聚乙二醇修饰的四氧化三锰纳米粒子对软骨细胞炎症模型的影响  

Effect of polyethylene glycol-modified manganese oxide nanoparticles on chondrocyte inflammation model

作　　者：张盛清 蔡金宏 黄兰丽 高明 赵劲民[1,2,4,5] Zhang Shengqing;Cai Jinhong;Huang lanli;Gao Ming;Zhao Jinmin(Guangxi Engineering Center in Biomedical Materials for Tissue and Organ Regeneration,Nanning 530021,China;Collaborative Innovation Centre of Regenerative Medicine and Medical Bioresource Development and Application,Nanning 530021,China;Pharmaceutical College,Guangxi Medical University,Nanning 530021,China;Guangxi Key Laboratory of Regenerative Medicine,Nanning 530021,China;Department of Orthopedics Trauma and Hand Surgery,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西组织器官修复医用生物材料工程技术研究中心 [2]广西医科大学再生医学与医用生物资源开发应用协同创新中心 [3]广西医科大学药学院 [4]广西再生医学重点实验室 [5]广西医科大学第一附属医院创伤骨科手外科,南宁530021

出　　处：《广西医科大学学报》2022年第2期181-187,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81972120);广西科技基地和人才专项(桂科AD19254003);广西医科大学青年科学基金资助项目(No.GXMUYSF201905);广西自然科学基金面上项目(No.20209XZSFAA159134)。

摘　　要：目的:探讨聚乙二醇修饰的四氧化三锰纳米粒子(Mn_(3)O_(4)-PEG NPs)对过氧化氢(H_(2)O_(2))诱导的软骨细胞炎症模型的抗炎抗氧化作用。方法:合成Mn_(3)O_(4)-PEG NPs并对其进行表征。通过CCK-8法和细胞活死染色法检测Mn3O4-PEG NPs对软骨细胞活性的影响。体外检测Mn_(3)O_(4)-PEG NPs的拟超氧化物歧化酶活性(SOD)、拟过氧化氢酶酶活性(CAT)和清除羟基自由基(·OH)的作用。将由H2O2诱导的软骨细胞炎症模型与Mn_(3)O_(4)-PEG NPs共培养后,通过逆转录获得cDNA,再用RT-qPCR法对共培养细胞中的白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)的表达水平进行测量。结果:经TEM、XRD、FTIR、ZETA电位和细胞毒性检测,结果证实Mn_(3)O_(4)-PEG NPs成功合成,在具有良好分散性的同时,也有着出色的生物相容性。体外活性氧(ROS)清除实验证明Mn_(3)O_(4)-PEG NPs具有三重拟酶活性。RT-qPCR结果显示,与对照组相比,Mn_(3)O_(4)-PEG NPs可以降低软骨细胞炎症模型中的TNF-α、IL-1β和IL-6表达。结论:Mn_(3)O_(4)-PEG NPs成功合成,在具有三重拟酶活性的同时,保持了较好的生物相容性,可以降低软骨细胞中因过量ROS所引起的炎症因子表达上升,是一种有着良好前景的防治骨关节炎(OA)的纳米酶。Objective:To investigate the antioxidant effect of polyethylene glycol-modified Mn3 O4 nanoparticles(Mn_(3)O_(4)-PEG NPs) on hydrogen peroxide(H_(2)O_(2)) induced chondrocyte inflammation.Methods:Mn3 O4-PEG NPs were synthesized and characterized.The toxic effects of Mn_(3)O_(4)-PEG NPs on chondrocytes were examined by CCK-8 assay and living-death staining.The superoxide-mimetic dismutase(SOD) activity,catalase(CAT)-mimetic enzyme activity,and hydroxyl radical(OH) scavenging effects of Mn3 O4-PEG NPs were examined in vitro.After co-culture of chondrocyte inflammation model induced by H_(2)O_(2) with Mn3 O4-PEG NPs,cDNA was obtained by reverse transcription,and the expression levels of interleukin-1β(IL-1β),interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) in the co-cultured cells were measured by RT-qPCR.Results:The results of TEM,XRD,FTIR,ZETA potential and cytotoxicity tests confirmed that Mn_(3)O_(4)-PEG NPs were successfully synthesized,with a good biocompatibility and dispersity.Reactive oxygen species(ROS) scavenging experiments in vitro showed that Mn_(3)O_(4)-PEG NPs had triple enzyme-mimetic activity.PCR results showed that compared with the control group,Mn_(3)O_(4)-PEG NPs could reduce IL-1β,IL-6 and TNF-α expressions in chondrocyte inflammation model,which had a significant effect.Conclusion:Mn_(3)O_(4)-PEG NPs nanoparticles are successfully synthesized,with a good biocompatibility and triple enzyme-mimetic activity,and can reduce the rise in the expression of inflammatory factors due to overdose ROS in chondrocytes,which is a promising nanozyme for the treatment of osteoarthritis(OA).

关 键 词：四氧化三锰 纳米酶 氧化应激 骨关节炎 

分 类 号：R318.08[医药卫生—生物医学工程]