以升麻为主的复方精油制备及其对小鼠的抗辐射作用

Preparation and analysis of compound essential oil based on cimicifuga and its antiradiation activity in mice

摘要:

以升麻、金银花、沙棘、黄芩和玉竹5种本草植物中药材为原材料，采用微波辅助提取法制备复方精油，采用单因素实验和响应面实验对提取工艺条件进行优化；通过GC-MS法对复方精油成分进行定性和定量分析；采用抑菌圈实验对复方精油的金黄色葡萄球菌抑菌活性进行检测；通过UVB照射法建立小鼠辐射皮肤损伤模型和人类永生化角质形成细胞（HaCaT）细胞的损伤模型；基于H-E染色法对超氧化物歧化酶（SOD）的活性、丙二醇（MDA）水平、胶原蛋白含量进行检测；通过Western blotting法测定HaCaT细胞p53和增殖细胞核抗原（PCNA）蛋白表达水平；探究复方精油对经UVB照射后的小鼠皮肤的辐射修复活性。结果表明，以体积分数75%的乙醇水溶液为提取溶剂，在液料比（mL∶g）为28∶1，微波功率为385 W，提取时间为32 min的最佳条件下，复方精油提取率为0.624%；复方精油中共检测出32种化学成分，包括醇类、酮类等，其中甲基丁香酚、3,4-二甲氧基苯乙烯、3,4,5-三甲氧基甲苯的相对含量分别为18.64%、13.27%、12.16%；8 μL复方精油对金黄色葡萄球菌的抑菌圈直径为(18.23)±(0.25) mm，抑菌属于高度敏感，其抑制效果主要源自黄芩精油；复方精油通过升高小鼠皮肤组织SOD活性和胶原蛋白的含量，降低小鼠皮肤组织丙二醛（MDA）水平，从而发挥对抗UVB辐射的预防作用，并可通过下调HaCaT细胞p53和PCNA的蛋白表达，从而对UVB辐射损伤起到修复作用。

Abstract:

The compound essential oil was prepared by microwave extraction method, and the preparation process was optimized by response surface methodology, qualitative and relative quantitative analysis of the compound essential oil was carried out by GC-MS. The anti UVB radiation activity of HaCaT cells was studied by establishing a mouse UVB radiation skin injury model and measuring the expression levels of p53 and PCNA proteins by Western blotting. The results showed that using a 75% ethanol aqueous solution as the extraction solvent, at a liquid to material ratio (mL: g) of 28:1, a power of 385 W, and an extraction time of 32 min, The extraction rate of compound essential oil is 0.624%; A total of 32 chemical components were detected in the compound essential oil, including alcohols, ketones, etc. The relative contents of methyl eugenol, 3,4-dimethoxystyrene, and 3,4,5-trimethoxytoluene were 18.64%, 13.27%, and 12.16%, respectively;The diameter of the inhibition zone of 8μL compound essential oil against Staphylococcus aureus is (18.23) ± (0.25) mm, indicating high sensitivity to inhibition. Its inhibitory effect mainly comes from Scutellaria baicalensis essential oil; Compound essential oils exert a preventive effect against UVB radiation by increasing the activity of SOD and the content of collagen in mouse skin tissue, reducing the level of malondialdehyde (MDA) in mouse skin tissue, and can also repair UVB radiation damage by downregulating the protein expression of p53 and PCNA in HaCaT cells.