瑶药长叶铁角蕨挥发油萃取工艺优化及其抗炎镇痛活性研究

Extraction the Volatile Oil from Yao Medicine Asplenium prolongatum Hook. and Study on Its Antiinflammatory and Analgesic Activities

目的研究瑶药长叶铁角蕨挥发油萃取工艺及其抗炎镇痛活性。方法采用单因素试验和正交试验,以挥发油萃取率为评价指标,考察萃取温度、萃取压力、萃取时间三个考察因素的最佳水平;采用二甲苯致小鼠耳肿胀法和角叉菜胶致小鼠足肿胀法观察其抗炎作用,采用热板致痛法和小鼠扭体法观察其镇痛作用,用酶联免疫吸附法(Elisa)法测定血清中肿瘤坏死因子α(TNF-α)和白细胞介素-1β(IL-1β)的含量,紫外分光光度法检测血清和炎性组织中前列腺素E2(PGE2)的含量。结果优化挥发油最佳萃取工艺为萃取温度40℃,萃取压力16 MPa,萃取时间150 min;长叶铁角蕨挥发油高、中、低剂量组(0.6 g·kg-1、0.3 g·kg-1、0.15 g·kg-1)对小鼠耳肿胀、足肿胀、热板致痛、扭体反应均具有抑制作用,其中高剂量组对耳廓肿胀和扭体反应的抑制率分别为60.9%、43.7%,各组与空白组相比较均具有显著性差异(P <0.05, P <0.01),均可不同程度降低小鼠血清TNF-α、IL-1β,小鼠血清及炎性组织中PGE2的含量。长叶铁角蕨挥发油高剂量组可降低小鼠腹腔毛细血管通透性,表现为OD值明显降低及明显降低CMC-Na致小鼠腹腔白细胞游走。结论优选的长叶铁角蕨挥发油萃取工艺萃取率高、重复性好,长叶铁角蕨具有显著的抗炎镇痛活性。

Objective To optimize the extraction process of volatile oil from Yao medicine Asplenium prolongatum Hook.and observe its anti-inflammatory and analgesic activities.Methods Using the extraction rates of volatile oil as index,the single factor experiment and orthogonal experimental design were employed to optimize three extraction factors:temperature,pressure and extraction duration.The models of ear edema induced by xylene and paw edema induced by carrageenan in mice were used to evaluate the anti-inflammatory activity.The hot-plate test and writhing assay in mice were used to evaluate the analgesic activity.Contents of TNF-α,IL-1β,PGE2 were measured by ELISA and radio-immunity method.Results The optimum process condition were extracting with a temperature of40℃,an extraction pressure of 16 MPa and an extraction duration of 150 min.High,middle and low dose groups(0.6 g·kg-1,0.3 g·kg-1,0.15 g·kg-1)of A.prolongatum volatile oil showed inhibition effect on 4 types of models.The inhibition rates of high dose group on the model of ear edema induced by xylene and writhing assay in mice were60.9%and 43.7%.Compared with the blank group,the inhibition rates showed significantly difference(P<0.05,P<0.01).The contents of PGE2 in serum of the mice’s inflammatory tissue,the contents of TNF-α,IL-1βin serum of the mice inflammatory induced by carrageenan were decreased.High dose of A.prolongatum also decreased the abdominal capillary permeability in mice.Conclusion The extraction process showed a high yield and good reproducibility.The A.prolongatum volatile oil possesses significant anti-inflammatory and analgesic activities.