[關(guān)鍵詞]
[摘要]
目的 香附抗神經(jīng)炎癥活性部位確定及成分分析。方法 基于脂多糖(LPS)誘導(dǎo)BV2細(xì)胞炎癥模型,Griess法檢測培養(yǎng)上清液中NO水平評價香附95%乙醇提取物(CR-95E,6.25、12.50、25.00、50.00 μg · mL-1)、50%乙醇提取物(CR-50E,6.25、12.50、25.00、50.00、100.00 μg·mL-1)、水提取物(CR-W,6.25、12.50、25.00、50.00、100.00 μg·mL-1)的抗炎活性;ELISA試劑盒法考察活性部位對模型細(xì)胞上清液中腫瘤壞死因子-α(TNF-α)、白細(xì)胞介素-1β(IL-1β)和白細(xì)胞介素-6(IL-6)含量的影響;采用ELISA法檢測上清液中犬尿氨酸途徑激活關(guān)鍵酶吲哚胺2,3-雙加氧酶(IDO)的含量變化;采用氣相色譜-質(zhì)譜聯(lián)用(GC-MS)技術(shù)對CR-95E進行化學(xué)成分定性分析,與NIST MS search 2.0質(zhì)譜檢索數(shù)據(jù)庫比對鑒定化合物結(jié)構(gòu)。結(jié)果 與模型組比較,CR-95E在6.25~50.00 μg· mL-1時可顯著抑制LPS誘導(dǎo)BV2細(xì)胞釋放NO(P<0.001),而CR-50E和CR-W無效,說明CR-95E具有確切的抗神經(jīng)炎作用。與模型組比較,CR-95E可顯著降低細(xì)胞上清液中炎癥因子IL-1β、IL-6和TNF-α的含量(P<0.05、0.01、0.001),顯著降低IDO的過量表達(dá)(P<0.001)。GC-MS分析,從CR-95E中共鑒定了34個化合物,主要為萜烯類和酮類成分,相對質(zhì)量分?jǐn)?shù)由高到低依次為異長葉烯酮(19.47%)、石竹烯氧化物(4.98%)、喇叭烯氧化物(-II)(4.01%)、α-古蕓烯(3.66%)等。結(jié)論 香附抗神經(jīng)炎癥活性部位為CR-95E,主要含有萜烯類和酮類成分等低極性成分,可通過抑制細(xì)胞NO和炎癥因子的釋放,抑制IDO過表達(dá)發(fā)揮抗神經(jīng)炎癥作用。
[Key word]
[Abstract]
Objective To evaluate the anti-neuroinflammatory activity and analyze its components of Cyperi rhizome. Methods 95% ethanol extract (CR-95E), 50% ethanol extract (CR-50E) and water extract (CR-W) of Cyperi rhizome were sequential prepared by heating reflux method. The anti-inflammatory activities were screened based on lipopolysaccharide (LPS) induced BV2 cells inflammation model. The content of nitric oxide (NO) and inflammatory factors (IL-1β、TNF-α、IL-6) in supernatant of model cells were detected after the obtained active site treatment. Indoleamine 2, 3-dioxygenase (IDO), the key enzyme of the kynurenine pathway rate-limiting enzyme, was detected by ELISA. The chemical constituents of the obtained active site was analyzed by GCMS, and further identified through searching by NIST MS search 2.0. Results CR-95E significantly decreased the content of NO (P < 0.001) in LPS-induced BV2 cells at 6.25-50.00 μg· mL-1, but CR-50E and CR-W did no, which suggested that the CR-95E had a definite anti-neuroinflammation effect. Moreover, CR-95E significantly decreased the contents of NO and intracellular inflammatory factors such as IL-1β, IL-6 and TNF-α. In addition, CR-95E could significantly reduce the overexpression of IDO, suggesting that CR-95E might play a role in cell protection and regulating neuronal dysfunction through inhibiting the accumulation of toxic metabolites caused by abnormal activation of kynurenine pathway. As a result, 34 compounds were identified by NIST MS Search 2.0, including isolongifolene (19.47%), caryophyllene oxide (4.98%), ledene oxide-(II) (4.01%), α-paleobrassene (3.66%). Conclusion The anti-neuroinflammatory components of Cyperi rhizome concentrate on low polarity components mainly including terpenes and ketones. Its plays an anti-neuroinflammation role through inhibiting the release of NO and inflammatory factor sby regulated the NF-κB inflammatory signaling pathway and kynurenine pathway. Our study provided scientific data for the discovery and mechanism of anti-neuroinflammatory components of Cyperi rhizome.
[中圖分類號]
R284
[基金項目]
國家自然科學(xué)基金資助項目(81973461);中國醫(yī)學(xué)科學(xué)院醫(yī)學(xué)與健康科技創(chuàng)新工程項目(2021-I2M-1-028)