| Oridonin was chosed as the research material, and Murine hepatoma cells(Hepa1c1c7) was used to study oridonin’s induced QR mechanism through the cell signal translation pathway. And Youhuaxiang, Zongsuoxie two oolong tea aroma and its main monomer were chosed as the research material, using Hepa1c1c7 and RAW264.7 cell models creened and separated active components. Evaluation of interactions between monomers and oridonin with monomers were also studied using Hep G2, MDA-MB-231 and HCT116 cell models. The results were as follows: 1.Oridonin’s induced QR mechanism(1) Oridonin presented the QR inducing potency with a CD value of 0.94 μg/m L. Treating 10μM PI3 K inhibitor significantly inhibit the activity of QR inducing, while treating 60μM ERK inhibitor significantly promote the activity of QR inducing, respectively 0.77 and 1.46 times as much as in the control. Oridonin don’t though p38, PKC and JNK pathway affecting QR activity.(2) During 0~12h, oridonin significantly stimulated the accumulation of Nrf2 to nuclear after treated 9 hours, which was 4.43 times as much as in the control. Besides, 5μg/m L oridonin significantly promoted Keap1 m RNA expression while reduced Keap1 protein in the cell, respectively 0.77 and 1.46 times as much as in the control.(3) 2.5μg/m L oridonin significantly promoted NQO1 m RNA and protein expression in Hepa1c1c7, respectively 3.47 and 2.56 times as much as in the control. 2. Volatile components of Youhuaxiang oolong tea anti-cancer/anti-inflammatory feature(1) Youhuaxiang oolong tea aroma when concentration of 15μg/m L inducing QR activity significantly. Aroma in 62.5~1000μg/m L concentration range of HCT116, MDA-MB-231 cells proliferation inhibition rate up to 68% and 70%. Aroma of 25μg/m L respectively with 5μM p38 and 2.5μM PI3 K inhibitors most inhibit QR activity the obvious after processing, the measured QR activity of aroma of individual treatment group V respectively 0.69 and 0.67 times.(2) Separation by chemical method, TLC method of 6 kinds of aroma products were 12.5~25μg/m L concentration within the scope of the induced QR multiply. Determination of linalool, nerolidol, indole, benzothiazole, limonene, benzyl alcohol, 2-amylfuran, diacetone alcohol 8 kinds of monomer, linalool most anti-cancer activity, within the scope of 44.4 to 55.5 μg/m L NO inhibition rate of 50%; according to the linalool: nerolidol: indole = 7.2:4.8:8.0 mixing ratio, mixed components within the scope of 12.5~25.0 μg/m L induced QR activity multiplication, namely within 25~ 50 μg/m L NO inhibition rate of 50%, with strong anti-cancer/anti-inflammatory activity. 3. Zongzuoxie oolong tea aroma anti-cancer/anti-inflammatory feature research(1) GC-MS analysis showed that the original gas and neutral component of alcohols in the majority, relative contents were 44.28% and 48.26% respectively; basic component contains more o-xylene, alcohols, relative contents were 28.48% and 17.88% respectively; acidic fraction of carboxylic acid compounds, o-xylene content was higher, the relative content of 22.55% and 14.25% respectively.(2) Induced QR multiplication concentration range: the original gas and neutral group is divided into early 12.5~25μg/m L, alkaline and acidic groups are divided into 25~50μg/m L, TLC separation product F1, F3 is 6.25~12.5μg/m L, secondary TLC separation product formula F1-1 of 6.25~12.5μg/m L, F1-2、F3-2 is 12.5~25μg/m L. NO inhibition rate of 50% concentration range: the original gas and neutral and alkaline groups are divided into 25~50μg/m L, acidic groups are divided into 12.5~25μg/m L, TLC separation product F1, F3 is 50~100μg/m L, secondary TLC separation product formula F1-2 was 25~50μg/m L,F1-1 and F3-2 were 50~100μg/m L. HCT116 inhibition rate of 50% concentration range: original, neutral group was divided into 125~250μg/m L. On MDA-MB-231 inhibition rate of 50% concentration range: neutral group was between 125~250μg/m L. 4. Anti-cancer interaction research between monomer and oridonin(1) Monomers in tea aroma:Indole, α-farnesene and methyl linolenate significantly induced QR at concentration of 42.42, 25.48, 30.52μg/m L, while the concentration of 50% NO inhibition were 85.42,59.7,74.43 μg/m L.(2) After impact Hep G2, HCT116 and MDA-MB-231 cells 24,48,72 h, Indole, α-farnesene and methyl linolenate all showed the highest proliferation inhibition rate of cancer cells in 72 h at 150 μg/m L. Highest Hep G2 cell proliferation inhibition rate were 33.01%, 79.15%, 82.35% respectively. Highest HCT116 cell proliferation inhibition rate were 71.90%, 99.30%, 99.77% respectively. Highest MDA-MB-231 cell proliferation inhibition rate were 71.90%, 99.30%, 99.77% respectively. Indole’s highest inhibition of proliferation effect was similar to methyl linolenate’s, and indole’s effect was weakest.(3) The strongest combination of synergistic inhibition effect of Hep G2 cell proliferation between monomers and oridonin with monomers were indole 150μg/m L+α-farnesene 150μg/m L, indole 100μg/m L+ oridonin 10μg/m L, the proliferation inhibition rate were 96.3%, 99.8%, CI value were 0.556, 0.391. The strongest combination of synergistic inhibition effect of HCT116 cell proliferation between monomers and oridonin with monomers were indole 144.54μg/m L + α-farnesene 116.38μg/m L, indole 144.54μg/m L+ oridonin 14.46μg/m L, the proliferation inhibition rate were 99.1%, 99.7%, CI value were 0.168, 0.067. The strongest combination of synergistic inhibition effect of MDA-MB-231 cell proliferation between monomers and oridonin with monomers were indole 144.54μg/m L + α-farnesene 116.38μg/m L, indole 144.54μg/m L+ oridonin 14.46μg/m L, the proliferation inhibition rate were 99.6%, 99.9%, CI value were 0.443, 0.500.(4) When CI value<1 at ED50, the largest DRI value of synergistic inhibition effect of Hep G2 cell proliferation between monomers and oridonin with monomers were indole+α-farnesene, indole+ oridonin, the DRI value were 3.08, 1.74 and 6.59, 2.41 respectively. the largest DRI value of synergistic inhibition effect of HCT116 cell proliferation between monomers and oridonin with monomers were α-farnesene+methyl linolenate, indole+ oridonin, the DRI value were 1.81, 1.81 and 2.34, 2.35 respectively. When CI value>1 at ED50, The strongest combination of antagonism inhibition effect of MDA-MB-231 cell proliferation between monomers and oridonin with monomers were α-farnesene+methyl linolenate, methyl linolenate+oridonin, CI value were 3.312, 1.958. The weakest combination were indole+α-farnesene, indole+oridonin, CI value were 1.550, 1.266. |
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