title: Stage-specific expression, immunolocalization of Clonorchis sinensis lysophospholipase and its potential role in hepatic fibrosis.
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.
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China.
authors.forename: Fan
authors.initials: F
authors.lastname: Zhang
authors.forename: Pei
authors.initials: P
authors.lastname: Liang
authors.forename: Wenjun
authors.initials: W
authors.lastname: Chen
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Wang
authors.forename: Yue
authors.initials: Y
authors.lastname: Hu
authors.forename: Chi
authors.initials: C
authors.lastname: Liang
authors.forename: Jiufeng
authors.initials: J
authors.lastname: Sun
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.forename: Ran
authors.initials: R
authors.lastname: Li
authors.forename: Xuerong
authors.initials: X
authors.lastname: Li
authors.forename: Jin
authors.initials: J
authors.lastname: Xu
authors.forename: Xinbing
authors.initials: X
authors.lastname: Yu
mesh_terms: D000818:Animals; D002415:Cats; D002460:Cell Line; D049109:Cell Proliferation; D003003:Clonorchiasis / Q000469:parasitology* / Q000473:pathology; D003004:Clonorchis sinensis / Q000201:enzymology* / Q000472:pathogenicity*; D004195:Disease Models, Animal; D020869:Gene Expression Profiling; D006801:Humans; D008103:Liver Cirrhosis / Q000469:parasitology / Q000473:pathology*; D008245:Lysophospholipase / Q000096:biosynthesis*; D051381:Rats
chemical_list: D008245:Lysophospholipase