| Marine resource is an important natural medicine treasure for mankind. Owing to the unique living environment, marine organisms are endowed with a huge numbers of compounds which take strange and novel chemical structure as well as the high activitiy. By taking Pegasus laternarius Cuvier as the object to research, the separation and purification of the active compounds against cervical cancer cells was performed. Moreover, the apoptosis mechanism of cervical cancer cell line Hela was also probed, In addition, the preliminary pharmacodynamics experiments of the crude extract of Pegasus laternarius Cuvier were carried out to investigate the activities of anti-fatigue, immune regulation, anti-inflammatory, anti-impotence, anti-asthma and so on.The integral fish of Pegasus laternarius Cuvier was extacted with ethanol (95%, v/v) and water successively. Four kinds of components (A1, A2, A3 and A4) were obtained by the gradient elution of silica gel column from the ethanol extract, the growth inhibition against human cervical cancer cell line Hela was determined by the method of MTT, which showed that the components of A2 and A3 have stronger antitumor activity with dose and time dependent. Six kinds of components (B21, B22, B23, B31, B32 and B33) were obtained by the gradient elution from the components of A2 and A3 in a silica gel column. The further results showed that the component of B32 has the strongest anti-tumor activity to Hela cells. Three kinds of component (C321, C322 and C323) were obtained by the further separation step from the component of B32 in a Sephadex LH-20 gel column, showed the result that the component of C322 has the strongest anti-tumor activity to Hela cells.The apoptosis mechanism of the component of C322 against the cervical cancer cell line HeLa was also investigated. The result upon the detection method of MTT showed that the component of C322 has stronger anti-tumor activity against Hela cells, with IC50 value were 90.06±2.53 μg/mL、43.63±1.19 μg/mL at 24 h and 48 h, repectively. The results upon the method of AO/EB staining showed that the HeLa cells display the typical apoptosis characteristics of nucleus pycnosis and nuclear burst obviously after the treatment by the component of C322 for 48 h. The result shows also demonstrated that the component of C322 could induce the retardation in G0/G1 phase and (or) S phase through PI staining for detecting cell cycle distribution. Annexin V/PI experiments exhibited that the apoptosis of the HeLa cells were associated with the treating time and drug concentration of the component of C322 for 48 h. The result detection of the Caspase 3/8/9 enzyme activity showed that the enzyme activity were boosted by comparing with the control group, which indicated that the apoptosis induced by the component of C322 may be attributed to the activation of the downstream Caspase 3, which was resulted activation of the upstream Caspase8/9.The experiments of the turn bar test and weight loading swimming of mice showed that the water extract of Pegasus laternarius Cuvier has protential anti-fatigue effect. The experiments in vivo of the water and ethanol crude extacts showed no the exact immune enhance effect but in vitro. The ethanol extract, showing no significant effect on the ear swelling of mice which induced by xylene, indicated that it has no obvious anti-inflammatory effects appeared. Furthermore, the two crude extracts showing no significant effect on the weight of sex organs and accessory sex organ of castrated rats, indicated that they have no obvious anti-impotence effects. Moreover, the experiments in vitro showed that the ethanol extract had no significant effect on the airway smooth muscle contraction in guinea pigs induced by acetylcholine and histamine, indicated that it has no antagonism of airway smooth muscle contraction effects. |
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