Connective tissue growth factor stimulates the proliferation, migration and differentiation of lung fibroblasts during paraquat‑induced pulmonary fibrosis

Yang,Zhizhou; Sun,Zhaorui; Liu,Hongmei; Ren,Yi; Shao,Danbing; Zhang,Wei; Lin,Jinfeng; Wolfram,Joy; Wang,Feng; Nie,Shinan

doi:10.3892/mmr.2015.3537

Connective tissue growth factor stimulates the proliferation, migration and differentiation of lung fibroblasts during paraquat‑induced pulmonary fibrosis

Authors:
- Zhizhou Yang
- Zhaorui Sun
- Hongmei Liu
- Yi Ren
- Danbing Shao
- Wei Zhang
- Jinfeng Lin
- Joy Wolfram
- Feng Wang
- Shinan Nie
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Affiliations: Department of Emergency Medicine, Jinling Hospital, Medical School of Nanjing University, Nanjing, Jiangsu 210002, P.R. China, CAS Key Laboratory for Biomedial Effects of Nanomaterials & Nanosafety, National Center for Nanoscience and Technology of China, University of Chinese Academy of Sciences, Beijing 100190, P.R. China, Department of Gastroenterology, The Tenth People's Hospital of Shanghai, Tongji University, Shanghai 200072, P.R. China
Published online on: March 24, 2015 https://doi.org/10.3892/mmr.2015.3537
Pages: 1091-1097
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

It is well established that paraquat (PQ) poisoning can cause severe lung injury during the early stages of exposure, finally leading to irreversible pulmonary fibrosis. Connective tissue growth factor (CTGF) is an essential growth factor that is involved in tissue repair and pulmonary fibrogenesis. In the present study, the role of CTGF was examined in a rat model of pulmonary fibrosis induced by PQ poisoning. Histological examination revealed interstitial edema and extensive cellular thickening of interalveolar septa at the early stages of poisoning. At 2 weeks after PQ administration, lung tissue sections exhibited a marked thickening of the alveolar walls with an accumulation of interstitial cells with a fibroblastic appearance. Masson's trichrome staining revealed a patchy distribution of collagen deposition, indicating pulmonary fibrogenesis. Western blot analysis and immunohistochemical staining of tissue samples demonstrated that CTGF expression was significantly upregulated in the PQ‑treated group. Similarly, PQ treatment of MRC‑5 human lung fibroblast cells caused an increase in CTGF in a dose‑dependent manner. Furthermore, the addition of CTGF to MRC‑5 cells triggered cellular proliferation and migration. In addition, CTGF induced the differentiation of fibroblasts to myofibroblasts, as was evident from increased expression of α‑smooth muscle actin (α‑SMA) and collagen. These findings demonstrate that PQ causes increased CTGF expression, which triggers proliferation, migration and differentiation of lung fibroblasts. Therefore, CTGF may be important in PQ‑induced pulmonary fibrogenesis, rendering this growth factor a potential pharmacological target for reducing lung injury.

