滑菇多糖的制备及其生物学效应
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摘要
本实验首次探讨了滑菇多糖(PNP)的制备工艺路线,及其对荷骨髓瘤BALB/c小鼠的抗肿瘤作用。实验结果表明:
     1.本实验条件下酶解结合水浸提联合工艺提取滑菇子实体多糖的最佳工艺路线为:浸提比1:10,纤维素酶(0.05%加酶量,pH 5.0,45℃)酶解60min,胰蛋白酶(0.05%加酶量,pH8.0,25℃)酶解45min,而后80℃保温搅拌提取90min。结果表明,酶解后热水浸提阶段并不是提取时间越长越好,一定时间后反而会出现负增长现象;联合工艺对滑菇多糖的提取率较单纯的酶解或水浸提,提取效率分别提高了37%和66%,效果显著,且易纯化,是行之有效的提取多糖的方法。
     2.联合工艺提取的滑菇多糖为纤维状灰褐色中性多糖,由D-甘露糖、D-葡萄糖和D-半乳糖三种单糖组成:经Sevag法联合胰蛋白酶去蛋白,超速离心分级得到的滑菇多糖,成分均一,多糖含量可达91.1%。
     3.动物实验表明,滑菇多糖能显著提高荷瘤小鼠的存活率;能提高荷瘤或正常小鼠的机体免疫功能。主要表现为:能有效地促进荷瘤小鼠和正常小鼠脾脏的生长和分化,增加脾脏的重量:能提高正常小鼠血清中TNF-α的含量,而且能有效降低荷瘤小鼠血清中的含量水平;能显著抑制荷瘤小鼠血清中IL-6的含量,但对正常小鼠没影响;能通过降低肿瘤组织中LDH的活性而对肿瘤的生长起抑制作用。
The best process route of extraction on Pholiota nemako polysaccharide (PNP) and its anti-tumor activity on tumorigenic BALB/c mouse were firstly studied. Three results showed as follow:
    1. An united method, that was extraction in water after enzymolysis (cellulase and trypsin), was used in this study. And the best extraction route was showed following: the proportion of digestion 1:10, cellulase enzymolysis (0.05% dosage, pH 5.0, 45 癈) sixty minutes, trypsin enzymolysis (0.05% dosage, pH 8.0, 25 癈 ) fourty-five minutes, then extraction keeping warm and stirring ninty minutes. The study result showed that the extractive efficiency was not as good as prolonging extractive time in the stage of hot water-extraction after enzymolysis. That was, the efficiency will decrease after some time. The extractive efficiency of this united method was better than the pure enzymolysis and pure extraction in hot water with promotion of 37% and 66% respectively. The efficiency was notable and polysaccharide extracted by this method was easy to be purified. All these showed that the extractive route was practicable and easy.
    2. PNP extracted by this united method was dust-coloured and neutral polysaccharide and were composed of D-mannose, D-glucose and D-galactose. PNP was a homogeneous ingredient after deproteinization by sevag-method and graded by ultracentrifugation. Its sugar content was up to 9 1 . 1 %.
    3. The animal experiment results showed that: PNP could promote the livability of tumorigenic mouse; could accelerate the growth and differentiation and put on weight of spleen; could increase the content of TNF- a in normal mouse' serum and decrease the content of tumorigenic mouse by different pathways probably; and could decrease the content of IL-6 in tumorigenic mouse' serum, but had no effection on normal mouse. In addition, PNP could inhibite the growth of tumor by decrease the activity of LDH in tumor-organization.
    In all, PNP could enhance the imrnunity-systerm of tumorigentic or normal mouses.
引文
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