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Selective inhibition of cyclooxygenase‐2 protects porcine aortic endothelial cells from human antibody‐mediated complement‐dependent cytotoxicity
Author(s) -
Chen Pengfei,
Zhao Yanli,
Gao Hanchao,
Huang Jiabao,
Lu Ying,
Song Jinqi,
Lin Lizhong,
Lin Zejin,
Ou Chunpei,
Sun Huimin,
Li Yajing,
Zeng Changchun,
Cooper David K. C.,
Zhan Yongqiang,
Deng Xuefeng,
Mou Lisha
Publication year - 2019
Publication title -
xenotransplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.052
H-Index - 61
eISSN - 1399-3089
pISSN - 0908-665X
DOI - 10.1111/xen.12536
Subject(s) - propidium iodide , xenotransplantation , viability assay , proinflammatory cytokine , downregulation and upregulation , microbiology and biotechnology , tumor necrosis factor alpha , chemistry , biology , pharmacology , transplantation , programmed cell death , inflammation , immunology , apoptosis , medicine , biochemistry , gene
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.