Effects of CpG Oligodeoxynucleotide 1826 on transforming growth factor-beta 1 and radiation-induced pulmonary fibrosis in mice
Tóm tắt
Cytosine-phosphate-guanine (CpG) oligodeoxyribonucleotides (ODNs) are synthetic DNA fragments containing unmethylated cytosine-guanine motifs with potential immune modulatory effects and have recently been suggested to enhance sensitivity to traditional therapies in lung cancer. This study aimed to examine the effects of CpG ODN1826 on transforming growth factor-beta 1(TGF-β1) and radiation-induced pulmonary fibrosis in mice. The radiation-induced pulmonary fibrosis mouse model was established by a single dose of 20 Gy, 6 MV X-rays exposure to the left lung. ICR mice were evenly randomized into four groups, comprising: a control group, a radiation group (RT group), a CpG group and a radiation combined with CpG ODN1826 group (RT + CpG group), with 40 mice in each group. CpG ODN1826 was intraperitoneally injected into mice at 1, 3, 5, 7 and 9 d post-irradiation. The mice were sacrificed at 1, 5, 15, 30 and 90 d post-irradiation. Paraffin sections of the radiated lung were subjected to H&E staining and Masson staining. The Ashcroft scale was used for quantitative histological analysis of fibrotic changes induced by irradiation. Concentrations of serum TGF-β1 were determined by ELISA, and concentrations of Hydroxyproline(Hyp) in the lung were determined with the alkaline hydrolysis method. Relative gene expression of FoxP3 was determined by real-time PCR. The radiation-induced pulmonary fibrosis mouse model was successfully established. The serum concentrations of TGF -β1 of RT group were higher than those of the RT + CpG group (t = 5.212, 7.126, 7.972 and 3.785, P < 0.05). The Hyp in the lung of RT group was higher than that of RT + CpG group (t = 4.606, P < 0.05). The relative expressions of FoxP3 gene in the lung of the RT group were higher than those of RT + CpG group (t = 8.395, 5.099 and 6.147, P < 0.05). CpG ODN1826 could reduce the serum concentrations of TGF-β1 and the lung content of Hyp in radiation-induced pulmonary fibrosis, which might be related to the possibility that CpG ODN1826 can reduce expression of the FoxP3 gene.
Tài liệu tham khảo
Jenkins P, Watts J. An improved model for predicting radiation pneumonitisincorporating clinical and dosimetric variables. Int J Radiat Oncol Biol Phys. 2011;80(4):1023–9.
Wennberg B, Gagliardi G, Sundbom L, et al. Early response of lung in breast cancer irradiation: radiologic density changes measured by CT and symptomatic radiation pneumonitis. Int J Radiat Oncol Biol Phys. 2002;52(5):1196–206.
Strieter RM. What differentiates normal lung repair and fibrosis? Inflammation, resolution of repair, and fibrosis. Proc Am Thorac Soc. 2008;5(3):305–10.
Hanagata N. Structure-dependent immunostimulatory effect of CpG oligodeoxynucleotides and their delivery system. Int J Nanomedicine. 2012;7:2181–95.
Wang YY, He XY, Cai YY, et al. The variation of CD4 + CD25+ regulatory T cells in the periphery blood and tumor microenvironment of non - small cell lung cancer patients and the downregulation effects induced by CpG ODN. Target Oncol. 2011;6(3):147–54.
Sohn BH, Park IY, Lee JJ, et al. Functional switching of TGF-beta1 signaling in liver cancer via epigenetic modulation of a single CpG site in TTP promoter. Gastroenterology. 2010;138(5):1898–908.
Arrieta O, Gallardo-Rincon D, Villarreal-Garza C, et al. High frequency of radiation pneumonitis in patients with locally advanced non-small cell lung cancer treated with concurrent radiotherapy and gemcitabine after induction with gemcitabine and carboplatin. J Thorac Oncol. 2009;4(7):845–52.
Kisseleva T, Brenner DA. Fibrogenesis of parenchymal organs. Proc Am Thorac Soc. 2008;5(3):338–42.
Kehrer JM, Lee YC, Solem SM. Comparison of in vitro and in vivo rates of collagen synthesis in normal and damaged lung tissue. Exp Lung Res. 1986;10(2):187–201.
Miranda-Hernandez DF, Franco-Molina MA, Mendoza-Gamboa E, et al. Expression of Foxp3, CD25 and IL-2 in the B16F10 cancer cell line and melanoma is correlated with tumor growth in mice. Oncol Lett. 2013;6(5):1195–200.
