目的 觀察豚鼠經(jīng)皮給予沉香精油后的過(guò)敏反應(yīng),評(píng)價(jià)沉香精油的安全性。方法 選取健康雌性豚鼠30只,隨機(jī)分為對(duì)照組、二甲苯組,水蒸氣蒸餾提取和CO₂超臨界萃取的沉香精油0.025、0.050mL組,每組5只。對(duì)照組給予蒸餾水0.05mL/只;二甲苯組給予二甲苯0.05mL/只;水蒸氣蒸餾提取、CO₂超臨界萃取沉香精油組分別給予0.025、0.050mL/只,各組均經(jīng)皮給藥。觀察各組豚鼠皮膚過(guò)敏反應(yīng)情況并拍照記錄。摘取豚鼠脾臟并稱(chēng)質(zhì)量,計(jì)算脾臟指數(shù)。酶聯(lián)免疫法檢測(cè)豚鼠血清和脾臟組織勻漿上清中免疫球蛋白(Ig)A、IgG、IgE的水平。取致敏皮膚組織,顯微鏡下觀察病理學(xué)改變。結(jié)果 與對(duì)照組相比,沉香精油組給藥1h后開(kāi)始出現(xiàn)紅斑、水腫等反應(yīng),4h時(shí)反應(yīng)最明顯,給藥24h后開(kāi)始逐漸消退,60~72h后基本恢復(fù)正常。沉香精油組豚鼠過(guò)敏期間出現(xiàn)明顯的紅腫以及嚴(yán)重的紅斑的致敏率為25%~75%,其中CO₂超臨界萃取的沉香精油過(guò)敏反應(yīng)更顯著,為中重度過(guò)敏。與對(duì)照組相比,水蒸氣蒸餾提取和CO₂超臨界萃取的沉香精油0.025、0.050mL組均使豚鼠脾臟指數(shù)水平顯著降低(P<0.01)。與對(duì)照組相比,水蒸氣蒸餾提取和CO₂超臨界萃取的沉香精油0.025、0.050mL組對(duì)豚鼠血清中IgA、IgG、IgE水平?jīng)]有顯著性的影響,而在豚鼠脾臟組織勻漿上清中IgA、IgG、IgE水平均有不同程度升高(P<0.05、0.01)。病理學(xué)結(jié)果顯示,與對(duì)照組相比,CO₂超臨界萃取精油的皮膚過(guò)敏反應(yīng)比較嚴(yán)重,而水蒸氣蒸餾提取的沉香精油則有輕度炎癥細(xì)胞浸潤(rùn)等現(xiàn)象。結(jié)論 沉香精油經(jīng)皮膚涂抹給藥對(duì)豚鼠有致敏作用,其中CO₂超臨界萃取沉香精油的過(guò)敏反應(yīng)更顯著,這為沉香精油用藥安全及用于日化等相關(guān)產(chǎn)品研制提供參考依據(jù)。

Objective To evaluate the safety of agarwood essential oil by observing the allergic reaction of agarwood essential oil exposed to guinea pig skin. Methods Thirty healthy female guinea pigs were randomly divided into control group, dimethyl benzene group, and the 0.025 and 0.050 mL groups of steam distillation and CO₂ supercritical extraction agarwood essential oils, each group had 5 guinea pigs. Cotrol group was given distilled water 0.05 mL, dimethyl benzene group was given dimethyl benzene 0.05 mL, the agarwood essential oil group were given 0.025 and 0.050 mL agarwood essential oil extracted by steam distillation and extracted by CO₂ supercritical extraction, all groups were given percutaneous administration. The skin allergic reaction of guinea pigs was observed and photographed. The spleen of guinea pig was extracted and weighed to calculate the spleen index. The levels of IgA, IgG, and IgE in the supernatant of serum and spleen tissue were determined by enzyme-linked immunoassay. The pathological changes of sensitized skin tissues were observed under microscope. Results Compared with the control group, the reaction of erythema and edema began to appear in the agarwood essential oil group 1 h after administration, and the reaction was the most obvious at 4 h. The reaction gradually subsided after 24 h, and basically returned to normal after 60-72 h. The sensitization rate of obvious redness and severe erythema was 25%-75% in the agarwood essential oil group, among which the agarwood essential oil with CO₂ supercritical extraction had a more significant allergic reaction, which was moderate to severe allergy. Compared with the control group, water vapor extraction and CO₂ supercritical extraction of agarwood essential oil in 0.025 and 0.050 mL groups significantly reduced the spleen index of guinea pigs (P＜0.01). Compared with the control group, IgA, IgG, and IgE levels in serum of guinea pigs in 0.025 and 0.050 mL groups of agarwood essential oil extracted by steam distillation and CO₂ supercritical extraction had no significant effects. The levels of IgA, IgG, and IgE in the homogenate supernatant of spleen tissue of guinea pigs were increased in different degrees (P＜0.05, 0.01). The pathological results showed that compared with the control group, the skin allergic reaction of the essential oil extracted by CO₂ supercritical fluid extraction was more serious, while the essential oil extracted by steam distillation had mild inflammatory cell infiltration and other phenomena. Conclusion Agarwood essential oil could induce the allergic reaction, which more significant of CO₂ supercritical extraction, and provided reference and guidance for the safety of agarwood essential oil and the development of daily chemical products.

R285.5

國(guó)家自然科學(xué)基金資助項(xiàng)目(82204657);國(guó)家重點(diǎn)研發(fā)計(jì)劃項(xiàng)目(2018YFC1706400);中國(guó)醫(yī)學(xué)科學(xué)院醫(yī)學(xué)與健康科技創(chuàng)新工程(2021-I2M-1-032);海南省重點(diǎn)研發(fā)計(jì)劃項(xiàng)目(ZDYF2022SHFZ030)