Anti-inflammatory and Anti-apoptotic Effect of Valproic Acid and Doxycycline Independent from MMP Inhibition in Early Radiation Damage

Yıl 2016, Cilt: 33 Sayı: 5, 488 - 495, 01.09.2016

Ferda Hoşgörler Didem Keleş Serpil Tanrıverdi-akhisaroğlu Şeniz İnanç Mustafa Akhisaroğlu Ülker Cankurt Zekiye Aydoğdu Ahmet Deniz Uçar Oğuz Çetinayak Gülgün Oktay Sevil Gönenç Arda

Öz

Background: Matrix metalloproteinase (MMP) inhibitors decrease inflammation in normal tissues and suppress cancer progress in normal tissues. Valproic acid (VA) and doxycycline (DX) are MMP inhibitors that have radio-protective effects. Their ability to inhibit MMPs in irradiated tissue is unknown and the role of MMPs in radio-protective effects has not been tested to date. Aims: The purpose of this study was to examine whether administration of VA and DX to rats before irradiation affects tissue inflammation and apoptosis in the early phase of radiation, and whether the effect of these drugs is mediated by MMP inhibition. Study Design: Animal experimentation. Methods: Twenty-six Wistar rats were randomized into four groups: control (CTRL), radiation (RT), VA plus radiation (VA+RT), and DX plus radiation (DX+RT).Three study groups were exposed to a single dose of abdominal 10 Gy gamma radiation; the CTRL group received no radiation. Single doses of VA 300 mg/kg and DX 100 mg/kg were administered to each rat before radiation and all rats were sacrificed 8 hours after irradiation, at which point small intestine tissue samples were taken for analyses. Levels of inflammatory cytokines (TNF-α, IL-1β, and IL-6) and matrix metalloproteinases (MMP-2 and MMP 9) were measured by ELISA, MMP activities were measured by gelatin and casein zymography and apoptosis was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Results: VA decreased the levels of TNF-α and IL-1β proteins insignificantly and decreased apoptosis significantly in the irradiated tissue, but did not inhibit MMPs. In contrast, VA protected the basal MMP activities, which decreased in response to irradiation. No effect of DX was observed on the levels of inflammatory cytokines or activities of MMPs in the early phases of radiation apoptosis. Conclusion: Our findings indicated that VA protects against inflammation and apoptosis, and DX exhibits anti-apoptotic effects in early radiation and these effects are independent from MMP inhibition.

