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【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 (SIN) on endotoxemia induced by bacterial endotoxin (LPS) and the expression of A2A and P2X7 in puromycin-induced macrophage RAW264.7 cells. 【Methods】 BALB / c mice were randomly divided into blank control group, model group and sinomenine group (doses of 40, 80 and 160 mg / kg, respectively). Sinomenine group was administered intraperitoneal injection of sinomenine for 30 min. Group and sinomenine group were given intraperitoneal injection of LPS (15 mg / kg dose) to establish a mouse model of endotoxemia, the level of serum tumor necrosis factor-α (TNF-α) was measured by enzyme-linked immunosorbent assay (ELISA) , Interleukin-6 (IL-6), and the expression of purine receptors A2A and P2X7 in liver and spleen by reverse transcription-polymerase chain reaction (RT-PCR) LPS group, sinomenine group and sinomenine group were treated with sinomenine (300μmol / L) for 2 h, LPS group and sinomenine group were given LPS (100 ng / m L). After cultured for 8 h, the secretion of TNF-α in supernatant was detected by ELISA. The expression of P2X7 and A2A m RNA in cells was detected by RT-PCR. 【Results】 The levels of TNF-α and IL-6 in serum of mice were significantly increased after LPS stimulation. The expressions of A2A and P2X7 in liver and spleen were significantly increased. Sinomenine could down-regulate the expressions of TNF-α and IL- 6, and inhibited the expression of A2A and P2X7 (all P <0.05). Under the stimulation of LPS in vitro, the expression of TNF-α and A2A and P2X7 were increased in RAW264.7 cells. Sinomenine could inhibit the secretion of TNF- And down-regulated the expression of P2X7 (all P <0.05), but promoted the expression of A2A. 【Conclusion】 Sinomenine can inhibit the expression of purine receptor P2X7 in the liver, spleen and macrophage of LPS-stimulated mice, while the effect of sinomenine on the expression of A2A in different tissues is different.