小鼠M1和M2巨噬细胞肝内移植模型的建立  

Establishing a mouse model of M1/M2 - polarized macrophages intrahepatic transplantation

作　　者：孙航 王恺[1] 谢浩荣 陈刚 林辛辛 李川江[1] 周杰[1] Sun Hang;Wang Kai;Xie Haorong;Chen Gang;Lin Xinxin;Li Chuanjiang;Zhou Jie(Department of Hepatobiliary Surgery, Nanfang Hospital, Southern Medical University, Guangzhou , 510515, China （Sun H, Wang K, Xie HR, Chen G, Li C J, Zhou J;The First Clinical College, Southern Medical University, Guangzhou, 510515, China （L in XX)

机构地区：[1]南方医科大学南方医院肝胆外科,广州510515 [2]南方医科大学第一临床医学院,广州510515

出　　处：《中华实验外科杂志》2018年第7期1354-1356,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81600462）;广州市科技计划项目（201508020262-1）;南方医科大学南方医院杰出青年培育计划（2016006）

摘　　要：目的建立小鼠M1／M2极化巨噬细胞肝内移植模型。方法分离、培养小鼠骨髓来源的巨噬细胞（BMDM），并将其进行荧光标记、极化，获得M1型和M2型巨噬细胞，未极化细胞标记为M0，将3种细胞用经门静脉注射8×10^5个细胞至同种异体小鼠肝内，设置磷酸盐缓冲液（PBS）注射组为对照组，于术后24、72h取肝组织采用荧光显微镜检测荧光标记的BMDM，并检测M1、M2BMDM相关基因mRNA表达。结果BMDM经分离、培养、分化获得F4／80阳性巨噬细胞[占（97．60±1．25）％]，并经转染、极化成功获得荧光标记的M1、M2BMDM，移植至同种异体小鼠肝内24、72h后，肝组织内均可检测到荧光标记的M0、M1、M2BMDM，且肝组织中M1、M2BMDM相关基因mRNA表达显著上调。结论成功建立分离极化后的小鼠巨噬细胞肝内移植模型。Objective To establish a M1/M2 -polarized macrophages intrahepatic transplantation model in mice. Methods Bone marrow -derived macrophages （BMDMs） were harvested. After carboxyfluorescein （FAM） - labeled small interfering RNA （siRNA） transfection, the transfected BMDM were incubated to induce M1 and M2 polarization, respectively. The BMDM were not induced marked M0. Then the 8 × 10^5 polarized BMDM were transfused to mouse liver through portal vein injection. The mouse liver injected phosphate buffer solution （PBS） were assigned to control group. FAM positive BMDM ware examined in liver tissues under fluorescence microscopy, and the mRNA levels of polarized - macrophages signature genes were measured by real -time quantitative polymerase chain reaction （real -time PCR） assays. Results Bone marrow cells were differentiated into 174/80 positive macrophages （97.60 ± 1.25 ）%. After transfection, FAM positive M1 and M2 BMDM were successfully obtained. M1 and M2 signature genes were highly expressed in M1 and M2 BMDM, respectively. 24 hours and 72 hours after MI and M2 BMDM portal vein injection, the FAM positive M0, M1 and M2 BMDM could be detected in the liver tissues of recipient mice. Moreover, M1 and M2 signature genes were significantly upregulated in M1 and M2 BMDM injected liver, respectively. Conclusion This model provides an ideal animal model to investigate the function of macrophage polarization in liver disease.

关 键 词：巨噬细胞极化 肝内移植 小鼠模型 

分 类 号：R-332[医药卫生] R575