Effect of Sinomenine on Expression of Purinergic Receptors A2A and P2X7 in Mouse Model and In-vitro Macrophages Stimulated by Lipopolysaccharide
10.13359/j.cnki.gzxbtcm.2016.01.024
- VernacularTitle:青藤碱对细菌内毒素刺激的小鼠及巨噬细胞嘌呤受体A2A、 P2X7表达的影响
- Author:
Jing LI
;
Yangyang WU
;
Haisong ZHOU
;
Ruili ZHU
;
Lang YI
;
Yan DONG
;
Peixun WANG
- Publication Type:Journal Article
- Keywords:
sinomenine/pharmacology;
purinergic receptors;
lipopolysaccharide;
cytokines;
gene expression regulation;
cell culture;
disease models,animal;
mice
- From:
Journal of Guangzhou University of Traditional Chinese Medicine
2016;(1):97-103
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of sinomenine on the purinergic receptors A2A and P2X7 in endotoxemia mouse model and RAW264.7 macrophage model stimulated by lipopolysaccharide(LPS). Methods BALB/c mice were randomly divided into blank control group, model group and sinomenine group. Thirty minutes after the rats of sinomenine group were pretreated with intraperitoneal injection of sinomenine (40, 80, 160 mg/kg), the mice were given intraperitoneal injection of 15 mg/kg LPS to induce endotoxemia model. The serum levels of tumor necrosis factor-alpha(TNF-α) and interleukin-6(IL-6) were measured by enzyme-linked immunosorbent assay (ELISA). The expression levels of purinergic receptor A2A and P2X7 in the liver and spleen were detected by reverse transcription-polymerase chain reaction(RT-PCR). RAW264.7 macrophages were divided into blank control group, LPS group and sinomenine group. Sinomenine group was firstly treated with sinomenine(300μmol/L) for 2 h, and then LPS group and sinomenine group were treated with LPS (100 ng/mL) for another 8 hours. TNF-α in the cell supernatant was measured by ELISA, and the expression levels of A2A and P2X7 in RAW264.7 cells were detected by RT-PCR. Results Stimulation with LPS could induce the increase of the mouse serum levels of TNF-α and IL-6 as well as the expression of A2A and P2X7 in mouse liver and spleen, and sinomenine had a counteraction on the above indexes(P<0.05) . In-vitro stimulation with LPS could induce the increase of the content of TNF-α and the expression of A2A and P2X7 in RAW264.7 cells , and sinomenine decreased TNF-α content and P2X7 expression (P<0.05) , but had an effect on enhancing A2A expression. Conclusion Sinomenine suppresses the expression of purinergic receptor P2X7 in mouse spleen and liver as well as in RAW264.7 macrophages, but its effect on the expression of A2A in various tissues and cells varies, whose related mechanism is needed further study.
