枸杞子多糖提取物LBP1C通过激活TFEB延缓衰老  

LBP1C Extracted From Lycium barbarum Delays Aging by Activating TFEB

作　　者：伍东立 乔新华[2] 韩文生 谢婷 时畅 黄雨云飞 孙传鑫 丁侃 陈畅 WU Dong-Li;QIAO Xin-Hua;HAN Wen-Sheng;XIE Ting;SHI Chang;HUANG Yu-Yun-Fei;SUN Chuan-Xin;DING Kan;CHEN Chang(School of Basic Medical Sciences,Southwest Medical University,Luzhou 646000,China;National Laboratory of Biomacromolecules,CAS Center for Excellence in Biomacromolecules,Institute of Biophysics,Chinese Academy of Sciences,Beijing 100101,China;College of Life Sciences,University of Chinese Academy of Sciences,Beijing 100049,China;Zhongshan Institute for Drug Discovery,Zhongshan 528400,China)

机构地区：[1]西南医科大学基础医学院,泸州646000 [2]中国科学院生物物理研究所生物大分子国家重点实验室,北京100101 [3]中国科学院大学生命科学学院,北京100049 [4]中国科学院上海药物研究所受体研究重点实验室,药物研究国家重点实验室,上海201203

出　　处：《生物化学与生物物理进展》2023年第8期1926-1936,共11页Progress In Biochemistry and Biophysics

基　　金：中国科学院战略性先导科技专项(B类)(XDB39000000);科技部重点研发计划(2022YFA1303000)资助项目。

摘　　要：目的探究枸杞子提取物LBP1C延缓人成纤维细胞及线虫自然衰老的效应及机制,以期解读《本草纲目》记载的枸杞子轻身不老、易颜色、变白的科学内涵。方法以人成纤维细胞及线虫自然衰老为模型,用LBP1C处理人成纤维细胞(复制性衰老模式细胞),检测P16、P21表达水平及β-gal染色指征衰老水平;用qPCR的方法检测转录因子GATA4及衰老相关分泌表型(senescence associated secretory phenotype,SASP)指标的mRNA水平;通过免疫荧光检测溶酶体生物发生和自噬的关键转录因子EB(transcription factor EB,TFEB)入核水平,通过Western blot检测LC3II/I的水平以指征LBP1C对自噬水平的影响;在细胞及线虫体系检测LBP1C对脂褐质水平进行检测。通过线虫身体在相同时间内的摆动次数和咽部吞咽次数评估LBP1C对线虫运动能力的影响。结果LBP1C处理的人成纤维细胞组中衰老相关指标(P16、P21表达水平及β-gal染色)相比于对照组显著下降,表明LBP1C可延缓细胞衰老;LBP1C还降低了细胞及线虫的脂褐质积累,并提高线虫运动及吞咽能力,促进线虫的健康从而抑制衰老。机制上,LBP1C处理组中TFEB入核比例显著升高,激活了自噬。自噬水平的提高降低了SASP的转录因子GATA4水平,从而降低了SASP相关的IL-1β及诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)水平。同时,自噬水平的提高使脂褐质降解加速,降低了细胞及线虫中脂褐质的积累。结论LBP1C通过激活TFEB提高了自噬水平,从而降低了SASP及脂褐质的积累,具有延缓衰老的效应,并促进了健康衰老。本研究初步揭示了枸杞子在抗衰老及抑制脂褐质从而潜在美白作用的效应及机制,为其进一步抗衰老和美白产品的研发提供了科学依据和理论基础。Objective This study aims to explore the effects and mechanisms of LBP1C extracted from Lycium barbarum on delaying the natural aging of human fibroblasts and nematodes,and provide scientific evidence and explanation for the anti-aging and skin whitening effects recorded in the“Ben Cao Gang Mu(Compendium of Materia Medica)”.Methods Human fibroblasts and C.elegans were used as natural aging models and treated with LBP1C.The expression levels of P16,P21 and the number of SA-β-gal staining positive cell were used to indicate the cell senescence level.The levels of GATA4,indicators related to the senescence associated secretory phenotype(SASP),transcription factor EB(TFEB)targeting genes and autophagy marker protein LC3 were detected by qPCR or Western-blot.The level of nuclear transcription of TFEB,a key regulatory factor for lysosomal biogenesis and autophagy,was detected by immunofluorescence in fibroblasts.The effects of LBP1C on lipofuscin levels in fibroblasts and C.elegans were respectively detected by lipofuscin kit and fluorescence microscope.The motor ability of LBP1C to C.elegans was evaluated through body bends and pumping analysis.Results Human fibroblasts and C.elegans were used as natural aging models in this study.Aging related indicators(P16,P21 expression levels and the number of SA-β-gal staining positive cell)in the LBP1C treatment group decreased markedly compared with the control group,which indicated that LBP1C delayed cell senescence.The proportion of TFEB nuclear translocation significantly increased in the LBP1C treated groups and activated autophagy.Detection of TFEB downstream genes showed that mRNAs of lysosomal enzymes,lysosomal membrane proteins and functionally related vesicular ATPases were all significantly upregulated in the LBP1C treatment groups.The increase in autophagy level autophagy reduced the level of SASP transcription factor GATA4,thereby reducing the levels of SASP related IL-1β and iNOS.Besides,the increase in autophagy level accelerated the degradation of lipofus

关 键 词：枸杞子 衰老 转录因子EB 自噬 衰老相关分泌表型 脂褐质 

分 类 号：Q255[生物学—细胞生物学]