Selective inhibition of cyclooxygenase‐2 protects porcine aortic endothelial cells from human antibody‐mediated complement‐dependent cytotoxicity
| Autor: | Changchun Zeng, Ying Lu, Jiabao Huang, David K. C. Cooper, Lin Lizhong, Chunpei Ou, Yongqiang Zhan, Jin-qi Song, Huimin Sun, Chen Pengfei, Lisha Mou, Yanli Zhao, Gao Hanchao, Deng Xuefeng, Ze-jin Lin, Li Yajing |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Small interfering RNA Swine Xenotransplantation medicine.medical_treatment Transplantation Heterologous Immunology Apoptosis 030230 surgery 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Downregulation and upregulation In vivo medicine Animals Propidium iodide Aorta Inflammation Transplantation biology Tumor Necrosis Factor-alpha Chemistry NF-kappa B Endothelial Cells Molecular biology Complement-dependent cytotoxicity 030104 developmental biology Cyclooxygenase 2 biology.protein Cytokines Tumor necrosis factor alpha Cyclooxygenase |
| Zdroj: | Xenotransplantation. 26 |
| ISSN: | 1399-3089 0908-665X |
| DOI: | 10.1111/xen.12536 |
| Popis: | Background Cyclooxygenase-2 (COX-2) is an inducible enzyme with catalytic activity for biosynthesis of prostaglandins which are the key mediators of inflammation. COX-2 is also the therapeutic target for widely used non-steroidal anti-inflammatory drugs (NSAIDs). However, the involvement of COX-2 in xenotransplantation (eg, pig-to-non-human primate) remains poorly recognized. Methods We investigated the mechanisms that regulate COX-2 expression and the effects of COX-2 on porcine aortic endothelial cell (PAEC) viability using in vitro pig-to-primate xenotransplantation model and in vivo pig-to-mouse cellular transplant model. Regulation of COX-2 expression was assessed by real-time quantitative polymerase chain reaction (qPCR) and Western blotting. The effects of inhibition or downregulation of COX-2 on PAEC viability were assessed by propidium iodide (PI)-Annexin V staining and Cell Counting Kit-8 assay. Results Human serum triggered robust COX-2 expression in PAECs in a dose- and time-dependent manner. Induction of COX-2 expression by human serum was partially through activation of both canonical and non-canonical nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κb) signaling and increasing intracellular calcium. Cytokines like tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), IL-17, were able to induce COX-2 expression. Selective inhibition of COX-2 by celecoxib dramatically decreased PAEC death in vitro and in vivo as defined by propidium iodide (PI)-Annexin V staining. Consistently, downregulation of COX-2 expression by NF-κb inhibitors or calcium chelator BAPTA decreased human serum-induced PAEC death as well. Silencing of COX-2 expression by small interfering RNA (siRNA) protected PAEC viability when transplanted under kidney capsule of C57BL/6 mice. Conclusions Taken together, our data suggest that COX-2 is highly induced in PAECs by xenogenic serum and associated with human antibody-mediated complement-dependent cytotoxicity. COX-2 might be a potential therapeutic target to improve xenotransplantation. |
| Databáze: | OpenAIRE |
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