Study On Antioxidant Activity Of The Bee Pollen Of Brassica Napus L. And Antitumor Activity Of The Dicranostigma Leptopodum (Maxim.) Fedde

1.Study on antioxidant activity of the bee pollen of Brassica napus L.Bee pollen of Brassica napus L. has a broad bioactivities, including enhancing body immunity, delaying consenescence, maintaining the digestive system, and preventing prostate degeneration. It is attributed partly to the antioxidants contained therein. In this paper the antioxidant activity of different fractions of bee pollen of Brassica napus L. were evaluated by offline DPPH and ABTS assay, online HPLC-DPPH assay and PC12cells oxidatived model by MTT assay. And based on this results, HPLC technique was used to screen and identify the antioxidant compounds in the bee pollen of Brassica napus L.The main research contents are as follows:(1) Eight kinds of extract fractions (fractionA-H) were obtained from the bee pollen of Brassica napus L. using reflux and macroporous adsorption resin. And then antioxidant activity of them were evaluated by offline radical scavenging capacity assay and online HPLC-DPPH assay. The results showed that all the eight kinds of extract fractions possessed free radical scavenging activity, whereas the60%ethanol extract fraction (fraction F) was the most appropriate fraction. The IC50of scavenging DPPH and ABTS free radicals were0.018mg·mL-1,0.015mg-mL-1, respectively.(2) Antioxidant activity of eight kinds of extract fractions were evaluated using the H2O2-induced PC12cells oxidatived model by MTT assay. The results showed that the survival rate of cells pretreated with fraction B reached90.0%, significantly higher than that of the model group with53.6%(P<0.001), and higher than that of the VE pretreated cells with85.0%(P<0.05). Sequentially, main activity compounds in ethyl acetate extract fractions was speculated by HPLC assays and comparing with chromatogram of the reference standard, it was found that the content of the three main flavonoids in the fraction B, including Kaempferol-3,4’-di-O-β-D-glucoside (KMG),Kaempferol-3-O-β-D-(2-O-β-D-glucosyl)glucopyranoside (KMP) and Kaempferol-3-O-β-D-glucoside (KMC), was distinctly higher than that of other fractions. This results implied that they were one of the main antioxidant active compounds in the bee pollen of Brassica napus L.2. Study on antitumor activity of the Dicranostigma leptopodum (Maxim.) FeddeDicranostigma leptopodum (Maxim.) Fedde has the efficacy of antimicrobial detoxification, subside a swelling and analgesic. These activities were attributed to the alkaloids contained in the Dicranostigma leptopodum (Maxim.) Fedde. Isocorydione was one of the active alkaloid of it. In this paper cell growth inhibitions of isocorydione on ten kind of cells line were analyzed by MTT assay and the effects of Isocorydione on cell proliferation and apoptosis of human lung adenocarcinoma A549cells line were studied. The main research contents are as follows:(1) Ten kind of tumor cells was treated with different concentrations of isocorydione. And then the cell growth inhibitions of isocorydione on ten kind of tumor cells line were determined by MTT assay. The results indicated that the isocorydione could inhibit the proliferation of ten tumor cells in a dose dependant manner. Whereas the inhibitory effect on the human renal carcinoma cell line786-0cells was strongest, and the IC50was51.3p.g-mL-1.(2) Annexin V-FITC/PI double staining assay was applied to detect apoptosis on A549cells cycle. And PI single staining assay was performed to examine the influence of the isocorydione by flow cytometry after A549cells were exposed to isocorydione48hours. The results of Annexin V-FITC/PI assay showed the apoptosis rates of high dose group and low dose group were (25.3±0.3)%,(10.69±0.35)%, respectively, higher than it of the negative control group (0.1±0.02)%,(P<0.05). indicated that induce apoptosis is one of the mechanism of the isocorydione inhibit A549cells. The PI single staining assay results showed that the isocorydione treated cells be arrest in S stage. Implied that the mechanism of the isocorydione induced A549cells apoptosis may be by inhibition of protein and DNA biosynthesis.