| Cancer is one of the important reasons leading to death now. To find good therapeutic, less toxic and less side effect antitumor drugs from the natural products has become a major task in the field of pharmaceutical research today using modern science and technology. P. ramosissimus Poir is a folk medicine and hasn’t been collected in Chinese Pharmacopoeia. Now the related pharmaceutical studies have mainly focused on the separation and identification of the chemical constituents of its root and fruit. And there is no research reports related to the antitumor activity in addition to the folk medicine experience. In this paper, we investigated the antitumor and immunomodulatory effect of the extracts from the leaves of P. ramosissimus by the in vivo and in vitro experiments. Meanwhile, we determined the optimal harvest time of the leaves.At first, cold soak method was used to extract the fresh leaves of P. ramosissimus, and we got the ethanol and water extract of it. Then, the ethanol extract was isolated by systematic solvents. Combining with MTT methods, the inhibitory effect of these extracts, including water extract, ethanol extract, petroleum ether extract(MJZ I), ethyl acetate extract(MJZ Ⅱ) and n-butyl alcohol extract(MJZⅢ) to HepG-2, MGC-803 and Hela in vitro were assayed. The MJZⅡ showed significant anti-proliferation effect on tumor cells with dose-effect relation. IC50 values were 23.68、24.91、75.32μg·ml-1 respectively. The ethanol extract and MJZ I also exhibited certain inhibited activity. No obvious inhibited activities were observed in water extract and MJZIII. Expanding the scope of the cell lines, we inspected the proliferation inhibition of ethanol extract and MJZ Ⅱ in vitro, and found that they both have strong growth inhibition even to normal cells, et al. So we judged that they may have certain cytotoxicity. The mice bearing S180 were treated with MJZ Ⅱ and the inhibitory rate of tumor and immune organ index in vivo were evaluated. The results showed that MJZⅡ inhibit the growth of solid tumors in S180 mice obviously. The inhibitory rate of M. group was 49.38%. However, the sample had certain inhibitory effect on thymus (P<0.01). Using the MTT, neutral red and Griess method, we investigated the immunoregulatory activity of MJZⅡ in vitro. The results showed that MJZⅡ has certain inhibitory effect on mouse spleen lymphocyte proliferation with obvious dose-response relationship. But it has two-way adjustment effect on spleen proliferation induced by LPS and ConA. It didn’t have obvious effect on RAW264.7 cells engulfing neutral red dye and secreting NO. At last, we investigated the cell proliferation inhibition of the leaves collected from 5-10 month respectively aiming to determine the best harvest time combining with and antitumor activity as indicators. The results showed that the fresh leaves collected in July and August have the largest production, strongest activity and best quality. So July and August are the optimum harvest time of the leaves from P. ramosissimus.In conclusion, the extract from the leaves of P.ramosissimus (MJZ Ⅱ) both has significant antitumor activity in vivo and in vitro. It also has the inhibitory effect on the immune organ in vivo and can inhibit the spleen lymphocyte proliferation in vitro. But it has two-way regulating effect on spleen proliferation induced by LPS and ConA. P.ramosissimus has the great research value and development prospects as an antitumor natural medicine. But it still needs further research in the aspect of immunity activity. |
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