| The germination a wonderful phenomenon of life, is the first developed stage of life and the most active biological stage of life. After the seeds have sucked adequate water, a series of physiological and metabolic activities occur, such as the recovery and the biochemical metabolism of cells, which changes the nutritional compositions significantly. Appropriately germinating tartary buckwheat sprout, which has a high nutritional value is a newly developed sprout vegetable. This paper used germinated tartary buckwheat powder as materials and studied on several aspects of changes in nutritive substances and the main physiological activity during germination.1. The effect of tartary buckwheat’s germination processes on flavonids content and CHI activity changes.The content of total flavonoids in powder of tartary buckwheat sprouts germinated from 0 to 6 days would be determined in this paper by using aluminum color visible spectrophotometry methodology. With the increase of germination days, the content of total flavonoids increased, and the highest content was on the sixth day(26.6±1.0mg/g). It was found that the main components of tartary buckwheat flavonoids were rutin and quercein through documents previously published, so HPLC method was built to detect the content of rutin and quercein in tartary buckwheat sprouts(1~6d). And with the extension of germination(1~6d), the content of rutin raised while the content of quercein reduced. One thing to be mentioned was that the proportion of rutin and quercetin was close to 1:1 on the third day. During the germination, the activity of CHI took the same situation as the change of the content of flavonoids. The study revealed that an increase in CHI activity would effectively promote the content of flavonoids.2. Study on the extraction and determination of tartary buckwheat trypsin inhibitor andγ-aminobutyric acid(GABA)Tartary buckwheat trypsin inhibitor(TBTI) was extracted by (NH4)2SO4 segmentation precipitation, and the crude enzyme’s activities was analyzed by choosing 50% inhibition reaction system, so the optimum concentration of trypsin was 0.25mg/mL, the optimum substrate(BAPNA) concentration was 0.06mM, the best doses of inhibitor was 10μl, the time of heat preservation was 10min, the inhibitor activity unit was 0.3675U, the specific inhibitor activity was 3.8994 U/mg.The content of GABA in powder of tartary buckwheat sprouts germinated from 0 to 6 days and bran would be measured by ninhydrin colorimetric method. As the days of germination increased, the content of GABA got highest on the third day, and then decreased. The content of GABA in buckwheat sprouts was higher than bran, and these datas were in some rule.3. Study on antitumor activities of flavonids in tartary buckwheat sprouts(1) With MTT colorimetric method, the different anticancerous effects of flavonoids ethanol extract from tartary buckwheat sprouts and the rutin(Ru) and quercetin(Qu) mixed according to the data detected by HPLC and themselves on the model containing human breast cancer MCF-7 cell line was analyzed in this assay. It was showed that the total flavonoids ethanol extract from tartary buckwheat sprouts powder had the inhibition effect on MCF-7 cell, and the extraction from the 3rd had the highest inhibition rate, which was 38.58±4.98%, and it was much higher than that of control group(P<0.01). The main flavonoid compositions in ethanol extract from tartary buckwheat sprouts were quercetin and rutin, and their mixture(QR) had better inhibition rate than total flavonids, and the ethanol extraction of mixture for 3days(the proportion of rutin and quercetin was close to 1:1), which inhibition rate was 62.22±2.74%, they had a good co-effect. The law of ethanol extract from tartary buckwheat sprouts had a similar inhibition effect with the analogue pattern of standard quercetin and rutin. MCF-7 cells were treated with different concentration gradient of monomer Qu and Ru, and it was calculated that Qu IC50 was 8.55μg/mL, Ru IC50 was 785.26 ug/mL. It was showed in the result that Qu could obviously inhibit the proliferation of MCF-7 cells at the lower concentration and in a dose-dependent manner. To determine whether the QR had better effect of inhibition than monomer, the same quality of Qu and Ru with K′3 was selected. The results indicated that inhibitory action was QR>Qu>Ru, and the inhibitory effect of QR group was higher than that of positive control(5-FU), and it proved that K′3 had similar function as 5-FU. Foremore, Qu and Ru can reinforce inhibitory effects on tumor growth.(2) To observe whether tartary buckwheat sprouts (TBS) had antiangiogenesis effect, eggs incubated for 7 days was hatched and the TBS ethanol of different days and mixture were added to the carrier which was on the surface of CAM respectively. Two days later, the states of angiogenesis on the CAM were observed. It showed that both the crude and pure TBS could inhibit neovessels’growth, and the inhibition effect increased with the increase of the concentration of flavonids. The inhibition effect of crude group was not obvious compared with mixture, and also Qu had better effect than Ru, and this had a conformable trend with tumor cell proliferation.4.A preliminary study on antitumor mechanism of flavonids in tartary buckwheat sproutsCell cycle and apoptotic rate were investigated by flow cytometry, and flavonids of TBS powder ethanol extract K1, K3, K5 and mixture of Ru and Qu ethanol solution K′1, K′3, K′5 was selected to disposal MCF-7 cells within 48h, and they were compared with the control group. The result showed that in flavonids group, the cancer cells were blocked in S and G2/M phases, but the cancer cells in Go/Gl phase decreased. Meantime, apoptotic rate increased with the increase of the concentration of flavonids, and it indicated that flavonids could induce apoptosis of MCF-7 cells. K′1, K′3, K′5 was compared with the control group, and it showed that K′3 had a noticeable inhibition effect (P <0.01), which blocked the most cancer cells in S phases, and had the highest apoptotic rate.5. Study on the effect of the activities of tartary buckwheat sprouts on immune response in miceTo determine the effect of TBS(K5~K6) which had the highest content of flavoniods, and mixture(QR) on the immune function in experimental mice. Through the determination of delayed allergy(DTH), serum anibody level, NK cells active function test(LDH method) and the spleen lymph cells transformation of ConA induced mouse(MTT method), it was found that, compared with control group, both TBS and flavonids monomer could improve the immunitive function of DTH, hemoglobin amount, the NK cell’s activity and thelymphocyte trasformation rate significantly in all those treatment group. From the result of first three tests, it was found, Ru had better effect than Qu. The results showed that the anti-cancer effect of the total flavones was due to the improvement of the body immunity, and the anti-cancer effect of two single flavones were interpromoting. |
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