南蛇藤素可抑制肿瘤坏死因子α诱导RAW264.7细胞的炎症及增殖

Celastrol inhibits tumor necrosis factor-alpha induced proliferation and inflammatory responses in RAW264.7 cells

背景:中药南蛇藤具有祛风活血、消肿止痛作用,而其有效单体成分南蛇藤素可能是其主要作用成分之一。目的:进一步验证南蛇藤素对肿瘤坏死因子α诱导RAW264.7细胞炎症和增殖的作用。方法:用0,1,10和100μg/L肿瘤坏死因子α刺激RAW264.7细胞建立体外炎症模型,用0.1,0.5,1.0,2.0μmol/L的南蛇藤素作用于该模型,ELISA法检测RAW264.7细胞白细胞介素1β,6,8和前列腺素E2分泌能力,CCK-8比色法检测RAW264.7细胞存活率,硝酸还原酶法检测炎性递质一氧化氮生成,Real-time PCR检测诱导型一氧化氮合酶,cyclin D1和cyclin E1基因m RNA表达水平。结果与结论:(1)以0.1、10和100μg/L肿瘤坏死因子α孵育RAW264.7细胞后,与对照组(0μg/L)比较,能不同程度促进RAW264.7细胞白细胞介素1β,6,8和前列腺素E2的分泌,增强一氧化氮的释放,促进细胞增殖并增加诱导型一氧化氮合酶,细胞周期蛋白D1和细胞周期蛋白E1的mR NA表达水平(P<0.05),其中10μg/L肿瘤坏死因子α对细胞的作用最强;(2)0.4μmol/L南蛇藤素能明显抑制肿瘤坏死因子α诱导的RAW264.7细胞白细胞介素1β,6,8和前列腺素E2的分泌,降低一氧化氮的释放,抑制细胞增殖并下调诱导型一氧化氮合酶,细胞周期蛋白D1和细胞周期蛋白E1的mR NA表达水平(P<0.05);(3)结果提示,南蛇藤素能够抑制肿瘤坏死因子α诱导的RAW264.7细胞的炎症反应和增殖能力。

BACKGROUND： Celastrol is one of the active components extracted from the traditional Chinese medicine Celastrus orbiculatus characterized by expelling the wind and promoting blood circulation and relieving swelling and pain.OBJECTIVE： To investigate the effects of celastrol on tumor necrosis factor-α（TNF-α） induced proliferation and inflammatory responses in RAW264.7 cells. METHODS： In vitro inflammatory cell models induced by TNF-α（0, 1, 10, 100 μg/L） were treated with celastrol（0.1, 0.5, 1.0, 2.0 μmol/L）. Interleukin-1β,-6,-8 and prostaglandin E2 in the models were measured by enzyme-linked immunosorbent assay. Cell survival was determined by cell counting kit-8. The inflammatory mediator nitric oxide secretion was detected by nitrate reductase assay. mR NA expressions of inducible nitric oxide synthase, cyclin D1 and cyclin E1 were detected by real-time polymerase chain reaction technique. RESULTS AND CONCLUSION： Significantly increased secretion of interleukin-1β,-6,-8, prostaglandin E2 and nitric oxide, cell proliferation, and m RNA expressions of inducible nitric oxide synthase, cyclin D1 and cyclin E1 were observed after the induction of TNF-α（0.1, 10, 100 μg/L） compared with the control group（without the induction of TNF-α）（P〈0.05）; especially, 10 μg/L of TNF-α exhibited the strongest effects. 0.4 μmol/L celastrol significantly suppressed TNF-α-induced release of interleukin-1β,-6,-8, prostaglandin E2 and nitric oxide, cell proliferation, and m RNA expressions of inducible nitric oxide synthase, cyclin D1 and cyclin E1 in RAW264.7 cells（P〈0. 05）. Our results demonstrate that celastrol can inhibit TNF-α-induced inflammatory response and proliferation in RAW264.7 cells.