壮药白花九里明提取物中咖啡酸含量测定及对子宫平滑肌作用

Determination of caffeic acid in Zhuang medicine Blumea megacephala (Randeria) Chang et Tseng extract and its effect on uterine smooth muscle

目的:测定壮药白花九里明提取物(下称提取物A)咖啡酸的含量,并考察其对小鼠离体子宫平滑肌收缩的影响。方法:采用高效液相色谱法测定提取物A咖啡酸含量;选用健康未孕雌性成年小鼠,随机分15组,每组10只,即生理盐水组、缩宫素组、8种不同浓度九里明提取物A组、0.32 mg/ml提取物A联合用缩宫素、阿托品、心得安、乙酰胆碱组。结果:九里明提取物A中含有咖啡酸,咖啡酸在1.2μg/ml^1.95μg/ml范围内与峰面积呈良好线性关系,回归方程Y=12269X+150.99,R^2=0.9996。加样平均回收率99.75%,RSD为0.4%,提取物A测量得咖啡酸含量平均值为112.3μg/g。与生理盐水组比较,浓度在0.16 mg/ml^0.64 mg/ml九里明提取物A收缩活力、收缩频率和平均张力有明显变化,呈兴奋作用。提取物A随着浓度增大收缩活力明显增强,M受体阻断剂阿托品与0.32 mg/ml提取物A联用收缩活力和频率均明显小于单独0.32 mg/ml提取物A。β1和β2肾上腺素受体阻滞剂心得安与0.32 mg/ml提取物A联用收缩活力和频率均远小于单独0.32 mg/ml提取物A。M、N受体兴奋药乙酰胆碱与0.32 mg/ml提取物A联用收缩活力和频率均远大于单独0.32 mg/ml提取物A。结论:九里明提取物A中含具有止血作用的咖啡酸,含量平均值为112.3μg/g。提取物A能兴奋子宫平滑肌,可能是通过影响M受体或N受体,与非选择性的β受体而实现的。

Objective:The content of caffeic acid in Blumea megacephala(Randeria)Chang et Tseng extract(hereinafter referred to as extract A) was determined, and its effect on the isolated uterine smooth muscle of mice was examined. Methods:High-performance liquid chromatography was used to determine caffeic acid content of extract A. Healthy adult female mice were randomly divided into 15 groups, 10 in each group, saline group, oxytocin group, 8 different concentrations of extract A groups and extract A combined with oxytocin, atropine, propranolol, and acetylcholine groups. Results: Extract A contained caffeic acid, caffeic acid had a good linear relationship with peak area in the range of 1.2μg/ml^1.95μg/ml, the regression equation is y=12269 x+150.99, R^2=0.9996, the average recovery rate was 99.75%, and RSD was 0.4%. The average content value of caffeic acid in extract A was 112.3μg/g. Compared with saline group, the contraction activity, shrinkage frequency and average tension showed significant changes in 0.16 mg/ml^0.64 mg/ml groups. The contraction activity of extract A significantly increased according to the increasing concentration. The contraction activity and frequency of the M receptor blocker atropine combined with 0.32 mg/ml extract A were significantly lower than that of 0.32 mg/ml extract A alone. The contractile activity and frequency of β1 and β2 adrenergic receptor blocker propranolol combined with 0.32 mg/ml of Extract A were significantly lower than those of 0.32 mg/ml Extract A alone. The contractile viability and frequency of M and N receptor stimulant acetylcholine combined with 0.32 mg/ml of extract A were significantly higher than those of 0.32 mg/ml Extract A alone. Conclusion:Extract A contains caffeic acid with hemostatic effect, and its average value is 112.3 μg/g. Extract A can stimulate uterine smooth muscle, possibly by affecting M-receptors or N-receptors and non-selective β-receptors.