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1	Dinis-Oliveira RJ, Duarte JA, Sánchez-Navarro A, et al: Paraquat poisonings: mechanisms of lung toxicity, clinical features, and treatment. Crit Rev Toxicol. 38:13–71. 2008. View Article : Google Scholar
2	Neves FF, Sousa RB, Pazin-Filho A, et al: Severe paraquat poisoning: clinical and radiological findings in a survivor. J Bras Pneumol. 36:513–516. 2010. View Article : Google Scholar : PubMed/NCBI
3	Chen CM and Lua AC: Lung toxicity of paraquat in the rat. J Toxicol Environ Health A. 60:477–487. 2000. View Article : Google Scholar
4	Baltazar T, Dinis-Oliveira RJ, Duarte JA, et al: Paraquat research: do recent advances in limiting its toxicity make its use safer? Br J Pharmacol. 168:44–45. 2013. View Article : Google Scholar :
5	Novaes RD, Gonçalves RV, Cupertino MC, et al: Bark extract of Bathysa cuspidata attenuates extra-pulmonary acute lung injury induced by paraquat and reduces mortality in rats. Int J Exp Pathol. 93:225–233. 2012. View Article : Google Scholar : PubMed/NCBI
6	Li LR, Sydenham E, Chaudhary B and You C: Glucocorticoid with cyclophosphamide for paraquat-induced lung fibrosis. Cochrane Database Syst Rev. 16:CD0080842010.
7	Lin JL, Lin-Tan DT, Chen KH, et al: Improved survival in severe paraquat poisoning with repeated pulse therapy of cyclophosphamide and steroids. Intensive Care Med. 37:1006–1013. 2011. View Article : Google Scholar : PubMed/NCBI
8	Moon JM and Chun BJ: The efficacy of high doses of vitamin C in patients with paraquat poisoning. Hum Exp Toxicol. 30:844–850. 2011. View Article : Google Scholar
9	Suntres ZE: Role of antioxidants in paraquat toxicity. Toxicology. 180:65–77. 2002. View Article : Google Scholar : PubMed/NCBI
10	Koo JR, Kim JC, Yoon JW, et al: Failure of continuous venovenous hemofiltration to prevent death in paraquat poisoning. Am J Kidney Dis. 39:55–59. 2002. View Article : Google Scholar : PubMed/NCBI
11	Koo JR, Yoon JW, Han SJ, et al: Rapid analysis of plasma paraquat using sodium dithionite as a predictor of outcome in acute paraquat poisoning. Am J Med Sci. 338:373–377. 2009. View Article : Google Scholar : PubMed/NCBI
12	Harlow CR and Hillier SG: Connective tissue growth factor in the ovarian paracrine system. Mol Cell Endocrinol. 187:23–27. 2002. View Article : Google Scholar : PubMed/NCBI
13	Takigawa M, Nakanishi T, Kubota S and Nishida T: Role of CTGF/HCS24/ecogenin in skeletal growth control. J Cell Physiol. 194:256–266. 2003. View Article : Google Scholar : PubMed/NCBI
14	Wang Q, Usinger W, Nichols B, et al: Cooperative interaction of CTGF and TGF-β in animal models of fibrotic disease. Fibrogenesis Tissue Repair. 4:42011. View Article : Google Scholar
15	Moussad EE and Brigstock DR: Connective tissue growth factor: what’s in a name? Mol Genet Metab. 71:276–292. 2000. View Article : Google Scholar : PubMed/NCBI
16	Ponticos M, Holmes AM, Shi-wen X, et al: Pivotal role of connective tissue growth factor in lung fibrosis: MAPK-dependent transcriptional activation of type I collagen. Arthritis Rheum. 60:2142–2155. 2009. View Article : Google Scholar : PubMed/NCBI
17	Bogatkevich GS, Ludwicka-Bradley A, Singleton CB, et al: Proteomic analysis of CTGF-activated lung fibroblasts: identification of IQGAP1 as a key player in lung fibroblast migration. Am J Physiol Lung Cell Mol Physiol. 295:L603–L611. 2008. View Article : Google Scholar : PubMed/NCBI
18	Shimo T, Kubota S, Kondo S, et al: Connective tissue growth factor as a major angiogenic agent that is induced by hypoxia in a human breast cancer cell line. Cancer Lett. 174:57–64. 2001. View Article : Google Scholar : PubMed/NCBI
19	Chujo S, Shirasaki F, Kawara S, et al: Connective tissue growth factor causes persistent proalpha2(I) collagen gene expression induced by transforming growth factor-beta in a mouse fibrosis model. J Cell Physiol. 203:447–456. 2005. View Article : Google Scholar
