山西白求恩医院,太原 030032
R-33
Shanxi Bethune Hospital, Taiyuan 030032, China
目的 探讨 β-氨基丙腈(β-aminopropionitrile,BAPN)联合血管紧张素Ⅱ( angiotensin II,Ang II)诱导主动脉夹层小鼠主动脉中的巨噬细胞表型变化的情况。 方法 通过主动脉夹层相关的转录组芯片,寻找相关的核心基因及相关的生物学功能及信号通路情况;进一步利用 CIBERSORT 分析主动脉中各型细胞的比例;将清洁级雄性 C57BL/ 6 小鼠 40 只,随机分为 BAPN 腹腔注射组,Ang II 腹腔注射组,BAPN 和 Ang II 联合注射组,生理盐水腹腔注射组,每组 10 只。 BAPN 注射剂量:100 mg / (kg·d),Ang II 注射剂量:4 mg / ( kg·d),BAPN 和 Ang II 联合剂 量为 100 mg / (kg·d)和 4 mg / (kg·d)。 每日按实验方法对不同组别小鼠给予药物处理,给药 2 周,构建小鼠主动脉夹层模型;随后利用免疫荧光、蛋白质免疫印迹和酶联免疫吸附实验研究主动脉夹层中 M1 和 M2 型巨噬细胞的 表型变化。 结果 通过分析 GSE147026 发现主动脉夹层相关的差异基因共 930 个,生物学功能主要涉及白细胞与细胞的粘附、细胞外结构组织、巨噬细胞激活、细胞外基质组织、白细胞迁移;分子功能主要集中在膜微区、膜筏、粘着斑、细胞-底物结、胶原蛋白细胞外基质;生物学过程主要集中在整联蛋白结合、Toll 样受体结合、清道夫受体活性、受体活性、细胞外基质结构成分等。 使用 CIBERSORT 分析显示,幼稚 B 细胞、M1 巨噬细胞在主动脉夹层组织中表现出更高的浸润水平,活化 NK 细胞在正常主动脉组织表现出更高的浸润水平,M0 和 M2 巨噬细胞在主动脉 夹层组织中表现出较高的浸润水平,但差异无统计学意义,余其他细胞类型在正常主动脉和主动脉夹层组织中没 有显著差异。 iNOS、CD206、IL-6 和 IL-10 蛋白表达相较于正常组而言,在 BAPN+Ang II 组最高,BAPN 次之,Ang II 最少。 结论 主动脉夹层小鼠主动脉组织中均有 M1 型和 M2 型巨噬细胞的蓄积,其中 BAPN+Ang II 组蓄积量最多,BAPN 次之,Ang II 最少。
Objective To investigate the effect of BAPN combined with AngII ( BAPN + Ang II) on the phenotypic changes of macrophages in the aorta of mice with aortic dissection. Methods Searching for relevant core genes and related biological functions and signaling pathway profiles through transcriptome microarrays related to aortic coarctation. We then applied CIBERSORT to analyze the proportion of various types of cells in the aorta. Last, we conducted immunofluorescence, Western blot, and enzyme-linked immunosorbent assays to study the phenotypic changes of M1 and M2 macrophages in aortic dissection. Results GSE147026 found a total of 930 differential genes related to aortic dissection. The biological functions of these genes involve mainly the adhesion of leukocytes to cells, extracellular structure organization, macrophage activation, extracellular matrix organization, and leukocyte migration. Their molecular functions are concentrated mainly in the membrane: microdomains, membrane rafts, adhesion spots, cell-substrate junctions, and collagen extracellular matrix, and their biological processes focus mainly on integrin binding, Toll-like receptor binding, scavenger receptor activity, receptor activity, and extracellular matrix structure components. The CIBERSORT analysis revealed that naive B cells and M1 macrophages exhibited higher infiltration levels in aortic dissection tissues, activated NK cells displayed higher infiltration levels in normal aortic tissues, and M0 and M2 had higher infiltration levels in aortic dissection tissues; however, these differences did not reach statistical significance. No other cell types presented a significantly different level of abundance between normal aorta and aortic dissection. Compared with the normal group, the protein expression levels of iNOS, CD206, IL-6 and IL-10 were the highest in the BAPN+Ang II group, followed by the BAPN group, with the Ang II group having the lowest levels. Conclusions Both M1 and M2 macrophages accumulate in the aorta of aortic dissection mice; their relative accumulation levels in our groups were as follows: Among them, the BAPN + AngII group had the largest accumulation, followed by BAPN and the least AngII.
张立魁,杨灵波,王学宁.主动脉夹层中巨噬细胞表型变化研究[J].中国比较医学杂志,2022,32(8):62~70.
复制