目的 研究棉子糖抗衰老活性并探讨其作用机制。方法 棉子糖(150、200、250μmol·L^-1)加到大肠杆菌中孵育秀丽隐杆线虫后,显微镜下观察线虫身体弯曲频率、头尾摆动频率、体长,观察热应激刺激下线虫的存活时间;通过脂褐素荧光强度检测评价棉子糖对线虫抗衰老能力的影响;采用DAF-16核定位测定和超氧化物歧化酶-3(SOD-3)表达测定研究其抗衰老的相关机制。结果 与对照组比较,棉子糖3个浓度均使线虫的身体弯曲频率加快(P＜0.001);200μmol·L^-1棉子糖使线虫头尾摆动频率加快(P＜0.01);体长测试结果显示棉子糖3个浓度均对线虫生长发育无毒害作用。热应激测试中,与对照组比较,棉子糖3个浓度均显著延长线虫在热应激条件下的存活时间(P＜0.05)。脂褐素测试中,与对照组比较,棉子糖3个浓度均使线虫体内脂褐素含量显著降低(P＜0.01)。DAF-16核定位测定中,棉子糖3个浓度均促进DAF-16蛋白入核表达。SOD-3表达测定中,与对照组比较,棉子糖250μmol·L^-1浓度组的SOD-3蛋白表达水平显著上升(P＜0.05),与DAF-16核定位测定一并说明棉子糖抗衰老活性依赖ⅡS分子信号通路。结论 棉子糖能够促进线虫的活动能力,对其生长发育无毒害性,并能延缓线虫的衰老速度,促进DAF-16蛋白入核表达,提高SOD-3蛋白表达水平,其抗衰老机制依赖ⅡS分子信号通路。

Objective To study the anti-aging activity of raffinose and to explore its anti-aging mechanism.Methods Raffinose(150,200, and 250 μmol·L^-1) was added to Escherichia coli to incubate Caenorhabditis elegans, the body bending frequency, head and tail swinging frequency, and body length of the nematodes were observed under a microscope, and the survival time of the Caenorhabditis elegans under heat stress stimulation was observed. The effect of raffinose on the anti-aging ability of nematode was evaluated by lipofuscin, and the anti-aging mechanism was determined by DAF-16 nuclear localization and SOD-3 expression.Results In the body bending test, the three concentration groups of raffinose accelerated the body bending frequency of Caenorhabditis elegans campared with control group(P＜0.001). In the head-tail swing test, the raffinose of 200 μmol·L^-1 concentration group accelerated the head-tail swing frequency of nematodes campared with control group(P＜0.01). In the body length test, the three concentration groups of raffinose had no toxic effect on the growth and development of nematode. In the heat stress test, the three concentration groups of raffinose prolonged the survival time of Caenorhabditis elegans under heat stress(P＜0.05). In the lipofuscin test, the three concentration groups of raffinose decreased the content of lipofuscin in Caenorhabditis elegans. In DAF-16 nuclear localization assay, raffinose at three concentrations promoted the expression of DAF-16 protein into the nucleus. In the determination of SOD-3 expression, the level of SOD-3 protein expression increased in raffinose 250 μmol·L^-1 group,which together with DAF-16 nuclear localization assay showed that raffinose's anti-aging activity depended on ⅡS molecular signal pathway.Conclusion Raffinose can promote the activity of Caenorhabditis elegans, has no toxicity to its growth and development,and can delay the senescence of Caenorhabditis elegans, promote the expression of DAF-16 protein into nucleus and increase the expression level of SOD-3 protein. The anti-aging mechanism of raffinose depends on ⅡS molecular signal pathway.

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