The roles of Toll-like receptor 4 in the pathogenesis of pathogen-associated biliary fibrosis caused by Clonorchis sinensis

Yan, Chao; Li, Bo; Fan, Fang; Du, Ying; Ma, Rui; Cheng, Xiao-Dan; Li, Xiangyang; Zhang, Bo; Yu, Qian; Wang, Yugang; Tang, Renxian; Zheng, Kuiyang

doi:10.1038/s41598-017-04018-8

Cited by 25 publications

(25 citation statements)

References 37 publications

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“…In addition, in the presence of ESP and NDMA stimulation, Cx43 knockdown inhibited Cox-2 expression in H69 cells. Yan et al [15] reported that iNOS is highly expressed in Kupffer cells, sinusoidal endothelial cells, and biliary epithelial cells in BALB/c mice after C. sinensis infection. Nitric oxide (NO) formation and nitrosation may contribute to the development of C. sinensis -associated carcinogenesis [11, 12, 14, 15].…”

Section: Discussionmentioning

confidence: 99%

“…Yan et al [15] reported that iNOS is highly expressed in Kupffer cells, sinusoidal endothelial cells, and biliary epithelial cells in BALB/c mice after C. sinensis infection. Nitric oxide (NO) formation and nitrosation may contribute to the development of C. sinensis -associated carcinogenesis [11, 12, 14, 15]. The induction of iNOS under inflammatory conditions suggests that NO is involved in the upregulation of Cx43 [16, 19].…”

Section: Discussionmentioning

confidence: 99%

“…CHCA is potentially caused by increased levels of proinflammatory cytokines and nuclear factor kappa B (NFκB), which regulate the activities of cyclooxygenase-2 (Cox-2) and inducible nitric oxide synthase, and disturb the homeostasis of oxidants/anti-oxidants and DNA repair enzymes [13]. ESPs of C. sinensis induce pro-inflammatory responses in vitro by the upregulation of TLR4 and its downstream transduction signals, including MyD88-dependent IκB-α degradation and NFκB activation [14, 15]. NFκB may also influence the production of Cx43, a gap-junction protein, in liver cirrhosis [16, 17].…”

Section: Introductionmentioning

confidence: 99%

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Connexin 43 plays an important role in the transformation of human cholangiocytes upon stimulation withClonochis sinensisexcretory-secretory protein andN-nitrosodimethylamine

Kim

Bae

Choi³

et al. 2018

Preprint

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BackgroundClonorchis sinensis is a group I bio-carcinogen responsible for cholangiocarcinoma (CHCA) in humans. However, the mechanism by which C. sinensis promotes carcinogenesis is unclear.MethodologyUsing the human cholangiocyte line H69, we investigated cell proliferation and gap junction protein expression after stimulation with the hepatotoxin N-nitrosodimethylamine (NDMA) and/or excretory-secretory products (ESP) of C. sinensis, which induce inflammation. NDMA and ESP treatment increased proliferation by 146% and the proportion of cells in the G2/M phase by 37%. Moreover, the expression of the cell cycle protein E2F1 and the cell proliferation-related proteins Ki-67 and cytokeratin 19 increased in response to combined treatment with NDMA and ESP. The gap-junction proteins connexin (Cx) 43 and Cx26 also increased. In contrast, Cx32 expression decreased in cells treated with NDMA and ESP. Cox-2 was also upregulated. Silencing of Cx43 reduced cell proliferation and significantly suppressed Cx26 and Cox-2 expression.ConclusionsThese results suggest that Cx43 is an important factor in CHCA induced by C. sinensis ESP and NDMA and further investigations targeting this pathway may allow prevention of this deadly disease.Author summaryClonorchis sinensis, a human fluke, resides in the liver of humans and is commonly found in the common bile duct and gall bladder. This parasite is the main cause of cholangiocarcinoma, also called bile duct cancer, in humans. Of note, the excretory-secretory products (ESP) of C. sinensis are known to cause inflammation in the biliary epithelium, which may ultimately result in neoplasms via production of reactive oxygen species and subsequent DNA damage. Together with N-nitrosodimethylamine (NDMA), a potent hepatotoxin that can cause fibrosis and tumors in the liver, ESP led to an increase in the growth and proliferation of cholangiocytes. Our results showed that the ESPs of C. sinensis induced pro-inflammatory responses by increasing the levels of proinflammatory cytokines and nuclear factor kappa B (NFκB), which in turn, enhanced the production of connexin 43 (Cx43), a gap-junction protein. Therefore, Cx 43 can serve as a potential target for developing a therapeutic strategy for the treatment of cholangiocarcinoma in humans.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Connexin 43 plays an important role in the transformation of human cholangiocytes upon stimulation withClonochis sinensisexcretory-secretory protein andN-nitrosodimethylamine

Kim

Bae

Choi³

et al. 2018

Preprint

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“…The concentration of hepatic hydroxyproline was determined as described in our recent publication 30 . Briefly, frozen liver tissue was heated in 6 mM HCl at 120 °C for 4 h. After centrifugation, the supernatant was mixed with chloramine-T solution (1.4% chloramine-T, 10% N-propanol, and 80% citrate-acetate buffer) for 20 min at room temperature and followed by adding Ehrlich’s solution at 65 °C for 20 min.…”

