高脂饮食小鼠脂肪组织来源DC的形态鉴定及表型分析

Identification and phenotypic analysis of the adipose tissue-derived dendritic cells in high-fat diet mice

有报道高脂饮食(high-fat diet,HFD)小鼠脂肪组织中存在促炎症DC,本研究应用FCM、电镜及实时荧光定量PCR(real-time quantitative PCR,qRT-PCR)等对其进行形态鉴定及表型分析。具体为用FCM检测正常饮食(normal diet,ND)及HFD小鼠脂肪组织中CD11c+DC的百分比;利用电镜观察脂肪组织来源的CD11c+DC的形态;应用FCM检测HFD小鼠脂肪组织中的CD11c+DC表面分子如CD40、CD80、CD86、MHCⅠ及MHCⅡ的表达情况,进而对小鼠脂肪组织中CD11c+DC进行表型分析,同时设立脂多糖(Lipopolysaccharide,LPS)组作为对照。结果显示HFD小鼠脂肪组织来源的CD11c+DC形态典型、具有很长的突触、富含线粒体并且胞质中极少看到溶酶体,其百分比明显高于ND组。经LPS刺激4h后,HFD小鼠脂肪组织来源树突状细胞(adipose tissue dendritic cell,ATDC)的表面分子CD40、CD80、CD86、MHCⅠ及MHCⅡ等表达稳定,说明HFD小鼠ATDC是一群表型稳定的DC。以上结果说明与ND小鼠相比,HFD小鼠脂肪组织中存在着一群百分比相对增高、形态典型且表型稳定的DC。

Some studies have reported that the adipose tissue cells of the high-fat diet(HFD)mice have pro-inflammatory dendritic cells.To evaluate the pattern and phenotypic analysis of the cells by FCM,electron microscope and qRT-PCR.Specifically,FCM was used to detect the percentage of the CD11c+DC in the adipose tissue cells of normal diet(ND)and HFD mice;electron microscope was used to observe the morphology of the CD11c+DC in the adipose tissue cells;the expression of CD11c+DC surface molecules such as CD40,CD80,CD86,MHCⅠand MHCⅡin adipose tissue of HFD mice was detected by FCM.Then the phenotype of CD11c+DC in adipose tissue of mice was analyzed,the experiment of LPS stimulation was set up as the control group.The result showed that the adipose tissue CD11c+DC of HFD mice had a typical pattern,long dendrites,rich mitochondrion in cytoplasm and rare lysosomes.Higher percentage of CD11c+DC was presented in HFD mice adipose tissue than ND.Phenotypic analysis indicated that the expressions of CD40,CD80,CD86,MHCⅠand MHCⅡin HFD adipose tissue derived dendritic cell(ATDC)were stable after 4hours of LPS stimulation.This indicates that the ATDC of HFD mice is a group of phenotypically stable DC.The above results indicate that there is a group of DC with high percentage,typical pattern and stable phenotype in the adipose tissue of HFD mice,compared with ND mice.