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Stage-specific expression, immunolocalization of Clonorchis sinensis lysophospholipase and its potential role in hepatic fibrosis

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Abstract

Lysophospholipase, belonging to the complex family of phospholipases, is supposed to play a vital role in virulence and pathogenesis of parasites and fungi. In the current study, the potential role of Clonorchis sinensis lysophospholipase (CslysoPLA) in hepatic fibrosis induced by C. sinensis was explored for the first time. In the liver of the cat infected with C. sinensis, CslysoPLA was recognized in the lumen between adult worms and surrounding bile duct epithelia together with some inside the cells by means of immunolocalization. Both Cell Counting Kit-8 (CCK-8 assay) and cell cycle analysis of human hepatic stellate cell line LX-2 showed that a higher percentage of cells were at proliferation phase after incubation with lower concentrations of recombinant CslysoPLA (rCslysoPLA). Quantitative real-time polymerase chain reaction (RT-PCR) demonstrated an upregulation in fibrogenic genes of smooth muscle α-actin, collagen III, matrix metalloproteinase 2 and tissue inhibitors of metalloproteinase II in LX-2 treated with rCslysoPLA. Moreover, human biliary epithelial cell line 5100 proliferated significantly in response to rCslysoPLA. Notably, CslysoPLA was localized in the adenomatoid hyperplastic tissue within the intrahepatic bile duct of experimentally infected rats by immunolocalization analysis. In addition, quantitative RT-PCR implied that CslysoPLA was differentially expressed at the developmental stages of C. sinensis (metacercariae, adult worms and eggs), with the highest level at metacercariae stage. Immunolocalization analysis showed that CslysoPLA was distributed in the intestine, vitelline gland, tegument and eggs in the adult worms and in the tegument and vitelline gland in the metacercariae, respectively. Collectively, it suggests that CslysoPLA might be involved in the initiation and promotion of C. sinensis-related human hepatic fibrosis and advance future studies on its promotion to C. sinensis-induced cholangiocarcinogenesis.

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Acknowledgments

This work was supported by the National Key Basic Research and Development Project of China (973 project; no. 2010CB530000), National Natural Science Foundation of China (no. 81101270 and no. 81171602), China Postdoctoral Science Foundation (no. 20110490952), Fundamental Research Funds for the Central Universities (no. 3164015) and the National Important Sci-tech Special Projects (no. 2012ZX10004220).

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Authors and Affiliations

  1. Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, 510080, People’s Republic of China

    Fan Zhang, Pei Liang, Wenjun Chen, Xiaoyun Wang, Yue Hu, Chi Liang, Jiufeng Sun, Yan Huang, Ran Li, Xuerong Li, Jin Xu & Xinbing Yu

  2. Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-sen University, Guangzhou, 510080, People’s Republic of China

    Fan Zhang, Pei Liang, Wenjun Chen, Xiaoyun Wang, Yue Hu, Chi Liang, Jiufeng Sun, Yan Huang, Ran Li, Xuerong Li, Jin Xu & Xinbing Yu

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  1. Fan Zhang
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  9. Ran Li
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  12. Xinbing Yu
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Correspondence to Xinbing Yu.

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Fan Zhang and Pei Liang have contributed equally to this work.

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Zhang, F., Liang, P., Chen, W. et al. Stage-specific expression, immunolocalization of Clonorchis sinensis lysophospholipase and its potential role in hepatic fibrosis. Parasitol Res 112, 737–749 (2013). https://doi.org/10.1007/s00436-012-3194-1

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  • DOI: https://doi.org/10.1007/s00436-012-3194-1

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