罗仙子油精制工艺优化及其对D-半乳糖致小鼠皮肤衰老的作用研究

Optimization of the refining technology of maggots oil and its effect on skin aging induced by D-galactose in mice

目的:优化罗仙子油的精炼工艺,研究罗仙子精油对D-半乳糖致小鼠皮肤衰老的干预作用。方法:分别考察不同因素对罗仙子油的水化脱胶、碱炼脱酸精炼效果的影响,分析比较罗仙子油精制前后的理化值。将昆明种雌性小鼠随机分为正常组、模型组、基质组、维生素E组、罗仙子精油低剂量组和高剂量组。除正常组外,其余各组均每日颈背部皮下注射3%D-半乳糖连续42 d,建立小鼠亚急性衰老模型;各给药组于小鼠颈背部涂抹相应药物qd,持续32 d,HE染色观察皮肤组织形态,检测皮肤含水量、超氧化物歧化酶(SOD)活性、谷胱甘肽过氧化物酶(GSH-Px)活力、羟脯氨酸(HYP)及丙二醛(MDA)的含量。采用家兔对罗仙子精油进行皮肤刺激性实验,将12只家兔分为正常组、完整皮肤组、破损皮肤组。在家兔背部左侧给予0.5 m L罗仙子精油,右侧给予同体积基质,qd,连续7 d。观察药物对家兔皮肤的刺激性,对皮肤红斑和水肿进行评分,于末次给药后72 h对给药部位皮肤进行HE染色,并观察皮肤组织形态。结果:罗仙子油精炼后,碘值(18.714±0.116) g·100 g^(-1)、酸值(0.465±0.006) mg·g^(-1)、过氧化值(0.016±0.001) g·100 g^(-1)和皂化值(130.212±1.623) mg·g^(-1),符合按摩精油及润肤油的国家标准。与模型组比较,罗仙子精油组小鼠皮肤真皮层增厚,毛囊数明显增加且胶原纤维增多,皮肤含水量、HYP含量、SOD及GSH-PX活力明显提高(P<0.05,P<0.01),MDA含量显著降低(P<0.01)。在家兔皮肤刺激实验中,各组家兔的红斑和水肿评分均为0分,皮肤刺激性均评价为无刺激。结论:罗仙子精油具有延缓D-半乳糖致小鼠皮肤衰老的作用,且无刺激性,具有潜在应用价值。精炼后罗仙子油的色泽、气味、澄明度和流动性均得到改善,为罗仙子精油的抗衰老产品开发提供依据。

Objective:To optimize the refining technology of maggots oil and study the effect of refined maggots oil on skin aging induced by D-galactose in mice.Methods:The effects of different factors on the refining technology (including hydration degumming and deacidification by alkali-refining) of maggots oil were investigated,and the physical and chemical properties of maggots oil were analyzed.Female Kunming mice were randomly divided into normal group,model group,matrix group,vitamin E group,low dose and high dose refined maggots oil groups.Except for the normal group,the mice in the other groups were subcutaneously injected 3%D-galactose into the back of the neck for 42 consecutive days to establish the subacute aging model.The mice in each administration group were smeared with corresponding drugs once a day for 32 days.HE staining was used to observe the micromorphology of skin tissue,and the contents of water,superoxide dismutase (SOD),glutathione peroxidase(GSH-Px),hydroxyproline (HYP) and malondialdehyde (MDA) in the skin were detected.Ten rabbits were divided into normal group,intact skin group and damaged skin group.The skin irritation test was conducted on rabbits.Twelve rabbits were divided into normal group,intact skin group and damaged skin group.The rabbits were given 0.5 m L maggots essential oil on the left side of the back and olive oil of the same volume on the right side,once a day for 7 days.The irritation effect of the drug on the rabbit skin was observed,and the skin erythema and edema were scored.The skin of the administration site was stained with HE at 72 h after the last administration,and the skin tissue morphology was observed.Results:After refining,the iodine value,acid value,peroxide value and saponification value of maggots oil were (18.714±0.116) g·100 g^(-1),(0.465±0.006) mg·g^(-1),(0.016±0.001) g·100 g^(-1)and (130.212±1.623) mg·g^(-1),respectively,which conformed to the national standard of the massage essential oil and emollient oil.Compared with the model group,the dermis of mice in the maggots essential oil groups were thickened,the numbers of hair follicles and collagen fibers,the contents of water,HYP,SOD and GSH-Px in the skin increased significantly (P<0.05,P<0.01),the content of MDA decreased significantly(P<0.01).In the rabbit skin irritation test,the erythema and edema score of each group were 0,and the skin irritation was evaluated as no irritation.Conclusion:Maggots essential oil has anti-aging effect on mouse skin induced by D-galactose without irritation,and has potential application value.The color,smell,clarity and fluidity of maggots essential oil are improved after refining,which provides the basis for the development of anti-aging products of maggots essential oil.