目的 研究菥蓂Thlaspi arvense水提醇沉部位、正丁醇部位的抗炎活性以及正丁醇部位的作用機(jī)制。方法 將菥蓂水提物經(jīng)過乙醇、正丁醇萃取后制得菥蓂水提醇沉部位、正丁醇部位。采用MTT法確定水提醇沉部位、正丁醇部位（15.625、31.25、62.5、125、250、500、1 000 μg/mL）對RAW264.7細(xì)胞的毒性。采用脂多糖（LPS）1 μg/mL誘導(dǎo)RAW264.7細(xì)胞產(chǎn)生炎性反應(yīng)，建立體外炎癥細(xì)胞模型。Griess法檢測水提醇沉部位、正丁醇部位（7.812 5、15.625、31.25、62.5、125、250、500、1 000 μg/mL）對LPS誘導(dǎo)RAW264.7細(xì)胞后NO釋放量的影響；ELISA法檢測水提醇沉部位、正丁醇部位（125、250、500 μg/mL）對白細(xì)胞介素-6（IL-6）、腫瘤壞死因子-α（TNF-α）釋放量的影響；Western Blotting法檢測正丁醇部位（125、250、500 μg/mL）對環(huán)氧合酶-2（COX-2）、一氧化氮合酶（iNOS）、白細(xì)胞介素1β（IL-1β）、核因子-κB（NF-κB）p-P65、Toll樣受體4（TLR-4）、磷酸化NF-κB抑制蛋白（p-IκBα）蛋白表達(dá)的影響。結(jié)果 菥蓂水提醇沉部位、正丁醇部位質(zhì)量濃度在1 000 μg/mL以下時(shí)對RAW264.7細(xì)胞幾乎無毒性作用。與模型組比較，菥蓂水提醇沉部位、正丁醇部位62.5、125、250、500、1 000 μg/mL組NO釋放量顯著下降（P<0.01）；菥蓂水提醇沉部位、正丁醇部位125、250、500 μg/mL組IL-6、TNF-α的分泌量顯著降低（P<0.01），且正丁醇部位作用優(yōu)于水提醇沉部位；菥蓂正丁醇部位500 μg/mL組的iNOS和250、500 μg/mL組COX-2、IL-1β、NF-κB p-P65、TLR-4、p-IκBα蛋白表達(dá)量顯著降低（P<0.01），作用均呈濃度相關(guān)性。結(jié)論 菥蓂正丁醇部位顯著抑制LPS誘導(dǎo)的RAW264.7細(xì)胞NO的釋放，IL-1β、IL-6、TNF-α的分泌以及COX-2、iNOS蛋白的表達(dá)，其抗炎作用機(jī)制可能與抑制TLR-4/NF-κB信號(hào)通路相關(guān)。

Objective To study the anti-inflammatory activities of Thlaspi arvense water extraction and alcohol precipitation site and N-butanol site, and the mechanism of N-butanol site. Methods Thlaspi arvense water extracts were extracted by ethanol and N-butanol to obtain water extraction and alcohol precipitation site and N-butanol site. MTT assay was used to determine the toxicity of water extraction and alcohol precipitation site and N-butanol site (15.625, 31.25, 62.5, 125, 250, 500, 1 000 μg/mL) to RAW264.7 cells. RAW264.7 cells were induced by lipopolysaccharide (LPS) at 1 μg/mL, and the inflammatory cell model was established in vitro. Griess method was used to detect the effects of water extraction and alcohol precipitation site and N-butanol site (7.8125, 15.625, 31.25, 62.5, 125, 250, 500, 1 000 μg/mL) on NO release of RAW264.7 cells after LPS induction. ELISA method was used to detect the effects of water extraction and alcohol precipitation site and N-butanol site (125, 250, and 500 μg/mL) on the release of interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α). Western Blotting was used to detect the effects of N-butanol site (125, 250, and 500 μ g/mL) on the expression of cyclooxygenase-2 (COX-2), nitric oxide synthase (iNOS), interleukin 1β (IL-1β), NF- κ B P-P65, TLR-4 and P-IκBα. Results Thlaspi arvense water extraction and alcohol precipitation site and N-butanol site had almost no toxicity to RAW264.7 cells when the concentration was less than 1 000 μg/mL. Compared with model group, the release of NO in Thlaspi arvense water extraction and alcohol precipitation site and N-butanol site of 62.5, 125, 250, 500 and 1 000 μg/mL groups was significantly decreased (P<0.01). The secretion levels of IL-6 and TNF-α were significantly decreased (P<0.01) in Thlaspi arvense water extraction and alcohol precipitation site and N-butanol site of 125, 250 and 500 μg/mL, and the effects of N-butanol were better than those of water extraction and alcohol precipitation site. The protein expression levels of iNOS in 500 μg/mL and COX-2, IL-1β, NF-κB p-P65, TLR-4, and p-IκBα in 250 and 500 μg/mL groups of N-butanol site decreased significantly (P<0.01), and the effects were correlated with concentration. Conclusion Thlaspi arvense N-butanol site was almost non-cytotoxic, inhibited the release of RAW264.7 cell NO, inhibited the secretion of inflammatory factor IL-1β, IL-6, TNF-α and the expression of COX-2、iNOS protein in a dose-dependent manner. The anti-inflammatory effect of extract may be mediated by inhibition of TLR-4/NF-κB signaling pathway.

R285.5

國家自然科學(xué)基金項(xiàng)目（21262005）；廣西科技重大專項(xiàng)項(xiàng)目（桂科AA18242040）