Anahtar Kelimeler

Matrix metalloproteinase, radiation, valproic acid, doxycycline

Kaynakça

• 1. Linard C, Ropenga A, Vozenin-Brotons M.C, Chapel A, Mathe D. Abdominal irradiation increases inflammatory cytokine expression and activates NF-kappaB in rat ileal muscularis layer. Am J Physiol Gastrointest Liver Physiol 2003;285:G556-65. [CrossRef]
• 2. Paris F., Fuks Z., Kang A., Capodieci P., Juan G., Ehleiter D., et al. Endothelial apoptosis as the primary lesion initiating intestinal radiation damage in mice. Science 2001;293:293-7. [CrossRef]
• 3. Angenete E., Langenskiöld M., Falk P., Ivarsson M.L. Matrix metalloproteinases in rectal mucosa, tumour and plasma: response after preoperative irradiation. Int J Colorectal Dis 2007;22:667-74. [CrossRef]
• 4. Strup-Perrot C., Vozenin-Brotons M.C., Vandamme M., Linard C., Mathé, D. Expression of matrix metalloproteinases and tissue inhibitor metalloproteinases increases in X-irradiated rat ileum despite the disappearance of CD8a T cells. World J Gastroenterol 2005;11:6312-21. [CrossRef]
• 5. Ito A., Mukaiyama A., Itoh Y., Nagase H., Thogersen I.B., Enghild JJ., et al. Degradation of interleukin 1beta by matrix metalloproteinases. J Biol Chem 1996;271:14657-60. [CrossRef]
• 6. Mohan MJ, Seaton T, Mitchell J, Howe A, Blackburn K, Burkhart W, et al. The tumor necrosis factor-alpha converting enzyme (TACE): a unique metalloproteinase with highly defined substrate selectivity. Biochemistry 2002;4:9462-9. [CrossRef]
• 7. Nascimento GC, Rizzi E, Gerlach RF, Leite-Panissi CR. Expression of MMP-2 and MMP-9 in the rat trigeminal ganglion during the development of temporomandibular joint inflammation. Braz J Med Biol Res 2013;46:956-67. [CrossRef]
• 8. Shen LC, Chen YK, Lin LM, Shaw SY. Anti-invasion and antitumor growth effect of doxycycline treatment for human oral squamous-cell carcinoma-in vitro and in vivo studies. Oral Oncol 2010;46:178-84. [CrossRef]
• 9. Lee JY, Kim HS, Choi HY, Oh TH, Ju BG, Yune TY. Valproic acid attenuates blood-spinal cord barrier disruption by inhibiting matrix metalloprotease-9 activity and improves functional recovery after spinal cord injury. J Neurochem 2012;121:818- 29. [CrossRef]
• 10. Papi A, Ferreri AM, Rocchi P, Guerra F, Orlandi M. Epigenetic modifiers as anticancer drugs: effectiveness of valproic acid in neural crest-derived tumor cells. Anticancer Res 2010;30:535-40.
• 11. Chung YL, Wang AJ, Yao LF. Antitumor histone deacetylase inhibitors suppress cutaneous radiation syndrome: Implications for increasing therapeutic gain in cancer radiotherapy. Mol Cancer Ther 2004;3:317-25.
• 12. Epperly MW, Chaillet JR, Kalash R, Shaffer B, Goff J, Franicola D, et al. Conditional radioresistance of Tet-inducible manganese superoxide dismutase bone marrow stromal cell lines. Radiat Res 2013;180:189-204. [CrossRef]
• 13. Panés J, Mollà M, Casadevall M, Salas A, Sans M, Conill C, et al. Tepoxalin inhibits inflammation and microvascular dysfunction induced by abdominal irradiation in rats. Aliment Pharmacol Ther 2000;14:841-50. [CrossRef]
• 14. Wang Z, Leng Y, Tsai LK, Leeds P, Chuang DM. Valproic acid attenuates blood-brain barrier disruption in a rat model of transient focal cerebral ischemia: the roles of HDAC and MMP-9 inhibition. J Cereb Blood Flow Metab 2011;31:52-7. [CrossRef]
• 15. Grant K. DOXYCYLINE. Rat Medication Guide. 2003 June 23 (cited 2010 January 19): Available from: http://ratguide.com/ meds/antimicrobial_agents/doxycycline.php
• 16. Heussen C, Dowdle EB. Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Anal Biochem 1980;102:196–202. [CrossRef]
• 17. Kerr JF, Winterford CM, Harmon BV. 1994. Apoptosis. Its significance in cancer and cancer therapy. Cancer 1994;73:2013- 26. [CrossRef]
• 18. Negoescu A, Lorimier P, Labat-Moleur F, Drouet C, Robert C, Guillermet C, et al. In situ apoptotic cell labeling by the TUNEL method: improvement and evaluation on cell preparations. J Histochem Cytochem 1996;44:959-68. [CrossRef]
• 19. Lee WH, Warrington JP, Sonntag WE, Lee YW. Irradiation alters MMP-2/TIMP-2 system and collagen type IV degradation in brain. Int J Radiat Oncol Biol Phys 2012;82:1559-66. [CrossRef]
• 20. Medina C, Radomski MW. Role of matrix metalloproteinases in intestinal inflammation. Pharmacol Exp Ther 2006;318:933-38. [CrossRef]
• 21. Hovdenak N, Wang J, Sung CC, Kelly T, Fajardo LF, Hauer-Jensen M. Clinical significance of increased gelatinolytic activity in the rectal mucosa during external beam radiation therapy of prostate cancer. Int J Radiat Oncol Biol Phys 2002;53:919-27. [CrossRef]
• 22. Kumar A, Collins HM, Scholefield JH, Watson SA. Increased type-IV collagenase (MMP-2 and MMP-9) activity following preoperative radiotherapy in rectal cancer. Br J Cancer 2000;82:960-5. [CrossRef]
• 23. Kim NY, Lee JE, Chang HJ, Lim CS, Nam DH, Min BH, et al. Gamma-irradiation enhances RECK protein levels in Panc-1 pancreatic cancer cells. Mol Cells 2008;25:105-11.
• 24. Lint VP, Libert C. Chemokine and cytokine processing by matrix metalloproteinases and its effect on leukocyte migration and inflammation. J Leukoc Biol 2007;82:1375-80. [CrossRef]
• 25. Manicone AM, McGuire JK. Matrix metalloproteinases as modulators of inflammation. Semin Cell Dev Biol 2008;19:34-41. [CrossRef]
• 26. Zhou Y, Niu J, Li S, Hou H, Xu Y, Zhang W, et al. Radioprotective effects of valproic acid, a histone deacetylase inhibitor, in the rat brain. Biomed Rep 2015;3:63-9.
• 27. Zhou Y, Xu Y, Wang H, Niu J, Hou H, Jiang Y. Histone deacetylase inhibitor, valproic acid, radiosensitizes the C6 glioma cell line in vitro. Oncol Lett 2014;7:203-8.
• 28. Jantzie LL, Todd KG. Doxycycline inhibits proinflammatory cytokines but not acute cerebral cytogenesis after hypoxia-ischemia in neonatal rats. J Psychiatry Neurosci 2010;35:20-32. [CrossRef]
• 29. Samtani S, Amaral J, Campos MM, Fariss RN, Becerra SP. Doxycycline-mediated inhibition of choroidal neovascularization. Invest Ophthalmol Vis Sci 2010;50:5098-106. [CrossRef