| Hedyotis diffusa Willd (HDW), Rubiaceae Hedyotis diffusa dry grass, widely distributed in subtropical regions, are cultivated in China’s Fuji an, Guangdong, Jiangxi, Jiangsu, Anhui and other places. The character is slightly bitter taste and sweet-cold, and it has anti-tumor, anti-inflammatory, antibacterial and enhances immune effect certified by pharmacological studies. In recent years, with more and more attention in terms of anti-tumor effect, the effective parts and compounds of HDW’s anti-tumor have been found. It is reported that triterpenoids ursolic acid and oleanolic acid as two of the ingredients in this traditional Chinese medicine have significant anti-tumor effect. This paper selected the HDW from different areas of Fujian Province as the research object, and the fat-soluble ingredients were extracted by heating reflux method and using ethanol as extraction solvent. In order to provide a more scientific and reliable method for the quality control of HDW from Fujian province, the determination of ursolic acid and oleanolic acid were conducted on reversed-phase high performance liquid chromatographic. In addition, the work of screening of antitumor cell and anti-inflammatory effects, extraction parts by water decoction, ethanol extraction, and extraction separation by methanol from HDW, has been conductedThe fat-soluble components of HDW are extracted by ethanol heating reflux extraction. The content of ursolic acid and oleanolic acid of HDW as index components are0.038%-0.058%and0.201%-0.241%respectively, which measured by reversed-phase high-performance liquid chromatography. The results show that the ursolic acid and oleanolic acid content of the highest collected is from Zhangzhou, in seven palaces such as Fuzhou, Fuqing, Longyan, Zhangzhou, Sanming and Ningde, which provides a more scientific and reliable basis for quality control of HDW.The extract and chemical parts in inhibiting the proliferation of tumor cells in vitro is evaluted by morphological observation and MTT determination method. The results show that the ethanol extract, petroleum ether part for human gastric cancer cell line AGS, ethyl acetate and n-butanol parts for the human lung adenocarcinoma cell line A549,petroleum ether part all have a strong inhibition of proliferation in vitro, and showing a significant dose-dependent, namely inhibition rate with the drug concentration increasing. It also can be seen from the figure that water extract for SW579and A549, ethanol extract, water extract, petroleum ether, water parts for the A549and n-butanol parts for AGS, ethanol extract, ethyl acetate, n-butanol parts for DU145have some degree of inhibition, and showing a dose-dependent, but the effect is not obvious. The experiments of mice U14tumor model show that the ethanol extract of HDW can inhibit the growth of U14tumor, and showing a dose-dependent, improving the thymus index and spleen index, prompting the effect on inhibiting tumor growth in vivo.The results from xylene-induced ear edema model experiment show that ethanol extract can effectively reduce the xylene-induced ear edema within the concentration range of250-500mg·kg-1, and the water extract can play a significant pharmacodynamic effect for the mice ear swelling within the concentration range of800-2000mg·kg-1. In addition, it is found that four parts of ethanol extract have effect for the xylene-induced mice ear swelling, and the most significant effect is the n-butanol part.The results of experiments in rat granuloma model show that when concentration of the extractant of HDW is200,300mg.kg-1, the effect is obvious. |
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