黄精衍生的纳米囊泡抑制脂多糖诱导的炎症因子产生及其分子机制  

Molecular Mechanisms Underlying Inhibition of LPS-induced Inflammatory Cytokine Production by Nanovesicles Derived from Rhizoma Polygonati

作　　者：曹腾辉 龙兴旺 刘林 王刚林 李伟[1,2] CAO Teng-Hui;LONG Xing-Wang;LIU Lin;WANG Gang-Lin;LI Wei(Wenzhou Medical University School of Laboratory Medicine and Life Sciences,Department of Biochemistry and Molecular Biology;Key Laboratory of Medical Genetics of Zhejiang Province,Wenzhou 325035,Zhejiang,China)

机构地区：[1]温州医科大学检验医学院(生命科学学院)生物化学与分子生物学系 [2]浙江省医学遗传学重点实验室,浙江温州325035

出　　处：《中国生物化学与分子生物学报》2025年第2期249-259,共11页Chinese Journal of Biochemistry and Molecular Biology

摘　　要：植物外泌体样纳米囊泡是指从植物中分离的含有脂质、蛋白质、RNA和各种小分子的球状脂质囊泡,具有抗炎、抗肿瘤、抗氧化和药物载体等方面功效,然而黄精衍生的纳米囊泡的功能尚未见报道。本文利用超速离心和密度梯度离心,首次从黄精中获得了外泌体样纳米囊泡(rhizoma polygonati exosome-like nanovesicles,RP-EVs),并对其表征和抗炎功能进行探究。结果:RP-EVs是主要带负电荷,平均粒径为166.5±3.3 nm的球状脂质囊泡。细胞摄取实验显示,RP-EVs可以被巨噬细胞吞噬。qPCR以及ELISA研究表明,RP-EVs可以抑制脂多糖(lipopolysaccharides,LPS)刺激引起的白细胞介素6(interleukin 6,IL-6)、白细胞介素1β(IL-1β)和肿瘤坏死因子α(tumor necrosis factor-alpha,TNF-α)(P<0.0001)的升高。不仅如此,活性氧(reactive oxygen species,ROS)及DPPH清除实验证实,RP-EVs具有一定的抗氧化功能(P<0.05)。进一步探究其机制,利用免疫荧光以及Western印迹检测发现,RP-EVs是通过IκBα/NF-κB信号通路抑制细胞核因子p65(nuclear factor kappa-B p65,NF-κB p65)的入核转运(P<0.01)及磷酸化(P<0.001),进而调控炎症因子表达。经动物实验,将RP-EVs腹腔注射至小鼠体内48 h,主要定位于小鼠的肝和脾。最后,利用腹腔注射LPS构建小鼠急性炎症模型,通过qPCR以及ELISA法检测炎症因子水平,发现RP-EVs可以缓解LPS引起的小鼠血清和脾中的炎症因子的表达(P<0.05)。总之,本论文首次分离获得了RP-EVs,并揭示了其抗炎功能和潜在的机制,将为中药来源的纳米囊泡的功能探究提供一定的参考,为炎症性相关疾病的治疗提供新的策略。Plant-derived exosome-like nanovesicles refer to spherical lipid bilayer vesicles isolated from plants that contain lipids,proteins,RNAs,and various small molecules.These nanovesicles exhibit diverse biological activities,including anti-inflammatory,anti-tumor,antioxidant,and drug delivery properties.However,the functional characteristics of nanovesicles derived from rhizoma polygonati remain unexplored.In this study,exosome-like nanovesicles derived from rhizoma polygonati(referred to as RP-EVs)were successfully isolated using ultracentrifugation and density gradient centrifugation.Their physicochemical properties and anti-inflammatory functions were systematically characterized.Our results show that RP-EVs are predominantly negatively charged,with an average particle size of 166.5±3.3 nm,and are spherical lipid vesicles.Cellular uptake assays demonstrated that RP-EVs can be phagocytized by macrophages.qPCR and ELISA experiments revealed that RP-EVs can inhibit the elevation of interleukin 6(IL-6),interleukin 1β(IL-1β),and tumor necrosis factor-alpha(TNF-α)induced by lipopolysaccharide(LPS)stimulation(P<0.0001).Additionally,reactive oxygen species(ROS)and 2,2-diphenyl-1-picrylhydrazyl(DPPH)scavenging assays confirmed that RP-EVs exhibit antioxidant properties(P<0.05).Further investigation of the underlying mechanisms through immunofluorescence and Western blotting revealed that RP-EVs inhibit the nuclear translocation(P<0.01)and phosphorylation(P<0.001)of nuclear factor kappa-B p65(NF-κB p65)via the IκBα/NF-κB signaling pathway,thereby regulating the expression of inflammatory mediators.In animal experiments,intraperitoneal injection of RP-EVs into mice for 48 hours showed predominant localization in the liver and spleen.Finally,an acute inflammatory mouse model was established via intraperitoneal injection of LPS.qPCR and ELISA analyses demonstrated that RP-EVs alleviated the expression of inflammatory factors in both the serum and spleen of LPS-treated mice(P<0.05).In conclusion,this study isolated RP-

关 键 词：黄精外泌体样纳米囊泡 炎症因子 活性氧 细胞核因子p65 

分 类 号：Q291[生物学—细胞生物学]