石菖蒲抑酶活性的实验研究

Research on Inhibitory Effect of Acorus tatarinouii Schott on AChE Activity in Vitro

目的建立体外抑制乙酰胆碱酯酶(Acetylcholinesterase,AchE)活性实验方法,探讨石菖蒲抑酶活性及活性部位。方法提取制备石菖蒲根和叶的乙醚、醋酸乙酯及正丁醇萃提液,以石油醚-醋酸乙酯(85∶15)为展开剂,对石菖蒲根和叶的乙醚萃提液层析分离后进行抑制AChE活性试验,采用分光光度法分别测定石菖蒲根和叶醋酸乙酯及正丁醇萃提液的AChE抑制率。结果石菖蒲根和叶的挥发油在薄层板显示有显著抑酶斑点,石菖蒲根和叶的乙醚萃提液薄层色谱中与α-细辛醚和细辛醚对应的显著抑酶斑点比移值为Rf0和Rf(0.64±0.01)[t=(17±2)℃];石菖蒲根和叶的醋酸乙酯及正丁醇萃提液酶抑制率分别为39.17%,18.45%,24.33%和1.74%;石菖蒲根的水溶液无明显抑酶斑点但测定仍有较弱酶抑制率,石菖蒲叶的水溶液无抑酶斑点且两者均显示蓝色斑点。结论石菖蒲根和叶的挥发油具较强抑制AChE活性,其中α-细辛醚为强抑酶活性成分,石菖蒲根醋酸乙酯和正丁醇萃提液抑酶活性分别较石菖蒲叶强,石菖蒲根水溶液有微弱抑酶活性而叶的水溶液无抑酶活性,但两者的水溶液均有酶促作用的成分。该法作为一种体外抑酶(AchE)活性实验,结果与文献报道的体内抑酶实验一致。

Objective To investigate the inhititory effect of Acorus tatarinouii Schott and its active part. Methods The solutions of roots and leaves were prepared from Acorus tatarinouii Schott with ether, acetic ether and n - butyric alcohol. The experiments of AchE inhibition was done by TLC for ether solution of root and leaf, the solvent system was petroleum ether - acetic ether（ 85： 15 ）. Determination of the AchE inhibition percentage was established for acetic ether and n - butyric alcohol solution of root and leaf by spectrophotometry. Results The volatile oil of root and leaf from Acorus tatarinouii Schott was marked white spot on TLC. There was marked white spots on TLC in ether solution of root and leaf, which was corresponding with α - asarone and reference volume were 0,0.64 ± 0.01 （ t = 17 ± 2℃ ）. The AchE inhibition percentage of effective parts in root and leaf from Acorus tathrinouii Schott after extraction by acetic ether and n -butyric alcohol were 39.17% ,18.45% ,24. 33% and 1.74%. There was no marked white spot on TLC in water solution of the root and leaf but rootg AchE inhibition percentage was 1.20% . There was blue spot. Conclusion The volatile oil of root and leaf from Acorus tatarinouii Schott has strong inhibvtiry effect on AchE activity and α - asarone was strong active substance. The activity of Ache Inhibtion of root extraction by acetic ether and n - butyric alcohol was stronger than leaf. The water solution of the root has weaker activity of AchE inhibition and the leaf has not. Both root and leaf have active substances of AchE promoter in their water solution. A new way to test the AchE inhibition in vitro is established and it reaches a consistent result in vivo.