Studies On Toxicology Of HELA Which Treat With Ethanol Extracts Of Bombyx Batrytieatus

Bombyx Batryticatus as a traditional Chinese medicine, the application has been a long time. Modern pharmacological studies have shown that Bombyx Batrytieatus has Anti-inflammatory, antibacterial, anti-cancer, antithrombotic, anticonvulsant, and hypnotic effect. Studies on cytoxicity of Hela cell which treat with alcohol extract of Bombyx Batrytieatus in vitro ,could provide the basis for anticancer effect of Bombyx BatrytieatusIn this paper, though the morphological observation, MTT method, DNA Ladder, RT-PCR, Western blotting, we detected Cell viability and DNA fragmentation, expression changes of Bax and Bcl-2 of Hela cells which treat with different concentrations alcohol extraction.Results: After 24h, the morphology of Hela which treat with different concentration of drug (3mg/ml, 4mg/ml, 5mg/ml and 6mg/ml) did not change significantly; The same concentration, after 48h, the cells began to shrink, especially, when treat with the concentration of 4mg/ml, 5mg/ml and 6mg/ml, most cells have shrunk.In the test of cell viability, The viability of Hela which treat with different concentration of drug (3mg/ml, 4mg/ml, 5mg/ml and 6mg/ml, 7mg/ml, 8mg/ml, 9mg/ml, 10mg/ml, 11mg/ml) is promote by drug, after 24h; The same concentration, after 48h, the viability of Hela begins to diminish, when treat with the concentration of 6mg/ml, 7mg/ml 8mg/ml, 9mg/ml, 10mg/ml, 11mg/ml, drugs began to show inhibition of cell growth, inhibition rate is less than 50%, and the inhibition rate increased with the concentration; The same concentration, after 72h, the inhibition of cell growth is more obvious, the inhibition rate is more than 50%; When treat with the concentration of 9mg/ml, 10mg/ml, 11mg/ml, the inhibition rate is up to 70%In the test of DNA ladder, drug induce fragmentation of DNA in HeLa cells after treatment for 36h, HeLa cells are treated with different concentration of drug (3mg/ml, 4mg/ml, 5mg/ml, 6mg/ml and 7mg/ml); especially, when treat with the concentration of 7mg/ml, most cells DNA have fractured.RT-PCR and Western blotting analysis ofβ-actin, Bax, Bcl-2 RNA and protein expressions in HeLa cells after treatment for 36 h, HeLa cells are treated with different concentration of drug (3mg/ml, 4mg/ml, 5mg/ml, 6mg/ml); RT-PCR and Western blotting can detectβ-actin and Bax expression in RNA and protein of treatment group, the expression of Bcl-2 in RNA and protein can be detected only in control group. This shows that the drugs can inhibit Bcl-2 expression in RNA and protein.