骆驼蓬多糖提取及对秀丽隐杆线虫多聚谷氨酰胺聚集毒性的影响

目的探讨骆驼蓬多糖提取方法及对秀丽隐杆线虫株AM141肌肉细胞中多聚谷氨酰胺(Poly Q)聚集的影响。方法采用水提分级醇沉法对骆驼蓬多糖进行提取,并用苯酚-硫酸法对其进行含量测定。将骆驼蓬多糖加入培养基中,取样统计成虫肌肉细胞中Poly Q-YFP聚集所形成荧光点数目,以此评价骆驼蓬多糖对秀丽隐杆线虫株AM141中Poly Q聚集的影响。对活性较好的60%乙醇沉淀组分用大孔树脂进行纯化和活性再评价。结果 30%、60%、80%乙醇沉淀骆驼蓬多糖质量分数分别为20.07%、21.80%、19.98%,对秀丽隐杆线虫株AM141 Poly Q-YFP聚集所形成荧光点的抑制率分别为7.9%、17.00%和-1.72%。60%乙醇沉淀部分用大孔树脂纯化后,质量分数较高的10%、20%乙醇洗脱液中多糖质量分数分别为49.76%、38.26%,抑制率分别为35.25%和18.03%。结论利用秀丽隐杆线虫株AM141模型来评价骆驼蓬多糖对Poly Q聚集毒性的影响,方法简便,稳定可靠。且发现60%乙醇沉淀骆驼蓬多糖对秀丽隐杆线虫株AM141中Poly Q聚集的抑制作用最强,并有一定的量效关系。

Extraction of polysaccharides of Luotuopeng (Peganum harmala L.,Herb of Common Peganum) and its effect on toxicity of PolyQ accumulation in Caenorhabditis elegans

Objective To probe into the influence of polysaccharides of Luotuopeng (Peganum harmala L.,Herb of Common Peganum) on toxicity of PolyQ accumulation in the muscle cells of AM141 strain of Caenorhabditis elegans (C.elegans).Methods The polysaccharides of Luotuopeng were extracted by using water extraction followed by sequential alcohol deposition method,and the content of each precipitation was determined by using phenol-sulfuric acid method.The polysaccharides were added to medium,and then samples were collected for counting the number of fluorescence dots due to PolyQ-YFP accumulation in the muscle cells of C.elegans.The influence of the polysaccharides on toxicity of PolyQ accumulation in AM141 strain of C.elegans was reviewed.The precipitation precipitated by 60％ ethanol with higher activity was purified by using macroporous resin for re-reviewing the activity.Results The mass fractions of the polysaccharides precipitated by 30％ ethanol,60％ ethanol or 80％ ethanol were,respectively,20.07％,21.80％,19.98％.The inhibition rates of the polysaccharides on the fluorescence dots due to PolyQ-YFP accumulation in the muscle cells of C.elegans were,respectively,7.9％,17％ and-1.72％.After the precipitation precipitated by 60％ ethanol purified with macroporous resin,the mass fractions of the polysaccharides,in 10％ and 20％ ethanol elution with higher mass fractions,were,respectively,49.76％ and 38.26％,and the inhibition rates were,respectively,35.25％ and 18.03％.Conclusion The method for reviewing the influence of the polysaccharides on toxicity of PolyQ accumulation through model of AM141 strain of C.elegans is easy,stable and reliable.The polysaccharide precipitated by 60％ ethanol has the highest inhibitory effect on PolyQ accumulation with a dose-effect relationship.