, People’s Republic of China

    Lina Zhao

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  1. **ao **ao
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Corresponding authors

Correspondence to Zhiguo Liu or Daiming Fan.

Additional information

**ao **ao and Yi Gang contributed equally to this work.

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10620_2012_2248_MOESM1_ESM.tif

Fig. S1 Comparison of OPN and TIMP1 expression levels in consecutive sections of cirrhotic liver tissues. Representative images of coordinate expression of OPN and TIMP1 (kappa = 0.518, P = 0.000) were showed (original magnification, 200×). (A, B) Positive OPN staining (A) and positive TIMP1 staining (B) were observed in consecutive sections of cirrhotic liver tissues. (C, D) Negative OPN staining (C) and negative TIMP1 staining (D) were observed in consecutive sections of cirrhotic liver tissues. Supplementary material 1 (TIFF 10,280 kb)

Supplementary material 2 (DOC 55 kb)

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**ao, X., Gang, Y., Gu, Y. et al. Osteopontin Contributes to TGF-β1 Mediated Hepatic Stellate Cell Activation. Dig Dis Sci 57, 2883–2891 (2012). https://doi.org/10.1007/s10620-012-2248-7

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