20	Heng EC, Huang Y, Black SA Jr and Trackman PC: CCN2, connective tissue growth factor, stimulates collagen deposition by gingival fibroblasts via module 3 and alpha6- and beta1 integrins. J Cell Biochem. 98:409–420. 2006. View Article : Google Scholar : PubMed/NCBI
21	Sun Z, Gong X, Zhu H, et al: Inhibition of Wnt/β-catenin signaling promotes engraftment of mesenchymal stem cells to repair lung injury. J Cell Physiol. 229:213–224. 2014. View Article : Google Scholar
22	McAnulty RJ: Fibroblasts and myofibroblasts: their source, function and role in disease. Int J Biochem Cell Biol. 39:666–671. 2007. View Article : Google Scholar : PubMed/NCBI
23	Scotton CJ and Chambers RC: Molecular targets in pulmonary fibrosis: the myofibroblast in focus. Chest. 132:1311–1321. 2007. View Article : Google Scholar : PubMed/NCBI
24	Bonniaud P, Margetts PJ, Kolb M, et al: Adenoviral gene transfer of connective tissue growth factor in the lung induces transient fibrosis. Am J Respir Crit Care Med. 168:770–778. 2003. View Article : Google Scholar : PubMed/NCBI
25	Chen CM, Wang LF, Chou HC, Lang YD and Lai YP: Up-regulation of connective tissue growth factor in hyperoxia-induced lung fibrosis. Pediatr Res. 62:128–133. 2007. View Article : Google Scholar : PubMed/NCBI
26	Sun Z, Wang C, Shi C, et al: Activated Wnt signaling induces myofibroblast differentiation of mesenchymal stem cells, contributing to pulmonary fibrosis. Int J Mol Med. 33:1097–1109. 2014.PubMed/NCBI
27	Phan SH: The myofibroblast in pulmonary fibrosis. Chest. 122(6 Suppl): 286S–289S. 2002. View Article : Google Scholar : PubMed/NCBI
28	Ahmed AA: Protective effect of montelukast on paraquat-induced lung toxicity in rats. Biosci Trends. 3:63–72. 2009.
29	Tomita M, Okuyama T, Katsuyama H, et al: Mouse model of paraquat-poisoned lungs and its gene expression profile. Toxicology. 231:200–209. 2007. View Article : Google Scholar : PubMed/NCBI
30	Hoet PH and Nemery B: Polyamines in the lung: polyamine uptake and polyamine-linked pathological or toxicological conditions. Am J Physiol Lung Cell Mol Physiol. 278:L417–L433. 2000.PubMed/NCBI
31	Smith LL: Mechanism of paraquat toxicity in lung and its relevance to treatment. Hum Toxicol. 6:31–36. 1987. View Article : Google Scholar : PubMed/NCBI
32	Smith LL, Lewis CP, Wyatt I and Cohen GM: The importance of epithelial uptake systems in lung toxicity. Environ Health Perspect. 85:25–30. 1990. View Article : Google Scholar : PubMed/NCBI
33	Lacerda AC, Rodrigues-Machado Mda G, Mendes PL, et al: Paraquat (PQ)-induced pulmonary fibrosis increases exercise metabolic cost, reducing aerobic performance in rats. J Toxicol Sci. 34:671–679. 2009. View Article : Google Scholar : PubMed/NCBI
34	Tung JN, Lang YD, Wang LF and Chen CM: Paraquat increases connective tissue growth factor and collagen expression via angiotensin signaling pathway in human lung fibroblasts. Toxicol In Vitro. 24:803–808. 2010. View Article : Google Scholar
35	Bhogal RK, Stoica CM, McGaha TL and Bona CA: Molecular aspects of regulation of collagen gene expression in fibrosis. J Clin Immunol. 25:592–603. 2005. View Article : Google Scholar : PubMed/NCBI
36	Phan SH: Biology of fibroblasts and myofibroblasts. Proc Am Thorac Soc. 5:334–337. 2008. View Article : Google Scholar : PubMed/NCBI
37	Kendall RT and Feghali-Bostwick CA: Fibroblasts in fibrosis: novel roles and mediators. Front Pharmacol. 5:1232014. View Article : Google Scholar : PubMed/NCBI
38	Liu X, Wu W and Chen H: Effects of CTGF gene silencing on the proliferation and myofibroblast differentiation of human lung fibroblasts. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 25:407–412. 2008.In Chinese. PubMed/NCBI