Ketanserin, a serotonin 2A receptor antagonist, alleviates ischemia-related biliary fibrosis following donation after cardiac death liver transplantation in rats

Abstract

Biliary fibrosis is a major complication after donation after cardiac death (DCD) liver transplantation. In this process, the roles of serotonin [5-hydroxytryptamine (5-HT)] and the 5-HT2A receptor subtype are still unknown. In this study, we analyzed markers of portal fibroblast (PF)/myofibroblast (MF) transdifferentiation such as transforming growth factor b1 (TGF-b1), phosphorylated smad2/3, a-smooth muscle actin (a-SMA), collagen I, and collagen III in a primary culture system of PFs after the administration of 5-HT or 5-HT plus ketanserin (a selective 5-HT2A receptor antagonist). A rat DCD transplant model was established with 30 minutes of warm ischemia and 4 hours of cold ischemia during organ procurement. Recipients were intraperitoneally injected with ketanserin (1 mgÁkg 21 Áday 21 ) or normal saline. Grafts without in situ warm ischemia instead of minimal cold storage (30 minutes) served as controls. The serum biochemistry, the liver contents of 5-HT and hydroxyproline (HYP), and the expression of fibrosis-related genes (including TGF-b1, matrix metalloproteinase 2, procollagen a1, and a-SMA messenger RNA) were determined. The extent of biliary fibrosis was also assessed histopathologically. The results indicated that ketanserin inhibited 5-HT-activated TGF-b1-smad2/3 signaling in vitro and thereby depressed the MF conversion of PFs. Rats receiving DCD livers showed increased liver contents of 5-HT and HYP, impaired biliary function, up-regulation of fibrosis-related genes, and aggravated biliary fibrosis. However, these phenomena were alleviated by treatment with ketanserin. We concluded that the profibrotic activity of 5-HT occurred through the activation of TGF-b1 signaling and the 5-HT2A receptor. Thus, these data suggest that the 5-HT2A receptor may be a potential therapeutic target for ischemia-related biliary fibrosis after DCD liver transplantation. Liver Transpl 20:1317-1326, 2014. V C 2014 AASLD.Received March 30, 2014; accepted July 2, 2014.The worldwide organ shortage has led to the increased use of donation after cardiac death (DCD) livers. Because of the additional warm ischemia before preservation, DCD liver grafts are at higher risk for failure and bile duct injury. Currently, DCD liver grafts are the most common cause of nonanastomotic biliary strictures, which are characterized as biliary strictures and dilatation at any location in the biliary system of the transplanted liver.1,2 They are associated with an increased incidence of biliary sludge, recurrent Abbreviations: 5-HT, 5-hydroxytryptamine; 5-HT1K, 5-hydroxytryptamine and ketanserin; a-SMA, a-smooth muscle actin; ALP, alkaline phosphatase; BECs, biliary epithelial cells; DCD, donation after cardiac death; GGT, gamma-glutamyl transpeptidase; HSC, hepatic stellate cell; HYP, hydroxyproline; MF, myofibroblast; MMP2, matrix metalloproteinase 2; mRNA, messenger RNA; PF, portal fibroblast; PI, proliferation index; p-smad, phosphorylated drosophila mothers against decapentaplegic protein; RTqPCR, quantitative real-time reverse-transcr...

Stimulation of Anxiety-Like Behavior via ERK Pathway by Competitive Serotonin Receptors 2A and 1A in Post-Traumatic Stress Disordered Mice

“…Several studies have suggested anxiety-like behavior to be most often than not accompanied by protein expressional changes in 5HT2A and 5HT1A receptors in mice hippocampi [40, 46, 47]. Our western blot and immunofluorescence labeling results showed significant increment in the expression level of 5HT1A receptor after PTSD model establishment.…”

The Role of Serotonin during Skin Healing in Post-Thermal Injury

“…Having low fibroblasts and increased proportion of myofibroblasts supports our findings of impaired re-epithelialization with ketanserin and fluoxetine administration. Our results conflict with a previous study that suggests systemic ketanserin administration causes down regulation of α-SMA expression in portal fibroblasts pre-treated with serotonin [ 40 ]. However, this study evaluated the mRNA and protein expression of cultured portal fibroblasts, while our observation was based on quantifying α-SMA-positive cells using skin tissue immunostaining.…”

“…Therefore, it is possible that the increased wound area and impaired epithelialization following ketanserin treatment is due to the impairment of the inflammatory response. Our results showing suppression of fibroblast and keratinocyte proliferation with ketanserin treatment also support previous studies indicating that ketanserin reduces fibroblast proliferation and induce fibrosis in the liver [ 40 ], whereas it is also documented that activation of 5HTR2A signaling promotes fibrosis in various other tissues [ 16 , 33 , 34 ].…”

Application value of dual-energy computed tomography spectrum curve combined with clinical risk factors in predicting adherent perinephric fat

“…Activation of a chronic inflammation system can cause endothelial cells to secrete a variety of cytokines, chemokines, and plasminogen activator inhibitor type 1 (PAI-1), thereby reducing the fibrinolytic activity and leading to fibrous adhesion between perirenal fat and the renal capsule (17,18). In addition, serotonin [5-hydroxytryptamine (5-HT)], considered as fibrosis-related growth factor which is highly expressed in renal cell carcinoma contributes to APF (19,20). These paracrine mechanisms may partially explain why the probability of APF in benign renal tumors was lower than that in malignant tumors in our study.…”