白细胞介素10工程化修饰人脐带间充质干细胞优效治疗炎症性肠病  

Interleukin-10 engineered human umbilical cord mesenchymal stem cells for superior treatment of inflammatory bowel disease

作　　者：冯乙芮 高天芸 王亚萍 黄亚红[4] 王斌[2] Feng Yirui;Gao Tianyun;Wang Yaping;Huang Yahong;Wang Bin(Clinical Stem Cell Research Laboratory of Gulou Hospital Affiliated to Nanjing University School of Medicine,School of Life Sciences of Nanjing University,Nanjing 210008,Jiangsu Province,China;Clinical Stem Cell Research Laboratory of Gulou Hospital Affiliated to Nanjing University School of Medicine,Nanjing 210008,Jiangsu Province,China;Clinical Stem Cell Research Laboratory,Nanjing Gulou Hospital,Gulou Clinical School of Nanjing University of Chinese Medicine,Nanjing 210008,Jiangsu Province,China;School of Life Sciences,Nanjing University,Nanjing 210008,Jiangsu Province,China)

机构地区：[1]南京大学医学院附属鼓楼医院临床干细胞研究室,南京大学生命科学学院,江苏省南京市210008 [2]南京大学医学院附属鼓楼医院临床干细胞研究室,江苏省南京市210008 [3]临床干细胞研究室,南京鼓楼医院,南京中医药大学鼓楼临床医学院,江苏省南京市210008 [4]南京大学生命科学学院,江苏省南京市210008

出　　处：《中国组织工程研究》2025年第23期4878-4887,共10页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(82070459,82270701),项目负责人:王斌。

摘　　要：背景:间充质干细胞来源广泛、易体外增殖,并可分泌一系列免疫调节因子抑制炎症、促进组织修复再生,广泛应用于各种疾病治疗研究。但是对于多种疾病,间充质干细胞的治疗效果有限。针对疾病特定发病机制或者干预靶点进行工程化修饰间充质干细胞是未来细胞治疗的重要发展方向。目的:白细胞介素10是一种典型的抗炎细胞因子,有助于调节免疫反应并诱导巨噬细胞向抗炎表型极化,该研究探讨白细胞介素10基因工程化修饰人脐带间充质干细胞对炎症性肠病的治疗效果。方法:通过电转染建立稳定过表达人白细胞介素10基因的人脐带间充质干细胞,并基于细胞治疗产品标准筛选出临床级细胞。给予C57BL/6J小鼠自由饮用5%葡聚糖硫酸钠盐水溶液建立急性结肠炎模型,分别在造模前1 d(尾静脉途径)与造模后第4天(腹腔途径)注射空质粒转染的人脐带间充质干细胞或过表达人白细胞介素10基因的人脐带间充质干细胞(1×10~6个/只)。在造模后第6天取结肠组织进行苏木精-伊红染色评估组织学变化,免疫荧光染色检测增殖细胞核抗原与CD31的表达。结果与结论:构建的稳定过表达白细胞介素10的工程化人脐带间充质干细胞,满足临床级人脐带间充质干细胞质量标准;人脐带间充质干细胞对小鼠急性结肠炎具有修复效果,过表达白细胞介素10基因的人脐带间充质干细胞的治疗效果更加优效,更显著地抑制急性结肠炎小鼠的体质量下降(P<0.05)、结肠缩短(P<0.05)以及结肠组织损伤(P<0.05);过表达白细胞介素10基因人脐带间充质干细胞组结肠组织切片中增殖细胞核抗原阳性细胞与CD31阳性细胞显著多于人脐带间充质干细胞组,表明过表达白细胞介素10基因的人脐带间充质干细胞可能通过显著促进肠组织细胞增殖和血管再生进而促进结肠组织修复。BACKGROUND:Mesenchymal stem cells have been widely used in the treatment of various diseases due to their wide range of sources,their ease of proliferation in vitro and their ability to secrete a range of immunomodulatory factors to suppress inflammation and promote tissue repair and regeneration.However,in the treatment of many diseases,the therapeutic effect is limited.The effort to engineer and modify mesenchymal stem cells for specific disease pathogenesis or intervention targets is an important development for cell therapy in the future.OBJECTIVE:Interleukin-10 is a typical anti-inflammatory cytokine that helps to modulate the immune response and induces macrophage polarization towards an anti-inflammatory phenotype.This study investigated the therapeutic effect of interleukin-10 engineered human umbilical cord mesenchymal stem cells on inflammatory bowel disease.METHODS:Human umbilical cord mesenchymal stem cells stably overexpressing interleukin-10 were established by electro-transfection,and screened for clinical-grade cells based on the cell therapy product criteria.C57BL/6J mice were given 5%aqueous dextran sulfate sodium ad libitum to establish a model of acute colitis.Empty plasmid-transfected human umbilical cord mesenchymal stem cells or interleukin-10-human umbilical cord mesenchymal stem cells(1×106 cells/each mouse)were injected on day 1 before modeling(tail vein injection)and day 4(intraperitoneal injection)after modeling,respectively.On day 6 after modeling,colon tissue sections were taken for hematoxylin-eosin staining to assess histological changes.Immunofluorescence staining was performed to detect the expression of proliferating cell nuclear antigen and CD31.RESULTS AND CONCLUSION:The engineered human umbilical cord mesenchymal stem cells stably overexpressing interleukin-10 were constructed,and met the quality standard of clinical-grade human umbilical cord mesenchymal stem cells.Human umbilical cord mesenchymal stem cells could repair acute colitis in mice.The therapeutic effect of inter

关 键 词：白细胞介素10 人脐带间充质干细胞 工程化修饰间充质干细胞 炎症性肠病 再生修复 细胞增殖 肠再生 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R574