Section: Methodsmentioning

confidence: 99%

The gut microbiota regulates mouse biliary regenerative responses

Liu¹,

Yan²,

Zhang³

et al. 2019

Preprint

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17Injury to the biliary epithelium triggers cholangiopathies. However, factors involved in 18 regulating the pathogenesis remains incompletely understood. Here, we report that the 19 gut microbiota is important in regulating hepatobiliary fibroproliferative regenerative 20 process. In helminth Clonorchis sinensis-induced bile duct injury model, wild-type mice 21 showed more extensive peribiliary fibroproliferative responses than non-littermate IL-33-22 deficient mice. However, these reactions could be attenuated by co-housing of the 23 animals together. In the meantime, the relatively fibroproliferative-resistant IL-33-24 deficient mice could become fibroproliferative-responsive especially in large intrahepatic 25 bile duct by antibiotics treatment. Furthermore, microbiota-derived metabolite butyrate 26 was able to inhibit biliary organoid expansion in vitro and temper 3,5-diethoxycarbonyl-27 1,4-dihydrocollidine-induced biliary fibrosis in vivo. Together, our data implies a potential 28 way of management of hepatobiliary diseases by modulating gut microbiota. 29 30 31 3 32 33The biliary tree including intrahepatic and extrahepatic bile ducts (IHBD and EHBD) 34 forms the primary path by which bile is secreted by the liver then transported to the gut. 35Disruption of the epithelial lining of bile ducts triggers hepatobiliary inflammation, 36 fibrosis, and even malignant transformation, which collectively were referred as 37 cholangiopathies. The cholangiopathies account for substantial morbidity and mortality 38 and are often progressive that lead to end-stage liver disease. Despite the severe 39 disease phenotypes, the molecular and cellular effectors and their relationships to 40 cholangiopathies remain poorly defined 1 . 41A number of animal models have been generated to provide in vivo tools to 42 investigate the mechanisms of biliary repair and, eventually, to test the efficacy of 43 innovative treatments 2 . Each of the animal model has its own limitations in translating 44 to human settings. 3,5-Diethoxycarbonyl-1,4-dihydrocollidine (DDC)-feeding in mice 45 recapitulated pathogenic hallmarks of human primary sclerosing cholangitis (PSC) 3 . 46Clonorchis sinensis is a food-borne helminth that naturally cause cholangiopathies 4,5 . 47During progression of biliary injury induced by the worm, a disorganized expansion of 48 bile ducts and recruitment of inflammatory cells known as ductular reaction (DR) 49 become so prominent that not only biliary epithelial but also the peribiliary glands 50 (PBGs), clusters of epithelial cells residing in the submucosal compartment of the bile 51 duct, show hyperplasia and proliferation. PBGs participate in the secretion of neutral, 52 carboxylated, and sulphated mucins into the bile duct lumen. The term adenomatous 53 hyperplasia or chronic proliferative cholangitis has been used to describe this 54 4 phenomenon 6 . What is the underlying molecular and cellular mechanisms of these 55 biliary responses remains incompletely understood. 56The gut microbiota can influence or m...

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“…Thirty mice randomly divided into 10 mice per group were used to prepare serum samples of C. sinensis, S. japonicum and P. berghei infection. Each mouse was gavaged with 45 metacercariae, and C. sinensispositive serum was collected one month later [22]. S. japonicum-positive serum was prepared by infecting mice (40 cercariae per mouse) percutaneously in a shaved region of the abdomen [23].…”

Section: Preparation Of Rtgprxmentioning

confidence: 99%

Evaluation of toxoplasmosis in pregnant women using dot-immunogold-silver staining with recombinant Toxoplasma gondii peroxiredoxin protein

Liu

et al. 2020

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Background: Toxoplasma gondii infection endangers human health and affects animal husbandry. Serological detection is the main method used for epidemiological investigations and diagnosis of toxoplasmosis. The key to effective diagnosis of toxoplasmosis is the use of a standardized antigen and a specific and sensitive detection method. Peroxiredoxin is an antigenic protein and vaccine candidate antigen of T. gondii that has not yet been exploited for diagnostic application. Methods: In this study, recombinant T. gondii peroxiredoxin protein (rTgPrx) was prepared and used in dot-immunogold-silver staining (Dot-IGSS) to detect IgG antibodies in serum from mice and pregnant women. The rTgPrx-Dot-IGSS method was established and optimized using mouse serum. Furthermore, serum samples from pregnant women were analyzed by rTgPrx-Dot-IGSS.Results: Forty serum samples from mice infected with T. gondii and twenty negative serum samples were analyzed. The sensitivity and specificity of rTgPrx-Dot-IGSS were 97.5% and 100%, respectively, equivalent to those of a commercial ELISA kit for anti-Toxoplasma IgG antibody. Furthermore, 540 serum samples from pregnant women were screened with a commercial ELISA kit. Eighty-three positive and 60 negative serum samples were analyzed by rTgPrx-Dot-IGSS. The positive rate was 95.18%, comparable to that obtained with the commercial ELISA kit.Conclusions: The Dot-IGSS method with rTgPrx as an antigen might be useful for diagnosing T. gondii infection in individuals.

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The roles of Toll-like receptor 4 in the pathogenesis of pathogen-associated biliary fibrosis caused by Clonorchis sinensis

Cited by 25 publications

References 37 publications

Connexin 43 plays an important role in the transformation of human cholangiocytes upon stimulation withClonochis sinensisexcretory-secretory protein andN-nitrosodimethylamine

Connexin 43 plays an important role in the transformation of human cholangiocytes upon stimulation withClonochis sinensisexcretory-secretory protein andN-nitrosodimethylamine

The gut microbiota regulates mouse biliary regenerative responses

Evaluation of toxoplasmosis in pregnant women using dot-immunogold-silver staining with recombinant Toxoplasma gondii peroxiredoxin protein

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