Reseach On Antitumor Activity Of Dryocrassin ABBA Treatment From Dryopteris Crassirhizoma

Dryopteris crassirhizoma belonging to the Dryopteridaceae. D.crassirhizoma has rhizome medicine, which functions indetoxification, deworming and hemostasis. It is also widely used in clinical treatments of common cold and heat rash. D.crassirhizoma is known to be rich in natural ingredients, containing phloroglucinol derivatives,flavonoid and polysaccharide etc. The content of phloroglucinols are high. Total phloroglucinols extract from D.crassirhizoma has significant anticancer activity to multiple transplanted animal tumor. Dryocrassine ABBA is phloroglucinol derivative extracted from D.crassirhizoma. Few researches were carried out on Dryocrassine ABBA’s purification process and cellular antitumor.In this study, wild D.crassirhizoma were used as materials to study the extraction and purification process of phloroglucinols from D.crassirhizoma and explore the potential anticancer activity of Dryocrassin ABBA towards human Hep G2 cells for the first time.(1) Dried D.crassirhizoma was powdered by disintegrator and then sieved through a mesh(40 mesh). 200 g of the powder was added into 2000 m L of ethanol(Volume fraction was 80%). Then ultrasonic(800 W) was applied to assist extraction for 30 min under 30 and it was repeated for ℃3 times. With n-hexane extraction, take the upper liquid. The filtered solutions were gathered and concentrated to dryness by removing the ethanol solvent using a rotary evaporator device at 45℃.(2) The separation conditions of total phloroglucinols from D.crassirhizoma was researched by DM-130 macroporous resin in this study. Using adsorption rate, elution rate, purity of phloroglucinols as indexes to investigate adsorption and desorption of phloroglucinols with DM-130 resin. By studying the optimum loading concentration of total phloroglucinols and maximum sample volume were 1.5 mg/mL and 210 mL respectively. Then using the concentration of 80% ethanol eluting target components, the flow fate was 1.0mL/min. The DM-130 resin performed perfect adsorption and desorption efficiency. After purification, After dynamic separation on DM-130 resin column adsorption, the contents of total phloroglucinols in the product was 14.17-fold increased.(3) After purification we using HPLC analysis the results, the content of Dryocrassin ABBA in total phloroglucinols is highest. Dryocrassin ABBA has significant anticancer activity to Human tumor cells. Preliminary experiments showed Hep G2 cells were most sensitive to Dryocrassin ABBA.(4) In this study, the apoptotic effects of Dryocrassin ABBA on the Hep G2 cells was investigated. We using MTT assay measured Hep G2 cells viability. Dryocrassin ABBA induced Hep G2 cells apoptosis. The results of Western blot showed that Dryocrassin ABBA up-regulated Bax and inhibited Bcl-2 expression which led to caspase cascade reaction, finally induced Hep G2 cell apoptosis. Overall, these findings provide evidences that Dryocrassin ABBA may induce apoptosis in Hep G2 cells through mitochondrial pathway.