巨噬细胞膜包被的纳米粒对小鼠肝脏缺血/再灌注损伤的影响和机制研究  

Mechanisms of macrophage membrane coated nanoparticles in alleviating hepatic ischemia/reperfusion injury in mice

作　　者：陈政 易竹君[2] 傅奎 CHEN Zheng;YI Zhu-jun;FU Kui(Department of Hepatobiliary Surgery of Traditional Chinese Medicine Hospital Dianjiang of Chongqing,Dianjiang 400830,China;Department of Hepatobiliary Surgery of the Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China)

机构地区：[1]重庆市垫江县中医院肝胆外科,重庆408300 [2]重庆医科大学附属第二医院肝胆外科,重庆400010

出　　处：《中国现代普通外科进展》2021年第8期589-593,共5页Chinese Journal of Current Advances in General Surgery

基　　金：四川省教育厅重点项目(17ZA0132)。

摘　　要：目的:探讨巨噬细胞膜包被的纳米粒(M-NPs)对小鼠肝缺血/再灌注损伤(HI/RI)的影响及机制。方法:提取C57BL/6小鼠腹腔巨噬细胞(PMs)、制备PLGA微球(NPs),合成M-NPs并进行鉴定。构建小鼠HI/RI模型,将小鼠分为对照组(Control)、小鼠HI/RI组(HI/RI)以及小鼠HI/RI+M-NPs组(M-NPs)。Control组不做处理,M-NPs组小鼠在建模后立即通过尾静脉注射300 mg/kg的M-NPs;HI/RI组在建模后注射等量生理盐水。24 h后,采集小鼠肝组织、外周血和Kupffer细胞。HE染色检测肝组织病理改变,检测血清ALT、AST和TBil水平,ELISA检测血清LPS水平,Western blotting检测Kupffer细胞TLR4、M yD88、p-NF-κB以及NF-κB的蛋白表达。结果:成功制备M-NPs。HI/RI组小鼠肝组织病理损伤明显,大量炎症细胞浸润;M-NPs组肝组织虽仍有一定程度的肝损伤,但与HI/RI组相比,肝组织结构相对正常,肝细胞病理改变较轻,浸润的炎症细胞也明显减少。血清ALT、AST、TBil、TNF-α、IL-6和LPS水平的变化趋势与HE染色结果一致。与HI/RI组相比,M-NPs组小鼠Kupffer细胞中TLR4、MyD88和p-NF-κB的蛋白表达显著降低。结论:M-NPs可能通过中和小鼠HI/RI时产生的LPS,抑制LPS与Kupffer细胞膜TLR4的结合,抑制Kupffer细胞TLR4/NF-κB通路的活化,抑制炎症反应,进而减轻小鼠HI/RI。Objective:To investigate the effect and mechanism of macrophage membrane coated nanoparticles(M-NPs)on hepatic ischemia/reperfusion injury(HI/RI)in mice.Methods:Extracted the peritoneal macrophages(PMs)from C57 BL/6 mice and prepared PLGA microspheres(NPs),and synthesized M-NPs.A mouse HI/RI model was established and the mice were divided into Control group,HI/RI group and HI/RI+M-NPs group(M-NPs).Control group did not treat,and M-NPs group mice received 300 mg/kg of M-NPS via tail vein immediately after modeling.The HI/RI group was injected with the same amount of normal saline after modeling.After 24 h,the mouse liver tissue,peripheral blood and Kupffer cells were removed:HE staining detected the pathological changes of liver tissue,Detected the levels of serum ALT,AST and TBil.Western blotting detected the protein expressions of TLR4,MyD88,p-NF-κB and NF-κB in Kupffer cells.Results:In HI/RI group,the pathological damage of liver tissue was obvious and a large number of inflammatory cells infiltrated,Although liver tissue in M-NPs group still has some degree of liver injury,compared with HI/RI group,liver tissue structure is relatively normal,liver cell pathological changes are mild,and infiltrated inflammatory cells are also significantly reduced.The changes of serum ALT,AST,TBil,TNF-α,IL-6 and LPS levels were consistent with HE staining results.Compared with HI/RI group,the protein expressions of TLR4,MyD88 and p-NF-κB in Kupffer cells of mice in M-NPs group were significantly decreased,suggesting that M-NPs inhibited the activation of inflammatory pathway in Kupffer cells.Conclusion:M-NPs may inhibit the combination of LPS and TLR4 in Kupffer cell membrane by neutralizing LPS produced during HI/RI in mice,and inhibit inflammatory reaction by inhibiting activation of TLR4/NF-κB pathway in Kupffer cell,thus reducing HI/RI in mice.

关 键 词：肝 缺血再灌注损伤 仿生纳米粒 小鼠 

分 类 号：R657.3[医药卫生—外科学]