目的 沉香是常用名贵药材,因具有多种药效且药用资源稀缺,是目前研究的热点。沉香精油(Chinese eaglewood essential oil, CEEO)是其主要有效成分,本实验旨在研究CEEO的抗炎作用及分子机制。方法 采用小鼠耳肿胀实验和大鼠足肿胀实验评价CEEO的抗炎活性;通过脂多糖(LPS)诱导RAW264.7细胞模型研究CEEO对白细胞介素(IL)-1β、IL-6等的影响;采用Western blot方法研究CEEO对大鼠组织蛋白p-STAT3,p-IκBα/p-NF-κBp65的影响;通过气相色谱-质谱联用(GC-MS)方法分析CEEO的化学成分。结果 CEEO (60~960 mg·kg-1, ig)剂量依赖地抑制二甲苯所致耳肿胀(P<0.001),最高抑制率为73%;CEEO (680 mg·kg-1, ig)明显抑制角叉菜胶所致足肿胀(P<0.001),最高抑制率为51%。ELISA和RT-PCR结果显示,CEEO(10、5、2.5 μg·mL-1)显著抑制IL-1β和IL-6的产生及其mRNA水平(P<0.001),最高抑制率分别为68%和39%,64%和52%。Western blot结果显示,CEEO(680 mg·kg-1, ig)明显抑制p-STAT3蛋白的表达,抑制率为21% (P<0.05)。通过GC-MS鉴定了沉香中的117个化合物,其主要成分为倍半萜类(68.83%)。结论 本实验首次发现中国沉香精油灌胃给药具有显著的抗炎活性,其机制可能是通过降低p-STAT3的过表达,进而抑制前炎症细胞因子IL-1β和IL-6的产生和释放,从而抑制炎症的发生与发展。沉香精油的抗炎作用可能与其所含的主成分倍半萜有关。
Abstract
OBJECTIVE Eaglewood is a commonly used precious herbal medicine. Due to the multiple drug effects and shortage of medicinal resources, it is currently a hot research topic. Chinese agarwood essential oil (CEEO) is the main active ingredient of eaglewood. To investigate the anti-inflammatory effect and mechanism of CEEO. METHODS The anti-inflammatory effect of CEEO was evaluated on xylene-induced ear edema in mice and carrageenan-induced paw swelling in rats. The production and transcription levels of pro-inflammatory cytokines IL-1β and IL-6 were detected by ELISA or quantitative RT-PCR in LPS-activated RAW264.7 macrophages. The protein levels of p-transducer and activator of transcription 3 (STAT3) and p-IκBα/p-NF-κBp65 were detected by Western blot analysis. The chemical profiles of CEEO were identified by GC-MS. RESULTS Intragastric administration with CEEO (60-960 mg·kg-1 in mice and 680 mg·kg-1 in rats) produced significant anti-inflammatory effect in vivo, the highest inhibition percentages were 73% and 51%, respectively (P<0.001). After treatment with CEEO (10, 5, and 2.5 μg·mL-1), ELISA and RT-PCR analysis showed that the production and mRNA level of IL-1β and IL-6 were attenuated by 68% and 39%, 64% and 52% (P<0.001), respectively, at the highest concentration. Western blot analysis showed that CEEO (680 mg·kg-1, ig) significantly restrained the phosphorylation of STAT3 by 21% (P<0.05). Totally 117 compounds were identified in CEEO by GC-MS analysis and the chemical analysis revealed that sesquiterpenoids were the major compounds (68.83%). CONCLUSION This study shows for the first time that intragastric administration of CEEO has significant anti-inflammatory effect, which may be due to the suppression of pro-inflammatory cytokines IL-1β and IL-6 by reducing the overexpression of p-STAT3, thereby inhibiting the occurrence and development of inflammation. The anti-inflammatory effect of CEEO may be related to the primary component sesquiterpenoids.
关键词
沉香精油 /
抗炎作用 /
转录因子 /
转录激活因子3 (STAT3) /
白细胞介素-1β /
白细胞介素-6 /
倍半萜
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Key words
Chinese agarwood essential oil /
anti-inflammation /
transducer /
activator of transcription 3 /
interleukin-1β /
interleukin-6 /
sesquiterpenoid
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中图分类号：
R282
R965
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脚注
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基金
国家重点研发计划“中医药现代化研究”重点专项项目资助(2018YFC1706402)
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