目的 探究广陈皮挥发油（GCPVO）调节Toll样受体4（TLR4）/髓样分化蛋白88（MyD88）/NOD样受体热蛋白结构域相关蛋白3（NLRP3）/Caspase-1信号通路减轻脂多糖（LPS）诱导的肺上皮细胞炎症作用及物质基础。方法 以不同陈化年份（1、3、5、7、9、15年）广陈皮为研究对象，以电子鼻和气相色谱-质谱法（GC-MS）分别分析鉴别广陈皮粉末、GCPVO的化学成分及不同陈化年份成分组成特征。建立LPS致BEAS-2B细胞炎症模型，以细胞中丙二醛（MDA）、超氧化物歧化酶（SOD）水平，培养液中白细胞介素-1β（IL-1β）蛋白水平为检测指标，评价挥发油对肺上皮细胞炎症的改善作用；采用Western blotting法检测BEAS-2B细胞中NLRP3、Caspase-1、TLR4和MyD88蛋白的表达。以药效学数据作为母序列，并以GC-MS分析得到不同陈化年份共有成分含量为特征序列，开展谱效关系研究，发现与活性相关的物质。结果 电子鼻结果表明，广陈皮含有更多的硫化合物、甲烷等短链烃类，萜烯，芳香化合物，样品之间的距离相对较远，表明存在显著差异；6个陈化年份GCPVO主要共有成分12种，在陈化时间1～9年中β-石竹烯的含量随着年份的增加而降低，而α-侧柏烯、α-蒎烯、萜品烯、月桂烯、D-柠檬烯、松油烯、4-蒈烯、2-（甲氨基）苯甲酸甲酯在陈化3年广陈皮中相对含量最高。与模型组比较，GCPVO显著降低LPS诱导的IL-1β和MDA含量显著升高，提高SOD活性，显著降低LPS诱导的BEAS-2B细胞中炎症相关蛋白TLR4、MyD88、NLRP3、Caspase-1的表达水平（P＜0.05、0.01），陈化3年样品效果最好。GCPVO中的化合物D-柠檬烯、α-蒎烯、松油烯与改善肺上皮细胞炎症的谱效相关性最高。结论 GCPVO调节TLR4/MyD88/NLRP3/Caspase-1信号通路减轻LPS诱导的肺上皮细胞炎症，D-柠檬烯、α-蒎烯、松油烯是发挥药效的关键物质。

Objective To explore the modulation of toll like receptor 4 (TLR4)/myeloid differentiation protein 88 (MyD88)/NOD like receptor thermal domain associated protein 3 (NLRP3)/Caspase-1 signaling pathway by Guangchenpi volatile oil (GCPVO) to alleviate lipopolysaccharide (LPS) - induced pulmonary epithelial cell inflammation and its material basis. Methods Using aged Guangchenpi from different years (1, 3, 5, 7, 9, 15) as the research object, electronic nose and gas chromatography-mass spectrometry (GC-MS) were used to analyze and identify the chemical composition of Guangchenpi powder and GCPVO, as well as the compositional characteristics of components from different aged years. The BEAS-2B cell model of inflammation was established induced by lipopolysaccharide (LPS). The disturbance of malondialdehyde (MDA), superoxide dismutase (SOD) and interleukin 1β (IL-1β) attributed with oxidative stress and inflammation have been evaluated. The expression of the proteins NLRP3, Caspase-1, TLR4 and MyD88 were also detected by Western blotting. Then the spectrum-effect relationship was analyzed by using original data of the content of common peaks and pharmacodynamics indicators. Results The results of the electronic nose showed that Guangchenpi contains more sulfur compounds, methane and other short-chain hydrocarbons, terpenes, aromatic compounds, and the samples are relatively distant from each other, indicating significant differences. The main common components of the 6 aged GCPVO were 12, and the content of β-caryophyllene decreased with the increase of aging time from 1 to 9 years, while the relative content of α-pinene, limonene, terpineol, myrcene, D-limonene, turpentine, 4-carene, and 2-(methylamino) benzaldehyde was the highest in the aged GCPVO samples aged 3 years. Compared with the model group, GCPVO significantly reduced the content of LPS-induced IL-1β and MDA, increased the activity of SOD, and significantly reduced the expression levels of inflammation-related proteins TLR4, MyD88, NLRP3, and Caspase-1 in LPS-induced BEAS-2B cells (P <0.05, 0.01), with the aged 3 years sample showing the best effect. The compounds D-limonene,α-pinene, and turpentine in GCPVO showed the highest correlation with the anti-inflammatory spectrum in lung epithelial cells. Conclusion GCPVO regulates the TLR4/MyD88/NLRP3/Caspase-1 signaling pathway to alleviate LPS-induced inflammation in lung epithelial cells, and D-limonene, α-pinene, and turpentine are the key substances for the drug effect.

中央本级重大增减支项目（2060302）；云浮市2021年中医药（南药）产业人才项目-南药资源创新团队