灵芝蜂蜜面膜功效及安全性评价研究

Evaluation of efficacy and safety for a facial mask made of ganoderma lucidum and honey

对灵芝蜂蜜面膜进行活性和安全性评价研究,为传统中药面部外用制剂的应用提供实验数据支持。将灵芝蜂蜜面膜用水、50%乙醇以及90%乙醇进行提取,制备不同溶剂提取液。将不同溶剂制备的灵芝蜂蜜面膜提取液进行DPPH自由基清除实验,研究不同溶剂提取液的抗氧化作用;将灵芝蜂蜜面膜水、90%乙醇提取液作用于人永生化角质形成细胞(HaCaT细胞),用MTT法筛选出安全给药浓度;采用RT-qPCR法检测灵芝蜂蜜面膜对HaCaT细胞中基质金属蛋白酶-1(MMP-1)和基质金属蛋白酶-9(MMP-9)的mRNA表达水平的影响;将灵芝蜂蜜面膜涂抹于昆明小鼠皮肤表面,进行动物急性皮肤毒性实验,评价其安全性。结果表明,不同溶剂制备的灵芝蜂蜜面膜提取液均能清除活性氧自由基,其中水提取液IC_(50)为14.123 mg/mL、50%乙醇提取液IC_(50)为22.651 mg/mL、90%乙醇提取液IC_(50)为22.140 mg/mL;灵芝蜂蜜面膜水提取液和90%乙醇提取液低剂量时可以增强HaCaT细胞的细胞活力,具有细胞保护作用。与模型组相比,处理组HaCaT细胞中MMP-1和MMP-9的mRNA表达水平明显降低。动物实验结果表明:灵芝蜂蜜面膜对昆明小鼠皮肤无毒性。灵芝蜂蜜面膜具有良好的抗氧化作用,可以有效抑制细胞外基质的降解、延缓细胞的衰老,对皮肤无毒性,具有高度安全性。

The efficacy and safety of a facial mask made of ganoderma lucidum and honey were evaluated to provide experimental data support for the application of traditional Chinese medicine mask.The facial mask made of ganoderma lucidum and honey was extracted with water,50%ethanol and 90%ethanol to prepare its extracts by different solvents.In vitro DPPH free radical scavenging tests were carried out to study the function of antioxidation for the extracts from the facial mask made of ganoderma lucidum and honey with different solvents.The water extract and 90%-ethanol extract were applied to HaCaT cells,and the safe concentration was screened by MTT assay.RT-qPCR method was used to detect the effects of the facial mask on the mRNA expression levels of matrix metalloproteinase-1(MMP-1)and matrix metalloproteinase-9(MMP-9)in HaCaT cells.The facial mask was applied to the skin surface of KM mice,and thus the skin acute toxicity test in animals was used to evaluate its safety.The results show that all the extracts made with different solvents from the facial mask can scavenge reactive oxygen species,in which the water extract shows an IC_(50) of 14.123 mg·mL^(-1),the 50%-ethanol extract shows and IC_(50) of 22.651 mg·mL^(-1) and the 90%-ethanol extract shows an IC_(50) of 22.140 mg·mL^(-1);the water extract and 90%-ethanol extract from the facial mask made of ganoderma lucidum and honey can enhance the viability of HaCaT cells and play the role of protecting cells at low dosage.Compared with the model group,the mRNA expression levels of MMP-1 and MMP-9 in HaCaT cells of the treatment group are significantly decreased.Animal experimental results show that,the facial mask made of ganoderma lucidum and honey has no toxicity for KM mice skin.Therefore,the facial mask made of ganoderma lucidum and honey has good antioxidant effect,can effectively inhibit the degradation of extracellular matrix,delay cell senescence,and it is nontoxic to skin and has a high degree of safety.