薄层色谱生物自显影法筛选新疆莫合烟天然乙酰胆碱酯酶抑制剂的活性成分

TLC Bioautographic Method for Activity Screening of Natural Acetyl Cholinesterase Inhibitors from Xinjiang Mohe- Tobacco

目的为阐明烟草中的烟碱降低退行性疾病发病率的作用机理,对新疆特有的莫合烟进行天然乙酰胆碱酯酶抑制剂的活性筛选。方法采用薄层色谱分离和生物活性测定相结合的薄层色谱生物自显影技术,对新疆莫合烟提取液进行活性筛选。结果新疆莫合烟提取液中的烟碱通过薄层色谱生物自显影,在紫色背景下出现白色斑点,表明具有活性;但白色斑点较弱,表明提取液中的烟碱含量无法达到显著的抑制浓度,较对照品石杉碱甲活性偏低;2,2-二苯基苦基苯肼自由基(DPPH)显色法表明新疆莫合烟提取液同时具有较好的抗氧化活性。结论新疆莫合烟中的烟碱可与乙酰胆碱酯酶结合并抑制其活性。薄层色谱生物自显影方法用于天然乙酰胆碱酯酶抑制剂的活性筛选,操作简单,灵敏度和专属性高。

Objective To explain nicotine in tobacco can reduce the incidence of degenerative diseases,perform the activity screening of acetyl cholinesterase inhibitors of Xinjiang＇s unique Mohe-tobacco for illuminating the action mechanism. Methods The thin layer chromatography（TLC）bioautographic method combining TLC with biological activity assay was used to conduct the activity screening of extract solution of Xinjiang Mohe-tobacco. Results Nicotine in the extract solution of Xinjiang Mohe-tobacco appeared the white spots on the purple background by the TLC bioautography,indicating that which possesses the activity,but the white spots were weak, indicating that the nicotine content in the extract solution was unable to reach the significant inhibiting concentration,its activity was lower than that of the reference substance huperzine A;the DPPH development method revealed that the extraction solution of Xinjiang Mohe-tobacco simultaneously possesses the better antioxidant activity. Conclusion Nicotine in Xinjiang Mohe-tobacco can be com-bined with acetyl cholinesterase and inhibits its activity. The TLC bioautographic method is easy to operate with high sensitivity and specificity in the activity screening for natural acetyl cholinesterase inhibitor.