目的 探究廣陳皮揮發(fā)油（GCPVO）調(diào)節(jié)Toll樣受體4（TLR4）/髓樣分化蛋白88（MyD88）/NOD樣受體熱蛋白結(jié)構(gòu)域相關(guān)蛋白3（NLRP3）/Caspase-1信號(hào)通路減輕脂多糖（LPS）誘導(dǎo)的肺上皮細(xì)胞炎癥作用及物質(zhì)基礎(chǔ)。方法 以不同陳化年份（1、3、5、7、9、15年）廣陳皮為研究對(duì)象，以電子鼻和氣相色譜-質(zhì)譜法（GC-MS）分別分析鑒別廣陳皮粉末、GCPVO的化學(xué)成分及不同陳化年份成分組成特征。建立LPS致BEAS-2B細(xì)胞炎癥模型，以細(xì)胞中丙二醛（MDA）、超氧化物歧化酶（SOD）水平，培養(yǎng)液中白細(xì)胞介素-1β（IL-1β）蛋白水平為檢測(cè)指標(biāo)，評(píng)價(jià)揮發(fā)油對(duì)肺上皮細(xì)胞炎癥的改善作用；采用Western blotting法檢測(cè)BEAS-2B細(xì)胞中NLRP3、Caspase-1、TLR4和MyD88蛋白的表達(dá)。以藥效學(xué)數(shù)據(jù)作為母序列，并以GC-MS分析得到不同陳化年份共有成分含量為特征序列，開展譜效關(guān)系研究，發(fā)現(xiàn)與活性相關(guān)的物質(zhì)。結(jié)果 電子鼻結(jié)果表明，廣陳皮含有更多的硫化合物、甲烷等短鏈烴類，萜烯，芳香化合物，樣品之間的距離相對(duì)較遠(yuǎn)，表明存在顯著差異；6個(gè)陳化年份GCPVO主要共有成分12種，在陳化時(shí)間1～9年中β-石竹烯的含量隨著年份的增加而降低，而α-側(cè)柏烯、α-蒎烯、萜品烯、月桂烯、D-檸檬烯、松油烯、4-蒈烯、2-（甲氨基）苯甲酸甲酯在陳化3年廣陳皮中相對(duì)含量最高。與模型組比較，GCPVO顯著降低LPS誘導(dǎo)的IL-1β和MDA含量顯著升高，提高SOD活性，顯著降低LPS誘導(dǎo)的BEAS-2B細(xì)胞中炎癥相關(guān)蛋白TLR4、MyD88、NLRP3、Caspase-1的表達(dá)水平（P＜0.05、0.01），陳化3年樣品效果最好。GCPVO中的化合物D-檸檬烯、α-蒎烯、松油烯與改善肺上皮細(xì)胞炎癥的譜效相關(guān)性最高。結(jié)論 GCPVO調(diào)節(jié)TLR4/MyD88/NLRP3/Caspase-1信號(hào)通路減輕LPS誘導(dǎo)的肺上皮細(xì)胞炎癥，D-檸檬烯、α-蒎烯、松油烯是發(fā)揮藥效的關(guān)鍵物質(zhì)。

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.

中央本級(jí)重大增減支項(xiàng)目（2060302）；云浮市2021年中醫(yī)藥（南藥）產(chǎn)業(yè)人才項(xiàng)目-南藥資源創(chuàng)新團(tuán)隊(duì)