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Rhein antagonizes P2X7 receptor in rat peritoneal macrophages.

ABSTRACT: P2X7 receptor plays important roles in inflammation and immunity, and thereby it serves as a potential therapeutic target for inflammatory diseases. Rhein, an anthraquinone derivative, exhibits significant anti-inflammatory and immunosuppressive activities in therapy. However, the underlying mechanisms are largely unclear. Here, we aimed to investigate the effects of rhein on P2X7 receptor-mediated responses in vitro. In HEK293 cells expressing rat P2X7 receptor, we first found that rhein concentration-dependently blocked ATP-induced cytosolic calcium concentration ([Ca(2+)]c) elevation and pore formation of the plasma membrane, two hallmarks of the P2X7 receptor activation. These two inhibitory effects of rhein were also observed in rat peritoneal macrophages. Furthermore, rhein counteracted macrophage phagocytosis attenuation and suppressed reactive oxygen species (ROS) production triggered by ATP/BzATP. Meanwhile, rhein reduced ATP/BzATP-induced IL-1? release in lipopolysaccharide-activated macrophages. Prolonged application of ATP caused macrophage apoptosis, while the presence of rhein suppressed this cell cytotoxicity. Such ATP/BzATP-induced cellular reactions were also inhibited by a well-known rat P2X7 receptor antagonist, brilliant blue G, in a similar way to rhein. Together, our results demonstrate that rhein inhibit ATP/BzATP-induced [Ca(2+)]c increase, pore formation, ROS production, phagocytosis attenuation, IL-1? release and cell apoptosis by antagonizing the P2X7 receptor in rat peritoneal macrophages.

SUBMITTER: Hu F

PROVIDER: S-EPMC4564849 | biostudies-other | 2015

REPOSITORIES: biostudies-other

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Rhein antagonizes P2X7 receptor in rat peritoneal macrophages.

Hu Fen F Xing Fulin F Zhu Ge G Xu Guangxue G Li Cunbo C Qu Junle J Lee Imshik I Pan Leiting L

Scientific reports 20150910

P2X7 receptor plays important roles in inflammation and immunity, and thereby it serves as a potential therapeutic target for inflammatory diseases. Rhein, an anthraquinone derivative, exhibits significant anti-inflammatory and immunosuppressive activities in therapy. However, the underlying mechanisms are largely unclear. Here, we aimed to investigate the effects of rhein on P2X7 receptor-mediated responses in vitro. In HEK293 cells expressing rat P2X7 receptor, we first found that rhein concen ...[more]

PMID: 26354875

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Project description:Diabetic cardiomyopathy (DCM) is a critical complication of long-term chronic diabetes mellitus, and it is characterized by myocardial fibrosis and myocardial hypertrophy. Previous studies have shown that the pyroptosis pathway was significantly activated in DCM and may be related to the P2X7 receptor. However, the role of the P2X7 receptor in the development of DCM with pyroptosis is still unclear. In this study, we aimed to explore the mechanism of puerarin and whether the P2X7 receptor can be used as a new target for puerarin in the treatment of DCM. We adopted systematic pharmacology and bioinformatic approaches to identify the potential targets of puerarin for treating DCM. Additionally, we employed D-glucose-induced H9C2 rat cardiomyocytes and lipopolysaccharide-treated RAW264.7 mouse mononuclear macrophages as the in vitro model on DCM research, which is close to the pathological conditions. The mRNA expression of cytokines in H9C2 cells and RAW264.7 macrophages was detected. The protein expressions of NLRP3, N-GSDMD, cleaved-caspase-1, and the P2X7 receptor were investigated with Western blot analysis. Furthermore, molecular docking of puerarin and the P2X7 receptor was conducted based on CDOCKER. A total of 348 puerarin targets and 4556 diabetic cardiomyopathy targets were detected, of which 218 were cross targets. We demonstrated that puerarin is effective in enhancing cardiomyocyte viability and improving mitochondrial function. In addition, puerarin is efficacious in blocking NLRP3-Caspase-1-GSDMD-mediated pyroptosis in H9C2 cells and RAW264.7 cells, alleviating cellular inflammation. On the other hand, similar experimental results were obtained by intervention with the P2X7 receptor antagonist A740003, suggesting that the protective effects of puerarin are related to the P2X7 receptor. The molecular docking results indicated key binding activity between the P2X7 receptor and puerarin. These findings indicate that puerarin effectively regulated the pyroptosis signaling pathway during DCM, and this regulation was associated with the P2X7 receptor.

Project description:ObjectiveTo investigate the effects of AMPK activation on mitochondrial inhibition by uremic serum through the AMPK-activated rat peritoneal macrophages stimulated by uremic serum, thereby providing a reference for the clinical treatment of chronic kidney disease.MethodsTwenty-two male Sprague-Dawley (SD) rats were included as experimental subjects. Fifteen rats were constructed into chronic kidney disease models (the model group). The remaining seven rats only received renal capsule stripping instead of nephrectomy (the sham-operated group). Ten weeks after model construction, the bodyweight, blood biochemical indicators, and metabolic parameters of rats in groups were measured. Meanwhile, the expression of the M1 phenotype marker protein in peritoneal macrophages was determined.ResultsTen weeks after model construction, the bodyweight of rats in the model group was significantly lower than that in the sham-operated group. The values of urea nitrogen and serum creatinine were significantly higher than those in the sham-operated group (P<0.01). The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and the monocyte chemoattractant protein 1 (MCP-1) of rats in the model group were significantly higher than those in the sham-operated group (P <0.01). After the lipopolysaccharide (LPS) stimulation, the expressions of M1 phenotype marker mRNA in the model group was significantly increased. The expression of mitochondrial structural protein mRNA in the peritoneal macrophages of rats in the model group was significantly lower than that in the sham-operated group. The expression of M1 phenotype marker mRNA was significantly decreased in the uremic serum group after AMPK agonist (P<0.01).ConclusionIn rats with chronic renal insufficiency, mitochondrial regeneration was dysfunctional in macrophages. By activating AMPK, the inhibitory effect of uremia serum on mitochondrial regeneration of macrophages was improved. Therefore, AMPK was a critical factor that could regulate mitochondrial regeneration of macrophages.

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Rhein antagonizes P2X7 receptor in rat peritoneal macrophages.

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Rhein antagonizes P2X7 receptor in rat peritoneal macrophages.

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