甘肃产艾叶挥发油的化学成分及遗传毒性研究

目的:分析甘肃产艾叶挥发油的主要化学成分,探讨其对小鼠的遗传毒性.方法:①成分分析:用气相色谱-质谱法分析艾叶挥发油的化学成分,用归一化法测定各组分的相对百分含量.②微核试验:选用昆明种孕鼠30只,随机分为5组:正常对照组,ig给予等容积花生油；艾叶油2,1,0.5 mL·kg-1组,ig给予艾叶油；阳性对照组,ip给予环磷酰胺(CP)40 mg·kg-1.正常对照组和艾叶油组自孕第12天开始ig给药,连续5d.阳性对照组于孕第15天ip给予CP,连续2d.各组孕鼠均于孕第16天用药后处死,取小鼠骨髓细胞和胎鼠肝脏做微核试验.③精子畸形试验:昆明种小鼠雄性40只,随机分为5组,各组用药剂量及给药方式同微核试验,均连续5d.首次给药后35 d处死,取出其附睾制片,读取精子畸形率.结果:从甘肃产艾叶挥发油分离出72个峰,其中鉴定了56个成分,占挥发油色谱峰面积的90.83％.艾叶挥发油灌胃剂量2 mL·kg-1时,孕鼠和雄鼠诱发的骨髓微核率、胚胎肝微核率和精于畸形率均较阴性对照组显著升高(P＜0.05).剂量为1 mL·kg-1时,诱发的胚胎肝微核率较阴性对照组显著升高(P＜0.05),骨髓微核率与精子畸形率与阴性对照组相比,无显著性差异.剂量为0.5mL·kg-时,诱发的骨髓微核率、胚胎肝微核率、精子畸形率与阴性对照组相比,均无显著性差异.结论:甘肃产艾叶挥发油的主要成分为桉叶素(20.45％)、蒿醇(12.12％)、樟脑(6.99％)、青蒿酮(5.85％)、左旋龙脑(4.27％)等；一定剂量的艾叶挥发油对小鼠具有潜在的遗传毒性,并呈剂量-反应关系.

Study on Chemical Constituents and Genotoxicity of Volatile Oil in Artemisia argyi from Gansu

Objective:To analyze the main chemical constituents of volatile oil in Artemisia argyi from Gansu,and explore its genotoxicity in mice.Method:①Constituents analysis: the chemical constituents of volatile oil in A.argyi were separated and identified by GC-MS,the relative content of each component was determined by area normalization.②Micronucleus assay: thirty Kunming pregnant mice were randomly divided into 5 groups: normal control group,ig given with peanut oil in equal volume;volatile oil 2,1,0.5 mL·kg-1 groups,ig given volatile oil;and positive control group,ip given cyclophosphamide(CP) 40 mg·kg-1.The normal control group and volatile group were ig administrated once a day from the 12th day of pregnancy to the 16 th day,for 5 d,while positive control group was ip given CP on the 15th day of pregnancy for 2 d.On the 16th day,the pregnant mice in each group were sacrificed to take out the embryo from mice,micronucleus test of adult mice bone marrow cells and fetal mice hepatocytes were carried out.③Sperm abnormality assay: forty male mice were randomly divided into 5 groups,the doses and administration of each group were similar to micronucleus test.The drugs was administered for 5 days.After 35 d of first medication,the mice were sacrificed to take out the epididymis and produce the sperm smear,and examination of sperm deformity was carried out.Result: Seventy-two components were isolated from volatile oil and fifty-six of them were identified,which was about 90.83% of all components.Compared with control group,the micronucleus rates of bone marrow and fetal hepatic cell and teratospermia number were all significantly increased(P<0.05) at a oral dose of 2 mL·kg-1;only the micronucleus rate on fetal liver was increased significantly(P<0.05) with the oral dose of 1 mL·kg-1;there was no statistic difference in any of the indexes at the dose of 0.5 mL·kg-1.Conclusion: The main chemical components of volatile oil in A.argyi from Gansu are 1,8-cineole(20.45%),artemisia alcohol(12.12%),camphor(6.99%),Artemisia ketone(5.85%),L-borneol(4.27%),et al.A potential dose-related genotoxicity of volatile oil in A.argyi has been verified in this research.