| Honey is food and medicine, as well as a commonly used excipient in preparation of Chinese medicines. Before being used as excipient, it always needs to be processed into refined honey during which its stickiness enhanced. However, during processing Chinese medicines, it is necessary for refined honey to be diluted with water and then being mixed with medicine slices for stir-frying. So it is difficult to explain the processing mechanism of refined honey only by enhancing the stickiness.Previous studies showed that the hydroxymethylfurfural (HMF) contents in honey greatly increased after being processed. Furthermore, in consideration of the increased color of refined honey, we hypothesized that Maillard reaction and Caramelization occurred during the processing. The products may be the material basis to enhance or change the bioactivities of medicine slices. In order to test and verify this hypothesis, in this dissertation, the physicochemical properties and anti-oxidant activities of the products obtained from the fructose, glucose and HMF system with or without amino acids were evaluated. In addition, the refined honey (R-h), glutamic sodium-fortified refined honey (GR-h) (adding glutamic sodium to enhance Maillard reaction) and fermented solutions of GR-h (FGR-h) (adding yeast to reduce sugars and HMF) were prepared to compare and study the physicochemical properties and in vitro and in vivo bioactivities. The main results are as follows:1.The effects of heat processing on fructose, glucose and HMF systems:When fructose and glucose respectively underwent heat processing under the condition of 100℃and pH 4.0, the absorbance at 420nm (A420) of reaction system together with contents of HMF and DPPH radical scavenging activity increased gradually. After 72 hours, A420, HMF content and DPPH radical scavenging activity of the fructose system were increased by 9.09,17.38 and 0.81 times respectively compared with the glucose system. After reaction with 19 kinds of amino acids (at the mole concentration proportion of 50:1) for 12 hours, A420 of fructose and glucose systems were increased by 0.09-2.27 and 0.68-30.43 times respectively compared with the systems without amino acids. The majority of amino acids had little effect on HMF content and DPPH radical scavenging activity of the fructose system (except cysteine, proline and tryptophan) but had great effects on the glucose system (except cysteine) of which the HMF content and DPPH radical scavenging activity were increased by 0.32-16.13 and 0.45-1.31 times respectively. The A420 and DPPH radical scavenging activity of HMF system which was heated under the condition of 100℃ and pH 4.0 were increased by 10.90 and 6.24 times compared with the unheated. Except for tryptophan, all reactions with the other 18 kinds of amino acids further significantly enhanced the A420 and DPPH radical scavenging activity of HMF system.2. The effect of amino acid salt and yeast fermentation on the refined honey: Refining honey by means of adding glutamic sodium at 120℃ for 2 hours led to the darker color of honey sample based on the Lab color model. HMF content in fresh honey (H) was only 0.084mg/g, simple heating process increased HMF content in honey significantly, to be 1.124mg/g, and glutamic sodium further increased HMF level but only by 25%. For the R-h, the DPPH scavenging activity and total antioxidant capacity were increased by 6.32 and 2.97 times respectively compared with the H, and that of GR-h were increased by 2.30 and 2.66 times respectively compared with the R-h. The sugar content of H was 794.500mg/g, the highest among all the honey samples. Heating process with or without amino acid salt resulted in the decreased sugar contents in honey. As far as GR-h, yeast fermentation led to degradation of the most of sugars and HMF, reducing from 530.180 and 1.408mg/g to 98.160 and 0.335mg/g, respectively. However, the color and anti-oxidant activity had little change.3. The preventive effects of different refined honey samples on side effects of chemotherapy:Kunming mice were subcutaneously injected with methotrexate (MTX) (5mg/kg body weight) to induce intestinal mucositis. Using GSH (0.13g/kg body weight) as positive control, the preventive treatment groups were respectively given H, R-h, GR-h and FGR-h (2.6g/kg body weight). It was found that the treatment groups except R-h displayed improved aspects in body weight, food intake and white blood cell counts decrements, total activity in open field test, spleen index and the morphology of duodenum compared with MTX model group. Among all the treatment groups FGR-h showed the best improvement effect. As far as biochemical indicators, MTX model mice had higher activity of myeloperoxidase (MPO) and inducible nitric oxide synthase (iNOS) (p<0.001, p<0.001) and higher levels of malonaldehyde (MDA) and protein carbonyl (p< 0.001, p< 0.001) than normal control (NC) mice. All the treatment groups showed improved effects among which FGR-h had the most significant effect with all significant levels were p<0.001.4. The effect of different refined honey samples on the proliferation of human leukemia cell line K562:By the method of MTT test, the effects of MTX with different concentrations and the mixture of MTX and different honey samples on the viability of human leukemia cell line K562 were investigated. The results showed that the cell viability gradually declined with the concentration of MTX increased. When compared with the application of MTX alone, the combined application of MTX at 0.05μmol/L and GR-h or FGR-h at 0.5mg/mL exerted significant inhibitory effect on the growth of cell line K562 (p<0.01, p<0.01). However, H and R-h at the same concentration showed little effect. When at 0.1mg/mL all honey samples did not showed significant activity.In conclusion:Maillard reaction greatly contributes to the changes of correlated components and activities of refined honey. The application of Maillard reaction and yeast fermentation can improve the efficacy of refined honey, raising its application value in alleviating intestinal mucositis induced by chemothearphy. |
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