Therapeutic value of melatonin post-treatment on CCl4-induced fibrotic rat liver
Publication: Canadian Journal of Physiology and Pharmacology
20 July 2015
Abstract
Melatonin is known for being beneficial in targeting liver diseases. This study aimed to investigate whether melatonin post-treatment is capable of rat carbon tetrachloride (CCl4)-induced liver fibrosis reduction. Thirty-two male Sprague-Dawley rats were divided into 4 groups: normal; fibrosis with CCl4 injection (1 mL/kg) twice weekly for 8 weeks; phosphate-buffered saline (PBS); and melatonin (20 mg/kg) for a further 4 weeks on cessation of CCl4. At the beginning of week 13, liver tissue samples were used for hematoxylin-eosin (H&E), periodic acid-Schiff (PAS), Masson’s trichrome (MT), and Oil Red O staining, quantitative real-time PCR (qRT-PCR) analysis of the matrix metalloproteinase-9 (MMP-9), MMP-13, transforming growth factor-β1 (TGF-β1), Bcl-2, and Bax genes as well as immunofluorescence (IF) of the first 3, and sera for measurement of aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin, and hydroxyproline. Chronic administration of CCl4 followed by considerable increase in tissue disruption, macro- and micro-vesicles, collagen, lipid droplets (LDs), AST, ALT, hydroxyproline, TGF-β1, and Bax, and decrease in glycogen depository, albumin, Bcl-2, MMP-9, and MMP-13; however, the pattern was reverse when it comes to melatonin treatment (for all p < 0.05). Our results reveal the beneficial aspects of melatonin in treatment of liver fibrosis probably via inhibition of TGF-β1expression.
Résumé
La mélatonine est connue pour être bénéfique en ciblant les maladies hépatiques. Cette étude visait à déterminer si un post-traitement à la mélatonine est capable de réduire la fibrose hépatique induite par le tétrachlorure de carbone (CCl4) chez le rat. Trente-deux rats mâles Sprague-Dawley ont été répartis en quatre groupes : normal; fibrose avec une injection de CCl4 (1 ml/kg) deux fois par semaine pendant huit semaines; solution saline tamponnée au phosphate (PBS); mélatonine (20 mg/kg) pendant quatre semaines supplémentaires après la cessation du CCl4. Au début de la treizième semaine, des échantillons de tissu hépatique ont été colorés à l’hématoxyline-éosine (H&E), à l’acide periodique couplé au réactif de Schiff (PAS), au trichrome de Masson (MT) et au Oil Red O; ils ont été soumis à une PCR quantitative (qRT-PCR) de la métalloprotéase matricielle-9 (MMP 9), de la MMP 13, du facteur de croissance transformant-β1 (TGF-β1), de Bcl-2 et Bax, de même qu’à une immunofluorescence (IF) afin de mesurer les trois premiers marqueurs; l’aspartate aminotransférase (AST), l’alanine aminotransférase (ALT), l’albumine et l’hydroxyproline du sérum ont aussi été mesurées. L’administration chronique de CCl4 était suivie d’une augmentation considérable de dommages tissulaires, de macro- et de microvésicules, de collagène, de gouttelettes lipidiques, d’AST, d’ALT, d’hydroxyproline, de TGF-β1, et de Bax, ainsi que d’une diminution du dépôt de glycogène, d’albumine, de Bcl-2, de MMP 9 et de MMP 13; cependant, ce patron était renversé par le traitement à la mélatonine (p < 0.05, pour tous). Les résultats des auteurs révèlent les aspects bénéfiques de la mélatonine dans le traitement de la fibrose hépatique probablement par l’intermédiaire de l’inhibition de l’expression de TGF-β1. [Traduit par la Rédaction]
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Published In
Canadian Journal of Physiology and Pharmacology
Volume 94 • Number 2 • February 2016
Pages: 119 - 130
History
Received: 14 June 2015
Accepted: 29 June 2015
Accepted manuscript online: 20 July 2015
Version of record online: 20 July 2015
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© 2016.
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Keywan Mortezaee, Fatemeh Sabbaghziarani, Ameneh Omidi, Ahmad Reza Dehpour, Negar Omidi, Soudabeh Ghasemi, Parichehr Pasbakhsh, and Iraj Ragerdi Kashani. 2016. Therapeutic value of melatonin post-treatment on CCl4-induced fibrotic rat liver. Canadian Journal of Physiology and Pharmacology. 94(2): 119-130. https://doi.org/10.1139/cjpp-2015-0266