葛根芩连汤含药血清体外抗轮状病毒的实验研究

Studies on quality standard for Tibetan medicine Corydalis Herba

目的研究葛根芩连汤含药血清体外抑制轮状病毒(RV)的作用及其机制。方法采用观察CPE及MTT法,从抑制病毒生物合成、直接灭活病毒、抗病毒吸附三方面,测定葛根芩连汤含药血清对RV感染MA104细胞病变的作用。结果抑制病毒生物合成、直接灭活病毒方面,与相应浓度空白血清比较,各浓度葛根芩连汤含药血清与西药组含药血清使RV感染MA104细胞OD值明显升高(P<0.01);在抗病毒吸附方面,各浓度葛根芩连汤含药血清作用的细胞OD值明显高于同浓度空白组和西药组(P<0.01)。结论葛根芩连汤主要通过直接灭活轮状病毒以及抑制轮状病毒生物合成而发挥其抗病毒作用。

Objective To study the methods of quality evaluation and establish the quality standards of Tibetan medicine Corydalis Herba. Methods According to the methods recorded by the Appendix of Chinese Pharmacopoeia,the transverse section of root and powder were authenticated by microscopic method,and the protopine,used as control,in Corydalis Herba were identified and assayed by thin-layer chromatography（ TLC） and high performance liquid chromatography（ HPLC）. The TLC method was developed by using mixture of cyclohexane-dichloromethane-methanol（ 7 ∶ 2 ∶ 1） as adeveloping solvent on silica gel precoated plate. In HPLC method,protopine was separated on a reverse phase HPLC chromatographic column with a mixture of acetonitrile and triethylamine in acetic acid solution（ 24： 76） as the mobile phase with flow rate of 1. 0 ml/min,column temperature was maintained at 25℃ and the wave length was detected at 289 nm. Results The results showed that microscopic features of Corydalis Herba were distinctive,and the spots of protopine in TLC were clear with good resolution,which could be used to identify Corydalis Herba. The results of HPLC showed that protopine displayed good linearity within the range of 0. 91 ~ 464. 00 μg/ml. The average recoveries of protopine were 100. 6% with RSD of 0. 54%. The RSD value of intra-day and inter-day precision were less than 3%,while the contents of protopine was 0. 0974% ~ 0. 145%（ Average 0. 121%）. Conclusion The established method was sensitive,accurate with good repeatability,and it could be used to control the quality of Corydalis Herba.