| Eupatorium adenophorum (E. adenophorum) Spreng is a species of flowering plant in the daisy family known by many common names, including eupatory, sticky snakeroot, crofton weed, and Mexican devil. After the introduction as a ornamental plant to USA in 1960s, it has spread worldwide. It was first inadvertently introduced to Yunnan around 1940 from the China-Burma border. Now it distributes in the Province of Sichuan, Guizhou, Guangxi and Tibet. E. adenophorum has become one of the most serious harm invasive species in our country. E. adenophorum is a worldwide noxious weed and has caused tremendous detrimental effects on agriculture, forestry, rangeland and natural ecosystem since it invaded in southwesten China. At present, there were no effective control methods. The aim of this study is to extracted euptox A from E. adenophorum and to explore the bioactivity. The main contents and results were as follows:Euptox A was extracted from E. adenophorum by immersion as follows: ultrasonic-methanol extraction, Ethyl acetate extraction, Silica column Chromatography method, thin-layer Chromatography (TLC), D101 Macroporous Resin, Quantification and toxin purity were demonstrated by high performance liquid chromatography (HPLC). The purity of the toxin we had extracted was over 98%.The acaricidal activity of the euptox A, a cadenine sesquiterpene from E. adenophorum against S. scabiei and P. cuniculi was tested in vitro. A complementary log-log (CLL) model was used to analyze the data of the toxicity tests in vitro. The results showed euptox A had strong toxicity against mites, killing all S. scabiei at 3 and 4 mg/mL (m/v) concentration, while 4 mg/mL euptox A was also found to kill all P. cuniculi within a 4 h period. Similarly,2,3 and 4 mg/mL concentration of euptox A had strong toxicity against S. scabiei, with median lethal time (LT50) values at 0.687, 0.526,0.326 h, respectively.3 mg/mL and 4 mg/mL showed strong acaricidal action against P. cuniculi; the LT50 values were 0.693 h and 0.493 h, respectively. The median lethal concentration (LC50) values were 1.068 mg/mL for Scabies mite and 0.902 mg/mL for P. cuniculi in 2 h. The results indicate that euptox A has strong acaricidal activity and may exploit as novel drugs for the effective control of animal acariasis.The apoptosis inducing effect of the 9-Oxo-10,11-dehydroageraphorone (euptox A) on HeLa cells was examined by MTT assay. Meanwhile, the mechanism was analyzed by flow cytometry and QRT-PCR. The results suggest that euptox A had significant antitumor activity against the three tumor cell lines in vitro in a dose-dependent manner. When the concentration of euptox A was at 500g/mL, the percent inhibition of human lung cancer A549 cells, HeLa cells and Hep-2 cells were 76.42%,68.30% and 79.05% respectively. The fifty percent inhibitory concentration (IC50) of euptox A for the three tumor cell lines were 369,401 and 427μg/mL (A549, HeLa, and Hep-2 cells, respectively). Flow cytometry results suggest that euptox A could effectively inhibit the proliferation of HeLa cells, arrest the cell cycle transition from S to G2/M phase, can’t continue to complete the cell cycle activity (mainly form four times and mitosis), and make the cell proliferation. The QRT-PCR detection results showed that the expression levels of caspase-10 gene in HeLa cells were increased after incubation with euptox A for 24 h, and not showed dependence with euptox A dose. Firstly increased and then decreased with the increase of euptox A dose. In addition, the expression levels caspase-10 gene in treated with 30μg/ml of euptox A reach highest. However, in HeLa cells the caspase-3 gene was showed only minor signs decreased after incubation with euptox A for 24 h. |
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