The expression dynamics of transforming growth factor-β/Smad signaling in the liver fibrosis experimentally caused by Clonorchis sinensis

Particle and Fibre Toxicology

Kui-Yang Zheng¹  Ren-Xian Tang¹  Jia-Xu Chen²  Xiang-Yang Li¹  Yan-Hong Wang¹  Bo Zhang¹  Bei-Bei Zhang¹  Bo Li¹  Lin Wang¹  Chao Yan¹ 

[1] Department of Pathogenic Biology and Immunology, Laboratory of Infection and Immunity, Xuzhou Medical College, Xuzhou 221004, Jiangsu Province, PR China;Laboratory of Parasite and Vector Biology, National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Shanghai 200025, PR China

关键词: Smads;    Transforming growth factor-β;    Liver fibrosis;    Clonorchis sinensis;   

Others  :  1147203
DOI  :  10.1186/s13071-015-0675-y

 received in 2014-12-16, accepted in 2015-01-20,  发布年份 2015

【 摘 要 】

Background

Liver fibrosis is a hallmark of clonorchiasis suffered by millions people in Eastern Asian countries. Recent studies showed that the activation of TGF-β/Smad signaling pathway can potently regulate the hepatic fibrogenesis including Schistosoma spp. and Echinococcus multilocularis-caused liver fibrosis. However, little is known to date about the expression of transforming growth factor-β (TGF-β) and other molecules in TGF-β/Smad signaling pathway which may play an important role in hepatic fibrosis caused by C. sinensis.

Methods

A total of 24 mice were individually infected orally with 45 metacercariae, both experimental mice and mocked-infected control mice were anesthetized at 4 week post-infection (wk p.i.), 8 wk p.i. and 16 wk p.i., respectively. For each time-point, the liver and serum from each animal were collected to analyze histological findings and various fibrotic parameters including TGF-β₁, TGF-β receptors and down-stream Smads activation, as well as fibrosis markers expression.

Results

The results showed that collagen deposition indicated by hydroxyproline content and Masson’s trichrome staining was increased gradually with the development of infection. The expression of collagen type α1 (Col1a) mRNA transcripts was steadily increased during the whole infection. The mRNA levels of Smad2, Smad3 as well as the protein of Smad3 in the liver of C. sinensis-infected mice were increased after 4 wk p.i. (P < 0.05, compared with normal control) whereas the TGF-β₁, TGF-β type I receptor (TGFβRI) and TGF-β type II receptor (TGFβRII) mRNA expression in C. sinensis-infected mice were higher than those of normal control mice after 8 wk p.i. (P < 0.05). However, the gene expression of Smad4 and Smad7 were peaked at 4 wk p.i. (P < 0.05), and thereafter dropped to the basal level at 8 wk p.i., and 16 wk p.i., respectively. The concentrations of TGF-β₁ in serum in the C. sinensis-infected mice at 8 wk p.i. and 16 wk p.i (P < 0.05) were significantly higher than those in the control mice.

Conclusions

The results of the present study indicated for the first time that the activation of TGF-β/Smad signaling pathway might contribute to the synthesis of collagen type I which leads to liver fibrosis caused by C. sinensis.