Peng X, Moore MW, Peng H, et al. CD4 + CD25 + FoxP3+ Tregs attenuate TGF-β1 induced lung fibrosis and fibrocyte accumulation in part via suppression of FGF-9. Front Pharmacol. 2014;5(5):80.
Sumitomo S, Fujio K, Okamura T, et al. Transcription factor early growth response 3 is associated with the TGF-β1 expression and the regulatory activity of CD4-positive T cells in vivo. J Immunol. 2013;191(5):2351–9.
Hartmann G, Krieg AM. Mechanism and function of a newly identified CpG DNA motif in human primary B cells. J Immunol. 2000;164(2):944–53.
Chace JH, Hooker NA, Mildenstein KL, et al. Bacterial DNA-induced NK cell IFN-gamma production is dependent on macrophage secretion of IL-12. Clin Immunol Immunopathol. 1997;84(2):185–93.
Cooper CL, Davis HL, Morris ML, et al. CPG 7909, an immunostimulatory TLR9 agonist oligodeoxynucleotide, as adjuvant to Engerix-B HBV vaccine in healthy adults: a double-blind phase I/II study. J Clin Immunol. 2004;24(6):693–701.
Cooper CL, Davis HL, Morris ML, et al. Safety and immunogenicity of CPG 7909 injection as an adjuvant to Fluarix influenza vaccine. Vaccine. 2004;22(23–24):3136–43.
Krieg AM. CpG still rocks! Update on an accidental drug. Nucleic Acid Ther. 2012;22(2):77–89.
Wright SE, Rewers-Felkins KA, Chowdhury NI, et al. Prevention of human adenocarcinoma with CpG-ODN in a mouse model. Oncol Lett. 2012;4(5):1061–3.
Mueller M, Reichardt W, Koerner J, et al. Coencapsulation of tumor lysate and CpG-ODN in PLGA-microspheres enables successful immunotherapy of prostate carcinoma in TRAMP mice. J Control Release. 2012;162(1):159–66.
Pali-Scholl I, Szollosi H, Starkl P, et al. Protamine nanoparticles with CpG-oligodeoxynucleotide prevent an allergen-induced Th2-response in BALB/c mice. Eur J Pharm Biopharm. 2013;85(3):656–64.
Zha L, Qiao T, Yuan S, et al. Enhancement of radiosensitivity by CpG-oligodeoxyribonucleotide-7909 in human non-small cell lung cancer A549 cells. Cancer Biother Radiopharm. 2010;25(2):165–70.
Wilson MS, Wynn TA. Pulmonary fibrosis: pathogenesis, etiology and regulation. Mucosal Immunol. 2009;2(2):103–21.
Szabo S, Ghosh SN, Fish BL, et al. Cellular inflammatory infiltrate in pneumonitis induced by a single moderate dose of thoracic x radiation in rats. Radiat Res. 2010;173(4):545–56.
Kiazimov KI. Pathogenesis, diagnostics and treatment of radiation pneumonitis induced by radiotherapy of lung cancer. Georgian Med News. 2009;174:115–8.
Rube CE, Uthe D, Schmid KW, et al. Dose - dependent induction of transforming growth factor beta (TGF-beta) in the lung tissue of fibrosis - pronemice after thoracic irradiation. Int J Radiat Oncol Biol Phys. 2000;47(4):1033–42.
Willis BC, Liebler JM, Luby-Phelps K, et al. Induction of epithelial - mesenchymal transition in alveolar epithelial cells by transforming growth factor - beta 1: potential role in idiopathic pulmonary fibrosis. Am J Pathol. 2005;166(5):1321–32.
Wills BC, Borok Z. TGF – beta - induced EMT: mechanisms and implications for fibrotic lung disease. Am J Physiol Lung Cell Mol Physiol. 2007;293(3):525–34.
Li H, Yu JP, Cao S, et al. CD4 + CD25 + regulatory T cells decreased the antitumor activity of cytokine - induced killer (CIK) cells of lung cancer patients. J Clin Immunol. 2007;27(3):317–26.
Gallimore A, Godkin A. Regulatory T cells and tumour immunity - observations in mice and men. Immunology. 2008;123(2):157–63.
Molteni A, Wolfe LF, Ward WF, et al. Effect of an angiotensin II receptor blocker and two angiotensin converting enzyme inhibitors ontransforming growth factor-beta (TGF-beta) and alpha – actomyosin (alpha SMA), important mediators of radiation -induced pneumopathy and lung fibrosis. Curr Pharm Des. 2007;13(13):1307–16. 31.
Jain R, Shaul PW, Borok Z, et al. Endothelin-1 induces alveolar EMT through ET-A-mediated production of transforming growth factor-β1. Am J Respir Cell Mol Biol. 2007;37(1):38–47.