骨髓间充质干细胞衍生的细胞外囊泡改善重症急性胰腺炎小鼠肺组织损伤  

Extracellular vesicles derived from bone marrow mesenchymal stem cells improve lung tissue injury in mice with severe acute pancreatitis

作　　者：索朗玉珍 胡倩[3] 杨月 康鸿鑫 张蜀 万美华[3] Suolang Yuzhen;HU Qian;YANG Yue;KANG Hongxin;ZHANG Shu;WAN Meihua(Department of Emergency Medicine,West China Hospital,Sichuan University,Chengdu,Sichuan 610041,P.R.China;Department of Emergency,Tibet Autonomous Region People’s Hospital,Lhasa,Tibet 850000,P.R.China;West China Center of Excellence for Pancreatitis,Institute of Integrated Traditional Chinese and Western Medicine,West China Hospital,Sichuan University,Chengdu,Sichuan 610041,P.R.China)

机构地区：[1]四川大学华西医院急诊科,成都610041 [2]西藏自治区人民医院急诊科,拉萨850000 [3]四川大学华西医院中西医结合中心,胰腺炎中心,成都610041

出　　处：《华西医学》2024年第11期1718-1727,共10页West China Medical Journal

基　　金：中国博士后第74批面上项目(2023M742507);四川省科学技术厅自然科学基金项目(2023NSFSC1765);四川省医学会医学科研课题计划(S23065);四川大学华西医院专职博士后研发基金(2023HXBH006)。

摘　　要：目的 探讨骨髓间充质干细胞(bone marrow mesenchymal stem cell, BMSC)分泌的细胞外囊泡(extracellular vesicle, EV)对重症急性胰腺炎(severe acute pancreatitis, SAP)小鼠肺组织损伤的效应及潜在机制。方法 选择24只无特定病原体级雄性C57BL/6小鼠和小鼠原代肺微血管内皮细胞(pulmonary microvascular endothelial cell, PMVEC)。将小鼠分为假手术组、SAP组、BMSC组,每组8只。将小鼠原代PMVEC分为模型组[牛磺胆酸钠(sodium taurocholate, NaTC)组]、BMSC-EV组、对照组。提取并鉴定健康小鼠BMSC及BMSC-EV,建立SAP小鼠模型,将BMSC-EV尾静脉注射到SAP小鼠体内或体外干预小鼠原代PMVEC,观察胰腺和肺组织病理损伤,血清淀粉酶、脂肪酶、炎症因子[肿瘤坏死因子-α(tumor necrosis factor α, TNF-α)、白细胞介素-6(interleukin-6, IL-6)]等变化,肺组织和PMVEC炎症因子、内皮细胞屏障相关蛋白[钙黏蛋白E(E-cadherin)、紧密连接蛋白ZO-1、细胞间黏附分子(intercellular cell adhesion molecule-1, ICAM-1)]等表达,以及PMVEC间紧密连接情况,以探索BMSC-EV对SAP小鼠胰腺、肺组织和体外PMVEC的影响。结果 BMSC具有成骨、成软骨、成脂分化的潜能,其衍生的EV具有典型的杯状结构,直径在60~100 nm,BMSC-EV表达EV阳性蛋白TSG101和CD63,不表达阴性蛋白Calnexin。与假手术组比较,SAP组胰腺发生病理损伤(P<0.05),血清淀粉酶、脂肪酶、IL-6、TNF-α水平均上调(P<0.05);而BMSC-EV改善SAP小鼠胰腺组织损伤(P<0.05),下调外周血清淀粉酶、脂肪酶、IL-6、TNF-α水平(P<0.05),并上调IL-10水平(P<0.05)。除此之外,SAP小鼠还表现肺组织病理损伤(P<0.05),较高水平的炎症因子TGF-β、IL-6(P<0.05),较多的中性粒细胞和巨噬细胞浸润(P<0.05),表达较低水平的E-cadherin、ZO-1(P<0.05)和较高水平的ICAM-1(P<0.05);BMSC-EV改善了SAP小鼠肺组织病理损伤、炎细胞浸润、炎症因子水平和屏障蛋白表达(P<0.05),并抑制ICAM-1表达(P<0.05)。在Objective To investigate the effect and potential mechanism of bone marrow mesenchymal stem cells(BMSCs)-derived extracellular vesicles(EVs)on lung tissue injury in mice with severe acute pancreatitis(SAP).Methods A total of 24 specific pathogen free grade male C57BL/6 mice and primary mouse lung microvascular endothelial cells(PMVECs)were selected.The mice were divided into sham group,SAP group,and BMSC group,with 8 mice in each group.The mouse primary PMVECs were divided into model group[sodium taurocholate(NaTC)group],BMSC-EV group,and control group.Extraction and characterization of healthy mouse BMSCs and their derived extracellular vesicles(BMSC-EVs)were conducted.A mouse model of SAP was established,and BMSC-EVs were injected into SAP mice by tail vein or intervened in PMVECs in vitro,to observe the pathological damage of pancreatic and lung tissues,the changes of serum amylase,lipase,and inflammatory factors[tumor necrosis factorα(TNF-α),interleukin-6(IL-6)],the expression of inflammatory factors of lung tissues and PMVECs,and the endothelial cell barrier related proteins[E-cadherin,ZO-1,intercellular cell adhesion molecule-1(ICAM-1)],and tight junctions between PMVECs to explore the effects of BMSC-EVs on pancreatic and lung tissues in SAP mice and PMVECs in vitro.Results BMSCs had the potential for osteogenic,chondrogenic,and lipogenic differentiation,and the EVs derived from them had a typical cup-shaped structure with a diameter of 60-100 nm.BMSC-EVs expressed the extracellular vesicle-positive proteins TSG101 and CD63 and did not express the negative protein Calnexin.Compared with the mice in the sham group,the SAP mice underwent significant pathological damage to the pancreas(P<0.05),and their serum amylase,lipase,inflammatory factor IL-6,and TNF-αlevels were significantly up-regulated(P<0.05);whereas,BMSC-EVs markedly ameliorated the pancreatic tissue damage in the SAP mice(P<0.05),down-regulated the levels of peripheral serum amylase,lipase,IL-6 and TNF-α(P<0.05),and up-regulated the level of

关 键 词：间充质干细胞 重症急性胰腺炎 肺微血管内皮细胞 紧密连接 急性肺损伤 

分 类 号：R576[医药卫生—消化系统]