间充质干细胞移植对降植烷诱导狼疮小鼠肺泡出血的影响及可能机制  

The effect and possible mechanism of mesenchymal stem cell transplantation on pristane induced pulmonary alveolar hemorrhage in lupus-prone mice

作　　者：张卓亚[1] 陈晨 黄赛赛[1] 姜波[1] ZHANG Zhuoya;CHEN Chen;HUANG Saisai;JIANG Bo(Department of Rheumatology and Immunology,Nanjing Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University,Nanjing Jiangsu 210008,China;Department of Nutrition,Nanjing Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University,Nanjing Jiangsu 210008,China)

机构地区：[1]南京大学医学院附属鼓楼医院风湿免疫科,江苏南京210008 [2]南京大学医学院附属鼓楼医院临床营养科,江苏南京210008

出　　处：《江苏大学学报（医学版）》2024年第3期185-189,196,共6页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81901644);南京市卫生科技发展专项资金项目(YKK19051)。

摘　　要：目的:探讨间充质干细胞(mesenchymal stem cells,MSCs)移植对降植烷诱导狼疮小鼠肺泡出血的作用及可能机制。方法:10周龄雌性C57BL/6(B6)小鼠20只腹腔注射0.5 mL降植烷,3只同周龄雌性B6小鼠腹腔注射PBS作为正常对照组。1周后将20只注射降植烷小鼠随机分为MSC组和PBS组,分别给予尾静脉注射1×10^(6)人脐带来源的MSCs和PBS;观察1周后,计算造模小鼠生存率并称量小鼠体重,处死所有小鼠,取出肺脏,观察肺脏大体及肺脏HE染色评估造模小鼠肺出血严重程度。酶消化法制备获取3组小鼠肺脏单细胞悬液,流式细胞术检测3组小鼠肺部CD4^(+)、CD8^(+)T淋巴细胞,CD19^(+)B淋巴细胞,CD11b^(+)Gr1^(+)中性粒细胞,F4/80^(+)肺间质巨噬细胞的绝对数以及抗炎型CD206^(+)肺间质巨噬细胞的百分率。结果:MSC组小鼠生存率高于PBS组,但差异无统计学意义(P=0.07);体重明显高于PBS组小鼠(P<0.01);MSC组小鼠完全弥漫性肺泡出血(diffuse alveolar hemorrhage,DAH)及部分DAH的发生率低于PBS组。与PBS组相比,MSC组肺脏单细胞总数显著减少(P<0.01),肺脏CD4^(+)T细胞数量有下降趋势,而CD8^(+)T细胞及CD19^(+)B细胞的数量则明显下降(P<0.05),肺脏CD11b^(+)Gr1^(+)中性粒细胞及F4/80^(+)巨噬细胞的绝对数也均显著降低(P<0.01和P<0.05);此外,肺出血小鼠CD206^(+)抗炎型巨噬细胞的百分率显著降低(P<0.01),而MSC移植显著提高CD206^(+)抗炎型巨噬细胞的百分率(P<0.01)。结论:MSC移植可显著降低狼疮小鼠肺泡出血的发生率,其机制可能是对肺脏免疫细胞的调控及诱导巨噬细胞CD206抗炎表型的产生。Objective:To investigate the effect of mesenchymal stem cells(MSC)transplantation on pristane induced pulmonary alveolar hemorrhage in lupus mice and explore the possible mechanism.Methods:Twenty female C57BL/6(B6)mice at 10 weeks of age were injected with 0.5 mL pristane intraperitoneally and 3 control B6 mice were injected with the same volume of PBS.After one week,mice injected with pristane were randomly divided into two groups and intravenously injected with 1×10^(6) human umbilical cord-derived MSCs or PBS,respectively.After another week of observation,the survival rate and body weight of the two groups were recorded and all mice were sacrificed.The appearance of the lungs was observed and hematoxylin and eosin(HE)staining was performed to assess the severity of pulmonary hemorrhage.Lung single-cell suspensions were prepared by enzymatic digestion.CD4^(+)and CD8^(+)T lymphocytes,CD19^(+)B lymphocytes,CD11b^(+)Gr1^(+)neutrophils,F4/80^(+)macrophages and CD206^(+)macrophages in lungs were detected by flow cytometry.Results:The survival rate of mice in the MSC group was higher than that of the PBS group,but the difference was not statistically significant(P=0.07),and the average body weight in MSC mice was significantly increased(P<0.01)than that in PBS group.The incidences of complete diffuse alveolar hemorrhage and partial diffuse alveolar hemorrhage in the MSC group were lower than that in the PBS group.Compared with the PBS mice,the total number of lung cells in the MSC group was significantly reduced(P<0.01)than that in the PBS group.The number of lung CD4^(+)T cells in the MSC group showed a decreased tendency,while the numbers of CD8^(+)T cells and CD19^(+)B cells were significantly decreased(P<0.05)in the MSC group than that in the PBS group.The absolute numbers of CD11b^(+)Gr1^(+)neutrophils and F4/80^(+)macrophages in lungs of MSC treated mice were also markedly reduced(P<0.01 and P<0.05).In addition,the percentage of CD206^(+)anti-inflammatory macrophages in lungs of pulmonary hemorrhage mice was s

关 键 词：间充质干细胞 系统性红斑狼疮 肺泡出血 巨噬细胞 CD206 

分 类 号：R593.243[医药卫生—内科学]