川芎挥发油对HaCaT细胞和皮肤的刺激性研究

Stimulating effect of essential oils from Ligusticum chuanxiong Hort on HaCaT cells and skin

目的阐明川芎挥发油的皮肤刺激性。方法体外培养HaCaT细胞,分别采用四甲基偶氮唑盐比色法(MTT法)和酶联免疫吸附法(ELISA法),以经典化学促透剂油酸为对照,考察不同浓度川芎挥发油对皮肤HaCaT细胞活力、HaCaT细胞上清液中前列腺素E_2(PGE_2)含量的影响;采用豚鼠皮肤刺激评分表法考察不同浓度挥发油的皮肤累积刺激性,光学显微镜下观察豚鼠皮肤组织学的形态学改变。结果 0.005%、0.015%、0.025%挥发油组HaCaT细胞活力分别为0.005%油酸组的1.79、1.65、1.48倍,而0.05%挥发油组与0.005%油酸组差异无统计学意义。0.005%、0.015%、0.025%、0.05%挥发油对HaCaT细胞上清液的PGE_2含量影响分别为(0.99±0.08)%、(1.63±0.09)%、(0.98±0.09)%、(0.04±0.01)%,均低于0.005%油酸组(4.23±0.68)%(P<0.05)。各浓度挥发油豚鼠连续给药7 d,均仅见轻微红斑,未见水肿、皮肤隆起等,15%挥发油组红斑强度低于5%油酸组。挥发油仅在15%浓度时才出现角质层结构的改变,而5%油酸角质层就出现大面积剥离和缺失。结论川芎挥发油的皮肤刺激性低于油酸,有可能成为一种优良的化学促透剂。

Objective To investigate the skin stimulation effect of essential oils from Ligusticum chuanxiong Hort. Methods HaCaT cells were cultured in vitro. Using MTT and ELISA methods, we examined the effect of the essential oil （different concentrations） on HaCaT cell proliferation and prostaglandin E2 （PGE2） levels in culture supernatants of HaCaT cells, and the results were compared with those of oleic acid, a classic permeation enhancer. The cumulative skin stimulation effect was determined by visual scoring in guinea pigs and the histological changes were determined by light microscopy. Results The HaCaT cell viabilities of the 0. 005%, 0. 015%, and 0. 025% essential oil groups were 1.79-, 1.65-, and 1.48-fold that of the 0. 005% oleic acid group, respectively, and there was no significant difference between the 0.05% essential oil group and 0. 005% oleic acid group. The 0. 005%, 0. 015%, 0. 025%, and 0.05% essential oil influenced the supernatant PGE2 levels by （0.99±0.08） %, （ 1.63 ±0.09 ）%, （0.98 ± 0.09 ） %, and （ 0.04 ± 0.01 ） %, respectively, which were all significantly lower than the influence of 0. 005% oleic acid （[4. 23±0. 682 %, P〈 0.05）. Only slight erythema was observed after continuous administration the essential oil （different concentrations） for 7 days, with no edema or skin uplift, and the erythema caused by 15% essential oil was lower than that caused by 5% oleic acid. Only 15% essential oil exhibited the mechanical injury of the stratum corneum. And 5% oleic acid group showed stripped and lost stratum corneum over large areas.Conclusion Chuanxiong oil is less cytotoxic and less stimulating to the skin compared with oleic acid, and may become an excellent skin permeation enhancer.