酵母菌发酵对熟地黄生物活性的影响研究

发布时间：2018-06-26 18:20

本文选题：熟地黄 + 酵母菌发酵　；参考：《山东大学》2017年硕士论文

【摘要】：生地黄经炮制变成熟地黄,由一种清热凉血药变成一种补益药。前期研究表明熟地黄炮制过程中发生了美拉德反应,产生大量5-羟甲基糠醛(5-HMF),同时还原糖含量大幅度增加;经酵母菌发酵后其中5-HMF和小分子糖类被降解,对大鼠Ⅰ型糖尿病并发症的预防效果得到改善。这一结果提示5-HMF和小分子糖类可能不是熟地黄的有效成分,但是文献报道5-HMF和地黄寡糖具有多种生物活性。究竟5-HMF和地黄寡糖对熟地黄功效是否有贡献,需要进一步研究。本研究在前期研究的基础上,进一步评价酵母菌发酵对熟地黄生物活性的影响。主要内容如下:(1)熟地黄、发酵熟地黄和5-HMF对甲氨蝶呤化疗所致大鼠急性肝肾损伤的影响。结果发现熟地黄和发酵熟地黄均显著降低血清谷草转氨酶、血尿素氮、尿糖、尿N-乙酰-β-D-葡萄糖苷酶(NAG)的含量;而5-HMF仅显著降低尿NAG含量,对其它指标影响不显著。(2)长时间饲喂含熟地黄、发酵熟地黄和5-HMF饲料对正常大鼠的影响。发现熟地黄使大鼠尿液钙、镁和钠离子排出增加,粪便颗粒变大,排便量及粪氮排出量增加,而发酵熟地黄和5-HMF影响不显著。(3)5-HMF与人血清白蛋白的结合规律。荧光光谱法揭示5-HMF与白蛋白孵育不同时间,计算所得的结合位点数相似,均稍大于1;而HPLC检测发现5-HMF与白蛋白随时间增加其结合量逐渐增加,剩余5-HMF与孵育时间之间的关系符合反S型曲线;此外,质谱检测表明5-HMF与白蛋白的结合属于非共价结合。本论文结果表明:酵母菌发酵不影响熟地黄肝肾保护活性,小分子糖类和5-HMF不是熟地黄该作用的主要活性成分。小分子糖类可能与熟地黄致泻副作用有关,而5-HMF可能影响白蛋白的生物功能。本研究结果支持熟地黄经酵母菌发酵后应用。
[Abstract]:Rehmannia glutinosa processed into cooked Rehmannia glutinosa, from a medicine to cool blood into a tonic medicine. Previous studies showed that Maillard reaction occurred during the processing of Rehmannia glutinosa, resulting in a large number of 5-hydroxymethylfurfural (5-HMF), while the content of reducing sugar increased significantly, and 5-HMF and small molecular carbohydrates were degraded after fermentation by yeast. The prevention effect of diabetic complications in rats was improved. These results suggest that 5-HMF and small molecular carbohydrates may not be active components of Rehmannia glutinosa, but it is reported that 5-HMF and Rehmannia glutinosa oligosaccharides have many biological activities. The contribution of 5-HMF and Rehmannia glutinosa oligosaccharides to cooked Rehmannia glutinosa needs further study. On the basis of previous studies, the effect of yeast fermentation on the biological activity of Rehmannia glutinosa was further evaluated. The main contents are as follows: (1) the effects of cooked Rehmannia glutinosa, fermented Rehmannia glutinosa and 5-HMF on acute liver and kidney injury induced by methotrexate in rats. The results showed that both cooked Rehmannia glutamate and fermented Rehmannia glutamate significantly decreased the contents of serum glutamic oxaloacetic transaminase, blood urea nitrogen, urine sugar, urine N-acetyl- 尾 -Dglucosidase (nag), while 5-HMF only significantly decreased urinary nag content. There was no significant effect on other indexes. (2) the effects of long time feeding with cooked Rehmannia glutinosa, fermented Rehmannia glutinosa and 5-HMF feed on normal rats. It was found that cooked Rehmannia glutinosa could increase excretion of calcium, magnesium and sodium ions in urine, increase fecal granules, excretion of fecal nitrogen and excretion of feces, but had no significant effect on fermentation of cooked Rehmannia glutinosa and 5-HMF. (3) the binding of 5-HMF to human serum albumin. Fluorescence spectrometry revealed that the binding sites of 5-HMF and albumin were similar, and the binding amount of 5-HMF and albumin increased with the increase of time by HPLC. The relationship between the remaining 5-HMF and incubation time was in accordance with the anti-S-shaped curve, and mass spectrometry showed that the binding of 5-HMF to albumin was noncovalent. The results showed that yeast fermentation did not affect the liver and kidney protection activity of cooked Rehmannia glutinosa, and small molecular carbohydrates and 5-HMF were not the main active components of this action of Rehmannia glutinosa. Small molecular carbohydrates may be related to the side effects of diarrhea induced by Rehmannia glutinosa, while 5-HMF may affect the biological function of albumin. The results of this study support the application of cooked Rehmannia glutinosa after fermentation by yeast.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

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