茯苓多糖在秀丽隐杆线虫中的抗氧化和抗衰老作用及其作用机制  

Antioxidant and anti-aging effects and mechanism of Poria cocos polysaccharides in Caenorhabditis elegans

作　　者：宋祯彦 黄亚琦 王也彤 邓嘉 甄达贵 谭年花 成绍武 SONG Zhenyan;HUANG Yaqi;WANG yetong;DENG Jia;ZHEN Dagui;TAN Nianhua;CHENG Shaowu(Key Laboratory of Hunan Province for Integrated Traditional Chinese and Western Medicine on Prevention and Treatment of Cardio-Cerebral Diseases,Hunan University of Chinese Medicine,Changsha 410208,China;Jingzhou Miao and Dong Autonomous County Quality Measurement Inspection and Testing Center,Huaihua 418400,China)

机构地区：[1]湖南中医药大学中西医结合学院中西医结合心脑疾病防治湖南省重点实验室,湖南长沙410208 [2]靖州苗族侗族自治县质量计量检验检测中心,湖南怀化418400

出　　处：《中草药》2024年第4期1133-1144,共12页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81774129);国家自然科学基金资助项目(82004184);湖南省科技创新计划项目(2023RC3166);湖南省自然科学基金优秀青年项目(2023JJ20033);湖南省教育厅科研基金项目(23A0297);2023年度湖南省大学生创新创业训练计划项目(S202110541094)。

摘　　要：目的分析茯苓多糖成分对秀丽隐杆线虫体内抗氧化和抗衰老作用的影响,并阐明其潜在机制。方法将L4期线虫随机分为低、中、高剂量(10、20、40μg/mL)的茯苓多糖组和空白组进行培养;采用百草枯抗氧化实验、寿命实验、脂褐素测定实验、应激实验(紫外/高温)、生理功能实验(吞咽频率/摆动次数/产卵量)探究茯苓多糖对野生型秀丽隐杆线虫抗氧化和抗衰老作用;将突变体线虫分为茯苓多糖组(40μg/mL)和空白组,采用应激实验、寿命实验、线虫BZIP结构域蛋白1(protein skinhead-1,skn-1)绿色荧光蛋白的细胞定位检测、qRT-PCR实验、活性氧(reactive oxygen species,ROS)检测探究茯苓多糖的抗衰老的作用机制。结果茯苓多糖显著延长线虫的寿命,减少脂褐素的产生,增强对紫外线和热胁迫的抵抗力,提高咽部泵血频率和运动能力,显示出显著的抗衰老生物活性。茯苓多糖的抗衰老作用是通过skn-1信号通路介导的,而不依赖于胰岛素样受体β亚基(insulin-like receptor subunit beta,daf-2)和神经元乙酰胆碱受体亚基eat-2(neuronal acetylcholine receptor subunit eat-2,eat-2)信号通路。茯苓多糖增加了线虫中skn-1转录因子的核定位,上调了下游抗氧化基因谷胱甘肽S-转移酶4(glutathione S-transferase 4,gst-4)和gst-7的转录水平,并显著降低了细胞内ROS水平。然而,在skn-1突变体蠕虫中未观察到这些效应。结论茯苓多糖在秀丽隐杆线虫中表现出抗氧化和抗衰老的作用,其机制涉及调控skn-1转录因子及其下游抗氧化基因,最终降低ROS水平,增强机体的抗氧化应激能力。Objective To analyze the impact of Poria cocos polysaccharides(PCP)on antioxidant and anti-aging effects in Caenorhabditis elegans and elucidate its potential mechanisms.Methods L4 stage C.elegans were randomly divided to low-,medium-,high-dose(10,20,40μg/mL)PCP groups,and control group for cultivation.Anti-oxidant experiments,lifespan assays,lipofuscin determination,stress tests(UV/heat),and physiological function experiments(swallowing frequency/number of swings/egglaying amount)were conducted to investigate the anti-oxidant and anti-aging effects of PCP on wild-type C.elegans.Mutant C.elegans were divided into PCP group(40μg/mL)and control group.Stress tests,lifespan assays,cell localization detection of C.elegans BZIP domain protein 1(skn-1)green fluorescent protein,qRT-PCR experiments,and reactive oxygen species(ROS)detection were performed to explore the mechanisms of PCP in anti-aging.Results PCP significantly extended the lifespan of C.elegans,reduced lipofuscin production,enhanced resistance to UV and heat stress,increased pharyngeal pumping frequency,and improved locomotor activity,demonstrating significant anti-aging biological activity.The anti-aging effect of PCP was mediated through the skn-1 signaling pathway,independent of the insulin-like receptor subunit beta(daf-2)and neuronal acetylcholine receptor subunit eat-2 signaling pathways.PCP increased the nuclear localization of skn-1 transcription factor in C.elegans,upregulated the transcription levels of downstream anti-oxidant genes,including glutathione S-transferase 4(gst-4)and putative glutathione S-transferase 7(gst-7),and significantly reduced intracellular ROS level.However,these effects were not observed in skn-1 mutant worms.Conclusion PCP exhibit antioxidant and anti-aging effects in C.elegans,involving the regulation of the skn-1 transcription factor and its downstream anti-oxidant genes,ultimately reducing ROS level and enhancing the organism’s anti-oxidant stress capacity.

关 键 词：茯苓多糖 秀丽隐杆线虫 抗衰老 抗氧化 BZIP结构域蛋白1 

分 类 号：R285.5[医药卫生—中药学]