酵母菌發(fā)酵對熟地黃生物活性的影響研究
本文選題:熟地黃 + 酵母菌發(fā)酵 ; 參考:《山東大學》2017年碩士論文
【摘要】:生地黃經(jīng)炮制變成熟地黃,由一種清熱涼血藥變成一種補益藥。前期研究表明熟地黃炮制過程中發(fā)生了美拉德反應,產(chǎn)生大量5-羥甲基糠醛(5-HMF),同時還原糖含量大幅度增加;經(jīng)酵母菌發(fā)酵后其中5-HMF和小分子糖類被降解,對大鼠Ⅰ型糖尿病并發(fā)癥的預防效果得到改善。這一結(jié)果提示5-HMF和小分子糖類可能不是熟地黃的有效成分,但是文獻報道5-HMF和地黃寡糖具有多種生物活性。究竟5-HMF和地黃寡糖對熟地黃功效是否有貢獻,需要進一步研究。本研究在前期研究的基礎(chǔ)上,進一步評價酵母菌發(fā)酵對熟地黃生物活性的影響。主要內(nèi)容如下:(1)熟地黃、發(fā)酵熟地黃和5-HMF對甲氨蝶呤化療所致大鼠急性肝腎損傷的影響。結(jié)果發(fā)現(xiàn)熟地黃和發(fā)酵熟地黃均顯著降低血清谷草轉(zhuǎn)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僅顯著降低尿NAG含量,對其它指標影響不顯著。(2)長時間飼喂含熟地黃、發(fā)酵熟地黃和5-HMF飼料對正常大鼠的影響。發(fā)現(xiàn)熟地黃使大鼠尿液鈣、鎂和鈉離子排出增加,糞便顆粒變大,排便量及糞氮排出量增加,而發(fā)酵熟地黃和5-HMF影響不顯著。(3)5-HMF與人血清白蛋白的結(jié)合規(guī)律。熒光光譜法揭示5-HMF與白蛋白孵育不同時間,計算所得的結(jié)合位點數(shù)相似,均稍大于1;而HPLC檢測發(fā)現(xiàn)5-HMF與白蛋白隨時間增加其結(jié)合量逐漸增加,剩余5-HMF與孵育時間之間的關(guān)系符合反S型曲線;此外,質(zhì)譜檢測表明5-HMF與白蛋白的結(jié)合屬于非共價結(jié)合。本論文結(jié)果表明:酵母菌發(fā)酵不影響熟地黃肝腎保護活性,小分子糖類和5-HMF不是熟地黃該作用的主要活性成分。小分子糖類可能與熟地黃致瀉副作用有關(guān),而5-HMF可能影響白蛋白的生物功能。本研究結(jié)果支持熟地黃經(jīng)酵母菌發(fā)酵后應用。
[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.
【學位授予單位】:山東大學
【學位級別】:碩士
【學位授予年份】:2017
【分類號】:R285.5
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