四种动物胆粉中胆酸类成分分析及药理作用比较

Composition determination of the cholic acid in bile powders from four different animals and comparison of their pharmacological actions

目的:采用HPLC法分别测定熊胆粉与猪胆粉、牛胆粉和鸡胆粉的胆酸类成分并进行药理作用的比较。方法:采用HPLC法对胆酸类成分分析。应用Lichrospher C18(4.6mm×250 mm,5μm)色谱柱,以乙腈-0.001 mol/L KH2PO4溶液为流动相,梯度洗脱(0～20 min,乙腈30%→30%;20～40 min,乙腈30%→40%;40～70 min,乙腈40%→50%),流速:1.0 ml/min,柱温30℃,DAD检测器,检测波长为215nm。以小白鼠为实验研究对象,利用扭体法,观察四种胆粉的镇痛作用,利用制备二甲苯致炎模型,观察四种胆粉的抗炎作用。结果:①胆酸类成分含量牛胆粉>鸡胆粉>熊胆粉>猪胆粉,四种胆粉中有相同的胆酸类成分,其中熊胆粉与牛胆粉的化学成分更为接近,共有峰重叠率达52.941%,相似度达0.540;②四种胆粉均能有效抑制二甲苯引起的小鼠耳朵肿胀,对炎症的抑制率依次为牛胆粉>熊胆粉>猪胆粉>鸡胆粉;③四种胆粉也能明显减少冰醋酸所致的小鼠扭体次数,延长潜伏期,作用强弱依次为熊胆粉>牛胆粉>鸡胆粉>猪胆粉。结论:熊胆粉与猪胆粉、牛胆粉和鸡胆粉相比较有相同的胆酸类成分,抗炎及镇痛作用接近,该方法为熊胆代用品的寻找提供了依据。

Objective：To determine the cholic acid fraction in the bile powders from bear,swine,cattle and chicken with HPLC technique,and compare the pharmacological actions among these components.Methods：HPLC technique was used to determine the cholic acid fraction by running on Lichrospher C18（4.6 mm×250 mm,5 μm） column with acetonitrile-water（0.001 mol/L potassium dihydrogen phosphate） as mobile phase in gradient elution for 0 to 20 min（acetonitrile 30%→30%）,20 to 40 min（acetonitrile 30%→40%）,and 40 to 70 min（acetonitrile 40%→50%）,respectively,at a flow rate of 1.0 ml/min,and at temperature of column by 30 ℃.DAD detector was set at 215 nm to detect the wavelength and reference wavelength.Acetic acid-induced mouse model writhing test was performed to observe the analgesic effects of the four kinds of bile powders,and another group of animal models were developed by induction of ear edema with dimethylbenzene to examine the anti-inflammatory potency of the four ingredients.Results：Although the diverse bile powders contained similar cholic acid,yet,the concentration ranked from bile powder of cattle to chicken′s,bear′s to swine′s.Of the 4 different bile powders,the chemical component in the bear bile powder appeared almost equal to that of ox,which led to an ratio of the common peak overlapping by 52.941% and semblance of 0.540.While the 4 sorts of bile powders were capable of producing anti-inflammatory effects on mice induced by dimethylbenzene,the inhibitory ratio of phlogosis was in sequence by ox bile powder〉bear bile powder〉porcine gall powder〉chicken bile powder.In addition,the 4 ingredients evidently reduced the frequencies of writhes of mice induced by acetic acid and led to prolonged latency,whereas the potency looked different,and the bear bile powder appeared the best,followed by components from cattle,chicken and swine.Conclusion：Among the 4 kinds of bile powders,the cholic acid fraction in the powder of bear looks comparative to that of swine′s and cattle′s to chicken′s,with similar analgesic and anti-inflammatory effects,and this findings will supply a basis for an alternative use of bear bile powder in the future.