In this study, we showed that crude extract of Anisomeles indica (AI-EtE) expressed its toxicity to HeLa cells with an IC50 dose of 38.8 µg/mL and to zebrafish embryos with malformations, lethality and hatching inhibition at 72-hpf at doses higher than 75 µg/mL. More interestingly, flow cytometry revealed that AI-EtE significantly promoted the number of cells entering apoptotic. Accordingly, the transcript levels of BAX, CASPASE-8, and CASPASE-3 in the cells treated with AI-EtE at IC50 dose were 1.55-, 1.62-, and 2.45-fold higher than those in the control cells, respectively. Moreover, treatment with AI-EtE caused cell cycle arrest at the G1 phase in a p53-independent manner. Particularly, percentages of AI-EtE-treated cells in G1, S, G2/M were, respectively 85%, 6.7% and 6.4%; while percentages of control cells in G1, S, G2/M were 64%, 15% and 19%, respectively. Consistent with cell cycle arrest, the expressions of CDKN1A and CDNK2A in AI-EtE-treated cells were up-regulated 1.9- and 1.64-fold, respectively. Significantly, treatment with AI-EtE also decreased anchorage-independent growth of HeLa cells. In conclusion, we suggest that Anisomeles indica can be considered as a medicinal plant with a possible use against cervical cancer cells; however, the used dose should be carefully monitored, especially when applying to pregnant women.

Many  species in the family Lycopodiacea possess highly potential medical substances, notably are Huperzine A of Huperzia serrata and alkaloids isolated from Lycopodium clavatum(α-Onocerin , Lycopodine,..). Various members of this family are found throughout of Vietnam and a few researches on its diversity as well as alkaloid isolation has been conducted, however, there is a lack of toxicology studies on this subject. In this study, three crude extracts (labeled AI.1, AI.2 and AI.3), which come from Lycopodium sp.collected from 3 different regions of Vietnam, have been tested on 2 toxicological models: primary neural cells derived from Swiss white mouse (in vitro) and Zebrafish (Danio rerio) embryos (in vivo). Our results show that the all three extracts are relatively low in toxicity, IC50 value on primary neural cell ranging from 800 - 1100 mg/L, while LC50 of fish embryos at 100 - 300 mg/L. AI.2 has the least effect of viability of both cells and embryos compare to the others. This data give the prove showed that the safety using Lycopodium as tradional medicine.