摘要
【目的】探讨青藤碱(sinomenine,SIN)对细菌内毒素(LPS)刺激的小鼠内毒素血症模型及巨噬细胞RAW264.7细胞炎症模型嘌呤受体A2A、P2X7表达的影响。【方法】选用BALB/c小鼠设空白对照组,模型组,青藤碱组(剂量分别为40、80、160 mg/kg),青藤碱组给予腹腔注射青藤碱30 min后,模型组和青藤碱组给予腹腔注射LPS(剂量为15 mg/kg)建立内毒素血症小鼠模型,采用酶联免疫吸附(ELISA)法检测小鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平;逆转录—聚合酶链反应(RT-PCR)检测肝脏、脾脏嘌呤受体A2A、P2X7的表达。RAW264.7细胞设空白组、LPS组、青藤碱组,青藤碱组给予青藤碱(300μmol/L)干预2 h后,LPS组和青藤碱组均给予LPS(剂量为100 ng/m L),继续培养8 h后,采用ELISA法检测上清TNF-α分泌量,RT-PCR法检测细胞P2X7、A2A m RNA表达。【结果】在LPS刺激下,小鼠血清TNF-α和IL-6水平显著上升,肝脏、脾脏组织中嘌呤受体A2A、P2X7的表达显著升高,青藤碱能下调TNF-α和IL-6水平,并抑制A2A、P2X7的表达(均P<0.05);体外LPS刺激下,RAW264.7细胞分泌TNF-α增加,A2A、P2X7的表达增加,青藤碱可抑制TNF-α的分泌,并下调P2X7的表达(均P<0.05),但对A2A的表达有促进作用。【结论】青藤碱对LPS刺激的小鼠肝脏、脾脏及体外巨噬细胞中嘌呤受体P2X7的表达增加均表现抑制作用,而对不同组织细胞中A2A表达的影响不同,相关机制有待深入。
Objective To investigate the effect of sinomenine on the purinergic receptors A2 A 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 A2 A and P2X7 in the liver and spleen were detected by reverse transcription- polymerase chain reaction（RT-PCR）. RAW264.7macrophages were divided into blank control group,LPS group and sinomenine group. Sinomenine group was firstly treated with sinomenine（300 μmol/L）for 2 h,and then LPS group and sinomenine group were treated with LPS（100 ng/m L）for another 8 hours. TNF-α in the cell supernatant was measured by ELISA,and the expression levels of A2 A 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 A2 A 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 A2 A and P2X7 in RAW264.7cells, and sinomenine decreased TNF- α content and P2X7expression（P〈 0.05）, but had an effect on enhancing A2 A 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 A2 A in various tissues and cells varies,whose related mechanism is needed further study.
出处
《广州中医药大学学报》
CAS
2016年第1期97-103,共7页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
国家自然科学基金项目(编号:81573672)
关键词
青藤碱/药理学
嘌呤受体
脂多糖
细胞因子
基因表达调控
细胞培养
疾病模型
动物
小鼠
sinomenine/pharmacology
purinergic receptors
lipopolysaccharide
cytokines
gene expression regulation
cell culture
disease models
animal
mice
