Bao Zheng1, Zhiyan Gao2, Liumei Liang3, Yunyu Lu1, Yongting Kong3, Wanting Chen3, Keying Lin3, Wanqi Chen3, Jingying Mai4, Yanwen Li5, Changling Ma6. 1. Department of Parasitology, School of Basic Medical Sciences, Guangxi Medical University, Nanning, 530021, People's Republic of China. 2. Department of Morphology Experiment Center, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, 511436, People's Republic of China. 3. KingMed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou, 511436, People's Republic of China. 4. Department of Pathogen Biology & Immunology, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, 511436, People's Republic of China. 5. Department of Parasitology, School of Basic Medical Sciences, Guangxi Medical University, Nanning, 530021, People's Republic of China. yanwenligx@aliyun.com. 6. Department of Pathogen Biology & Immunology, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, 511436, People's Republic of China. changlingma@gzhmu.edu.cn.
Abstract
BACKGROUND: Clonorchis sinensis was a food-borne zoonotic parasite in the worldwide and also an important risk factor of hepatic fibrosis. Excretory/secretion products of C. sinensis (CsESPs) are involved in parasite-host interactions and contribute to the development of hepatic damage. The aim of the present study was to investigate whether CsESPs and CsTP (adult protein) could induce autophagy of hepatic stellate cells (HSCs) and further activate HSCs so as to participate in the pathogenesis of hepatic fibrosis. METHODS AND RESULTS: The human hepatic stellate cell line LX-2 was stimulated by CsESPs and CsTP. CsESPs showed the effect on cell proliferation in methyl thiazolyl tetrazolium (MTT) assay while CsTP failed. Autophagosomes and autolysosomes were observed after the transmission mRFP-EGFP-LC3 plasmid into the LX-2 cells. CsESPs had more powerful to induce the accumulation of autophagosomes and autolysosomes to enhance autophagic flux compared with CsTP. Western-blotting analysis confirmed that the ratio of LC3-II/I in LX-2 cells was up-regulated after CsESPs treatment for 6 h, which further proved that CsESPs could induce autophagy in LX-2 cells. Meanwhile, q-PCR results showed that the mRNA levels of collagen I, collagen III and α-SMA decreased in LX-2 cells after treatment with autophagy inhibitor chloroquine, whereas they increased when combination with CsESPs. CONCLUSIONS: These results suggested that CsESPs-induced autophagy might be involved in the activation of HSCs, and consequently participate in the pathogenesis of hepatic fibrosis caused by C. sinensis infection.
BACKGROUND: Clonorchis sinensis was a food-borne zoonotic parasite in the worldwide and also an important risk factor of hepatic fibrosis. Excretory/secretion products of C. sinensis (CsESPs) are involved in parasite-host interactions and contribute to the development of hepatic damage. The aim of the present study was to investigate whether CsESPs and CsTP (adult protein) could induce autophagy of hepatic stellate cells (HSCs) and further activate HSCs so as to participate in the pathogenesis of hepatic fibrosis. METHODS AND RESULTS: The human hepatic stellate cell line LX-2 was stimulated by CsESPs and CsTP. CsESPs showed the effect on cell proliferation in methyl thiazolyl tetrazolium (MTT) assay while CsTP failed. Autophagosomes and autolysosomes were observed after the transmission mRFP-EGFP-LC3 plasmid into the LX-2 cells. CsESPs had more powerful to induce the accumulation of autophagosomes and autolysosomes to enhance autophagic flux compared with CsTP. Western-blotting analysis confirmed that the ratio of LC3-II/I in LX-2 cells was up-regulated after CsESPs treatment for 6 h, which further proved that CsESPs could induce autophagy in LX-2 cells. Meanwhile, q-PCR results showed that the mRNA levels of collagen I, collagen III and α-SMA decreased in LX-2 cells after treatment with autophagy inhibitor chloroquine, whereas they increased when combination with CsESPs. CONCLUSIONS: These results suggested that CsESPs-induced autophagy might be involved in the activation of HSCs, and consequently participate in the pathogenesis of hepatic fibrosis caused by C. sinensis infection.
Authors: Lien F R Thoen; Eduardo L M Guimarães; Laurent Dollé; Inge Mannaerts; Mustapha Najimi; Etienne Sokal; Leo A van Grunsven Journal: J Hepatol Date: 2011-07-29 Impact factor: 25.083
Authors: Virginia Hernández-Gea; Zahra Ghiassi-Nejad; Raphael Rozenfeld; Ronald Gordon; Maria Isabel Fiel; Zhenyu Yue; Mark J Czaja; Scott L Friedman Journal: Gastroenterology Date: 2012-01-10 Impact factor: 22.682