【 授权许可】

   
2015 Yan et al,; licensee BioMed Central.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Fang YY, Chen YD, Li XM, Wu J, Zhang QM, Ruan CW. Current prevalence of Clonorchis sinensis infection in endemic areas of China. ZhongguoJi Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 2008, 26:99–103, 109.
• [2]Hong ST, Fang Y. Clonorchis sinensis and clonorchiasis, an update. Parasitol Int. 2012; 61:17-24.
• [3]Li S, Chung YB, Chung BS, Choi MH, Yu JR, Hong ST. The involvement of the cysteine proteases of Clonorchis sinensis metacercariae in excystment. Parasitol Res. 2004; 93:36-40.
• [4]Kim TI, Yoo WG, Kwak BK, Seok JW, Hong SJ. Tracing of the Bile-chemotactic migration of juvenile Clonorchis sinensis in rabbits by PET-CT. PLoS Negl Trop Dis. 2011; 5:e1414.
• [5]Chen J, Xu MJ, Zhou DH, Song HQ, Wang CR, Zhu XQ. Canine and feline parasitic zoonoses in China. Parasit Vectors. 2012; 5:152. BioMed Central Full Text
• [6]Bouvard V, Baan R, Straif K, Grosse Y, Secretan B, El Ghissassi F et al.. A review of human carcinogens–Part B: biological agents. Lancet Oncol. 2009; 10:321-2.
• [7]Yoon BI, Choi YK, Kim DY, Hyun BH, Joo KH, Rim HJ et al.. Infectivity and pathological changes in murine clonorchiasis: comparison in immunocompetent and immunodeficient mice. J Vet Med Sci. 2001; 63:421-5.
• [8]Choi YK, Yoon BI, Won YS, Lee CH, Hyun BH, Kim HC et al.. Cytokine responses in mice infected with Clonorchis sinensis. Parasitol Res. 2003; 91:87-93.
• [9]Wang X, Hu F, Hu X, Chen W, Huang Y, Yu X. Proteomic identification of potential Clonorchis sinensis excretory/secretory products capable of binding and activating human hepatic stellate cells. Parasitol Res. 2014; 113:3063-71.
• [10]Liang P, Sun J, Huang Y, Zhang F, Zhou J, Hu Y et al.. Biochemical characterization and functional analysis of fructose-1,6-bisphosphatase from Clonorchis sinensis. Mol Biol Rep. 2013; 40:4371-82.
• [11]Zhang F, Liang P, Chen W, Wang X, Hu Y, Liang C et al.. Stage-specific expression, immunolocalization of Clonorchissinensis lysophospholipase and its potential role in hepatic fibrosis. Parasitol Res. 2013; 112:737-49.
• [12]Zheng M, Hu K, Liu W, Hu X, Hu F, Huang L et al.. Proteomic analysis of excretory secretory products from Clonorchis sinensis adult worms: molecular characterization and serological reactivity of a excretory-secretory antigen-fructose-1,6-bisphosphatase. Parasitol Res. 2011; 109:737-44.
• [13]Hu F, Hu X, Ma C, Zhao J, Xu J, Yu X. Molecular characterization of a novel Clonorchis sinensis secretory phospholipase A(2) and investigation of its potential contribution to hepatic fibrosis. Mol Biochem Parasitol. 2009; 167:127-34.
• [14]Kanzler S, Lohse AW, Keil A, Henninger J, Dienes HP, Schirmacher P et al.. TGF-beta1 in liver fibrosis: an inducible transgenic mouse model to study liver fibrogenesis. Am J Physiol. 1999; 276:1059-68.
• [15]Dooley S, Delvoux B, Streckert M, Bonzel L, Stopa M, ten Dijke P et al.. Transforming growth factor beta signal transduction in hepatic stellate cells via Smad2/3 phosphorylation, a pathway that is abrogated during in vitro progression to myofibroblasts. TGFbeta signal transduction during transdifferentiation of hepatic stellate cells. FEBS Lett. 2001; 502:4-10.
• [16]Schiller M, Javelaud D, Mauviel A. TGF-beta-induced SMAD signaling and gene regulation: consequences for extracellular matrix remodeling and wound healing. J Dermatol Sci. 2004; 35:83-92.
• [17]Yoshida K, Murata M, Yamaguchi T, Matsuzaki K. TGF-β/Smad signaling during hepatic fibro-carcinogenesis (review). Int J Oncol. 2014; 45:1363-71.
• [18]Date M, Matsuzaki K, Matsushita M, Sakitani K, Shibano K, Okajima A et al.. Differential expression of transforming growth factor-beta and its receptors in hepatocytes and nonparenchymal cells of rat liver after CCl4 administration. J Hepatol. 1998; 28:572-81.
