| Cedrus deodara is the general names of Pinaceae plants, its pine needles is one of 13 species medicinal pine needles, the chemical composition of pine needles of Cedrus deodara are mainly lignans、flavonoids、Organic acid, polysaccharide and Needles rubber, It has anti-tumor, antioxidant, antiviral and mosquito poison effect and many kinds of efficacy. In order to make full development and utilization of pine needles of Cedrus deodara,in this paper we carry on systematic research on total lignans of pine needles of Cedrus deodara.1.The study on establishing method of content determination for total lignans from pine needles of Cedrus deodara and optimizing the ectraction technology.Phosphomolybdic acid chromogenic method is used to determine the total lignans content,which uses holokiol as reference substance and detect the absorption value at 740 nm.On the basis of the central composite experimental design principles and one-factor test,the best extraction condition was selected by the method of response surface analysis with three factors and three levels, and the three influence factors were alcohol chroma, extraction time and solid- liquid ratio.the optimum extraction conditions of the total lignans from pine needles of Cedrus deodara as follows: еthanol concentration of 77.96%、ratio of liquid-to-solid 25:1、extraction time was at 1.78 h. Under these conditions, the yield of total lignans from pine needles of Cedrus deodara is up to 99.2 mg/g, which is close to the predicted value of 100.49 mg/g.2.The study of purification method of total lignans from Pine Needles of Cedrus deodara by macroporous resin. Use Pine Needles of Cedrus deodara total lignans adsorption rate and desorption rate as index,according to adsorption and desorption property of macroporous resins to lignans, S-8、D101、AB-8、X-5、NKA-9 and HPD-100 resins were compared for purifying total lignans from pine needles of Cedrus deodara. The results showed that AB-8 was the most appropriate resin. the optimal conditions were as follows: the sample amount was 6 bv(bed volume) at 1.5 ml/min flow rate,the sample concentration was 2.18 mg/ml. After saturation adsorption, remove water soluble impurities and residual liquid which is not adsorbed on the column by adding distilled water 5 BV at 1.5ml/min flow rate. Finally, desorb with 80% ethanol 4BV at 1 m L/min flow rate.Collect elution and boil away the liquid. The content of total lignans in the final production could reach 57.77%.3.The evaluation of the anti-tumor activity of total lignans and osthole extracted from Pine Needles of Cedrus deodara.The inhibitory effects of total lignans and osthole from Pine Needles of Cedrus deodara on A549、He La、Hep G2、HT-29 and MKN-45 cells were assessed by cck8 assay.,and screening out the most sensitive cancer cell on its total lignans and osthole. The results showed that total lignans and osthole has significant inhibition of cell proliferation on five tumor cells,the inhibitory effects of total lignans were A549>HeLa>HepG2>HT-29>MKV-45,the inhibitory effects of osthole were A549>HepG2>HT-29>MKV-45>HeLa, especially total lignans and osthole have significant inhibition of cell proliferation on A549 cell, IC50 of total lignans and osthole are 39.82μ g/mL and 35.68μ g/mL,respectively.The apoptosis and cell cycle was analyzed by Flow cytometry instrument,which probe the mechanism of total lignans and osthole induce apoptosis in human lung cancer A549 cells. It turned out that total lignans and osthole can also induced A549 cell apoptosis and total lignans causd the cell cycle of A549 accumulated in G0/G1 phase,osthole causd the cell cycle of A549 accumulated in G2 / M phase. |
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