摘要
桦褐孔菌(Inonotus obliquus(Fr.)Pilat),是珍稀的大型药用真菌,在俄罗斯已有一百多年的应用历史,能治疗多种疾病。目前美国、韩国及日本正加大对该菌的研究,而我国尚处于研究的初始阶段。自美国的Humfeld H等对蘑菇的深层发酵进行了研究,并提出用发酵法来培养蘑菇的菌丝体后,研究者们纷纷用这种方法进行多种菌类的发酵研究。而国内外对桦褐孔菌的深层发酵研究甚少。对于其化学成分分析及药理学方面的研究尚未见报道。
本文首先对桦褐孔菌发酵菌粉的化学成分进行了包括水分、灰分、脂肪、蛋白质、氨基酸、三萜、总酚、多糖、还原糖、矿物元素在内的分析。结果表明桦褐孔菌菌粉营养丰富,含有蛋白25.73%,氨基酸14.50%,粗多糖28.55%。不饱和脂肪酸是优势脂肪酸,以亚油酸,油酸为主。还含有丰富的多糖、三萜类化合物、酚类物质和多种矿物元素。桦褐孔菌菌粉中砷和铅的含量分别为0.50μg/g和0.30μg/g,符合国家标准。桦褐孔菌菌粉多糖的主要单糖组成及质量分数分别为:阿拉伯单糖0.53%,甘露糖0.48%,葡萄糖10.75%,半乳糖2.44%。多糖产品中含有酸性多糖,糖苷键主要是α型。桦褐孔菌菌粉多糖提取的理想工艺条件为:温度为83℃,提取时间为2.2 h,液料比为33.3∶1,其多糖的提取率达到24.58%。
通过观察对正常小鼠血糖、糖耐量的影响;四氧嘧啶造模糖尿病小鼠的生长情况、血糖、血清胰岛素和肝糖原水平、对胰岛β细胞修复情况的影响、血清血脂、肝脏细胞修复情况及肝脏抗氧化能力的影响等方面考察了桦褐孔菌菌粉及其提取物的初步降糖、降脂及抗氧化作用。结果表明:桦褐孔菌菌粉及其提取物组分II具有良好的降血糖、降血脂和抗氧化能力。而且桦褐孔菌菌粉及其组分II可能降糖机理包括修复糖尿病小鼠受损胰岛β细胞、促进胰岛素分泌、增加肝糖原含量和提高糖尿病小鼠抗氧化能力。
Inonotus obliquus (Fr.) Pilat has been known for over one hundred years as a medicinal fungus in folk medicine, which can treat many refractory diseases. However, this fungus is just investigated in China at present though being explored extensively in American, Korea and Japan. Since American Humfeld H and other workers studied submerged culture of mushroom and put forward to culture mycelia of mushroom by this method, fungi have been cultured in succession. But the researches about I. obliquus are poor in this aspect. Until now it has not been studied chemically, manipulately, pharmacologically and so on.
This investigation characterized the chemical composition of dry matter of culture broth (DMCB) of I. obliquus in submerged culture, including moisture, ash, fat, crude protein, amino acid, triterpenoids, total phenol, polysaccharrides, reducing sugar and minerals. It’s shown that the DMCB of I. obliquus in submerged culture was rich of crude protein, amino acid and polysaccharides, the contents of which were 25.73%, 14.50% and 28.55%, respectively. And unsaturated fatty acids were prior to saturated fatty acids, in which oleic acid and linoleic acid were the main components. There were also many other nutrition components in it. Moreover, the content of As and Pb were 0.50 and 0.30μg/g, respectively, which met national standards. The monosaccharide components of polysaccharides extracted under the optimum condition were arabinose 0.53%, mannose 0.48%, glucose 10.75% and galactose 2.44%. Furthermore, there were acidic polysaccharides in the products and the glycosidic bonds wereα-types mainly. Simultaneously, an extraction temperature of 83℃, an extraction time of 2.2 h and a liquid-solid ratio of 33.3 were found to be optimal for polysaccharides extraction from DMCB of I. obliquus. By means of additional experiments, the adequacy of this model was conformed.
The antihyperglycemic and antilipidperoxidative effects of the DMCB of I. obliquus and its extractions were investigated in normal, glucose-induced hyperglycemic and alloxan-induced diabetic mice and the possible mechanism was also discussed. Treatment with the DMCB of I. obliquus and its extractions exhibited a mild hypoglycemic effect in normal mice, and failed to reduce the peak glucose levels after glucose administration. However, euglycemia was achieved in the DMCB of I. obliquus and its extractions treated mice after 120 min of glucose loading. In alloxan-induced diabetic mice, the DMCB of I. obliquus and its extractions showed a significant decrease in blood glucose level. Furthermore, the DMCB of I. obliquus and its extractions treatment significantly decreased serum contents of FFA, TC, TG and LDL-C, whereas effectively increased HDL-C, insulin level and hepatic glycogen contents in liver on diabetic mice. Besides, the DMCB of I. obliquus and its extractions treatment significantly increased CAT, SOD and GPx activity except for decreasing MDA level in diabetic mice. Histological morphology examination showed that the DMCB of I. obliquus restored the damage of pancreas tissues in mice with diabetes mellitus. The results revealed that the DMCB of I. obliquus and fraction II of DMCB possessed of significantly antihyperglycemic, antilipidperoxidative and antioxidant effects in alloxan induced diabetic mice.
引文
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