青藤碱通过糖皮质激素受体发挥抗炎作用的研究  

Anti-inflammation Effects of Sinomenine Through Blocking and Silencing Glucocorticoid Receptor

作　　者：易浪[1] 朱瑞丽[1] 谢冰冰[1] 王青[1] 董燕[1] 

机构地区：[1]广州中医药大学临床药理研究所免疫研究室,广东广州510405

出　　处：《中药新药与临床药理》2015年第4期494-499,共6页Traditional Chinese Drug Research and Clinical Pharmacology

摘　　要：目的观察阻断及沉默Raw264.7细胞糖皮质激素受体(GR)对青藤碱(SIN)抗炎作用的影响。方法采用小鼠巨噬细胞系Raw264.7为研究对象,以脂多糖(LPS)诱导细胞炎症模型,选择糖皮质激素药地塞米松(Dex)作为阳性药,选用GR特异性拮抗剂米非司酮(Ru486)进行干预,并采用si RNA技术沉默GR。ELISA检测细胞分泌肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的变化,Western blot检测GR的表达,实时荧光定量PCR(q RT-PCR)检测TNF-α、IL-6 m RNA水平的变化。结果 q RT-PCR和ELISA检测结果显示LPS刺激下Raw264.7细胞的TNF-α与IL-6在m RNA和蛋白水平均升高(P<0.05),Dex和SIN作用后均可明显降低其表达(P<0.05),但在GR拮抗剂干预下,Dex和SIN的抗炎作用被抑制,与LPS组比较,差异无统计学意义(P>0.05)。炎症状态下,GR蛋白表达较正常组明显降低(P<0.05),Dex和SIN对GR蛋白表达的影响不明显(P>0.05)。si RNA技术成功沉默GR蛋白表达后,Dex和SIN的抗炎作用明显被减弱(P<0.05)。结论SIN能通过作用于GR发挥抗炎作用,具有类皮质激素样作用,GR是SIN发挥抗炎作用的靶点之一。Objective To observe the influence of blocking and silencing glucocorticoid receptor （GR） on the anti-inflammatory effects of sinomenine （SIN）. Methods Mouse macrophage cell strain Raw264.7 was stimulated by lipopolysaccharide to induce cell inflammation model, dexamethasone （Dex） was used as the positive drug, and mifepristone （Ru486） was used as the blocker, siRNA interference technology was used for silencing GR, and then ELISA was applied for detecting cell secretion of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6）. Western blotting was used for detecting GR expression, and real-time fluorescent quantitation PCR was applied for the detection of TNF- α and IL-6 mRNA levels. Results The results of ELISA and real-time fluorescent quantitation PCR showed that the mRNA and protein production of TNF-α and IL-6 induced by LPS was increased （P 〈 0.05 ）, and Dex and SIN reduced the production significantly （P 〈 0.05）. But after the intervention with antagonist Ru486, the anti- inflammatory effect of Dex and SIN were blocked, and the difference was insignificant compared with the LPS group（P 〉 0.05 ）. At inflammatory state, the expression of GR was decreased significantly（P 〈 0.05 ） compared with the normal group, and treatment of Dex and SIN showed less effect on the expression of GR（P〉 0.05）. After GR siRNA intervention, GR expression was silenced obviously, and the anti-inflammatory effects of Dex and SIN were also be reduced （P 〈 0.05 ）. Conclusion Sinomenine exerts anti-inflammatory effects by acting on GR, which shows corticosteroid-like effect. Glucocorticoid receptor is one of the targets of sinomenine to play an anti-inflammatory role.

关 键 词：青藤碱 糖皮质激素受体 炎症 肿瘤坏死因子-Α 白细胞介素-6 

分 类 号：R285.5[医药卫生—中药学]