• [19]Yoshida K, Matsuzaki K. Differential regulation of TGF-β/Smad signaling in hepatic stellate cells between acute and chronic liver injuries. Front Physiol. 2012; 3:53.
• [20]Wang J, Zhang C, Wei X, Blagosklonov O, Lv G, Lu X et al.. TGF-β and TGF-β/Smad signaling in the interactions between Echinococcus multilocularis and its hosts. PLoS One. 2013; 8:e55379.
• [21]Zhang BB, Cai WM, Tao J, Zheng M, Liu RH. Expression of Smad proteins in the process of liver fibrosis in mice infected with Schistosoma japonicum. Zhong guo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2013; 31:89-94.
• [22]Chen BL, Peng J, Li QF, Yang M, Wang Y, Chen W. Exogenous bone morphogenetic protein-7 reduces hepatic fibrosis in Schistosomajaponicum-infected mice via transforming growth factor-β/Smad signaling. World J Gastroenterol. 2013; 19:1405-15.
• [23]Bataller R, Brenner DA. Liver fibrosis. J Clin Invest. 2005; 115:209-18.
• [24]Liu Y, Munker S, Müllenbach R, Weng HL. IL-13 signaling in liver fibrogenesis. Front Immunol. 2012; 3:116.
• [25]Chuah C, Jones MK, Burke ML, McManus DP, Gobert GN. Cellular and chemokine-mediated regulation in schistosome-induced hepatic pathology. Trends Parasitol. 2014; 30:141-50.
• [26]Tang J, Huang H, Ji X, Zhu X, Li Y, She M et al.. Involvement of IL-13 and tissue transglutaminase in liver granuloma and fibrosis after schistosoma japonicum infection. Mediators Inflamm. 2014; 2014:753483.
• [27]Tan Z, Qian X, Jiang R, Liu Q, Wang Y, Chen C et al.. IL-17A plays a critical role in the pathogenesis of liver fibrosis through hepatic stellate cell activation. J Immunol. 2013; 191:1835-44.
• [28]Chen N, Geng Q, Zheng J, He S, Huo X, Sun X. Suppression of the TGF-β/Smad signaling pathway and inhibition of hepatic stellate cell proliferation play a role in the hepatoprotective effects of curcumin against alcohol-induced hepatic fibrosis. Int J Mol Med. 2014; 34:1110-6.
• [29]Dooley S, ten Dijke P. TGF-β in progression of liver disease. Cell Tissue Res. 2012; 347:245-56.
• [30]Zhu D, He X, Duan Y, Chen J, Wang J, Sun X et al.. Expression of microRNA-454 in TGF-β1-stimulated hepatic stellate cells and in mouse livers infected with Schistosoma japonicum. Parasit Vectors. 2014; 7:148. BioMed Central Full Text
• [31]El-Lakkany NM, Hammam OA, El-Maadawy WH, Badawy AA, Ain-Shoka AA, Ebeid FA. Anti-inflammatory/anti-fibrotic effects of the hepatoprotectivesilymarin and the schistosomicide praziquantel against Schistosoma mansoni-induced liver fibrosis. Parasit Vectors. 2012; 5:9. BioMed Central Full Text
• [32]Barros AF, Oliveira SA, Carvalho CL, Silva FL, de Souza VC, da Silva AL et al.. Low transformation growth factor-β1 production and collagen synthesis correlate with the lack of hepatic periportal fibrosis development in undernourished mice infected with Schistosoma mansoni. Mem Inst Oswaldo Cruz. 2014; 109:210-9.
• [33]Attia YM, Elalkamy EF, Hammam OA, Mahmoud SS, El-Khatib AS. Telmisartan, an AT1 receptor blocker and a PPAR gamma activator, alleviates liver fibrosis induced experimentally by Schistosoma mansoni infection. Parasit Vectors. 2013; 6:199. BioMed Central Full Text
• [34]Wang HQ, Ding XD, Wu Q, Zhang Q, Huang Y, Yang F. Expression of transforming growth factor–beta and Smads in hepatic fibrosis induced by Schistosoma Japonica in mice. World J Gastroenterol. 2008; 16:929-34.
• [35]Sobral LM, Montan PF, Zecchin KG, Martelli-Junior H, Vargas PA, Graner E et al.. Smad7 blocks transforming growth factor-β1-induced gingival fibroblast-myofibroblast transition via inhibitory regulation of Smad2 and connective tissue growth factor. J Periodontol. 2011; 82:642-51.
• [36]Kamiya Y, Miyazono K, Miyazawa K. Smad7 inhibits transforming growth factor-beta family type i receptors through two distinct modes of interaction. J Biol Chem. 2010; 285:30804-13.