目的 观察豚鼠经皮给予沉香精油后的过敏反应,评价沉香精油的安全性。方法 选取健康雌性豚鼠30只,随机分为对照组、二甲苯组,水蒸气蒸馏提取和CO₂超临界萃取的沉香精油0.025、0.050mL组,每组5只。对照组给予蒸馏水0.05mL/只;二甲苯组给予二甲苯0.05mL/只;水蒸气蒸馏提取、CO₂超临界萃取沉香精油组分别给予0.025、0.050mL/只,各组均经皮给药。观察各组豚鼠皮肤过敏反应情况并拍照记录。摘取豚鼠脾脏并称质量,计算脾脏指数。酶联免疫法检测豚鼠血清和脾脏组织匀浆上清中免疫球蛋白(Ig)A、IgG、IgE的水平。取致敏皮肤组织,显微镜下观察病理学改变。结果 与对照组相比,沉香精油组给药1h后开始出现红斑、水肿等反应,4h时反应最明显,给药24h后开始逐渐消退,60~72h后基本恢复正常。沉香精油组豚鼠过敏期间出现明显的红肿以及严重的红斑的致敏率为25%~75%,其中CO₂超临界萃取的沉香精油过敏反应更显著,为中重度过敏。与对照组相比,水蒸气蒸馏提取和CO₂超临界萃取的沉香精油0.025、0.050mL组均使豚鼠脾脏指数水平显著降低(P<0.01)。与对照组相比,水蒸气蒸馏提取和CO₂超临界萃取的沉香精油0.025、0.050mL组对豚鼠血清中IgA、IgG、IgE水平没有显著性的影响,而在豚鼠脾脏组织匀浆上清中IgA、IgG、IgE水平均有不同程度升高(P<0.05、0.01)。病理学结果显示,与对照组相比,CO₂超临界萃取精油的皮肤过敏反应比较严重,而水蒸气蒸馏提取的沉香精油则有轻度炎症细胞浸润等现象。结论 沉香精油经皮肤涂抹给药对豚鼠有致敏作用,其中CO₂超临界萃取沉香精油的过敏反应更显著,这为沉香精油用药安全及用于日化等相关产品研制提供参考依据。

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

国家自然科学基金资助项目(82204657);国家重点研发计划项目(2018YFC1706400);中国医学科学院医学与健康科技创新工程(2021-I2M-1-032);海南省重点研发计划项目(ZDYF2022SHFZ030)