ox-LDL诱导后SHP-2缺陷骨髓巨噬细胞脂肪沉积、吞噬功能及脂肪代谢基因表达观察  

Fat deposition, phagocytosis and expression of fat metabolism-related genes in myeloid macrophages of SHP-2 gene deficient mice induced by ox-LDL

作　　者：梁雪雷 马倩[2] 李新新[2] 谭鹤[2] 张雪 帖彦清[2] LIANG Xuelei;MA Qian;LI Xinxin;TAN He;ZHANG Xue;TIE Yanqing(Hebei North University,Zhangjiakou 075000,China)

机构地区：[1]河北北方学院,河北张家口075000 [2]河北省人民医院

出　　处：《山东医药》2019年第32期20-24,共5页Shandong Medical Journal

基　　金：河北省政府资助临床医学优秀人才培养和基础课题研究项目计划(361003)

摘　　要：目的观察氧化型低密度脂蛋白(ox-LDL)诱导后SHP-2基因缺陷小鼠骨髓来源巨噬细胞(BMDM)脂肪沉积、吞噬功能及脂肪代谢相关基因表达的变化。方法以Lyz-2-Cre小鼠和SHP-2^flox/flox小鼠杂交纯化后,繁育骨髓单核-巨噬细胞条件性SHP-2缺陷(SHP-2 MφCKO)小鼠(基因型SHP-2^flox/flox)为实验组,以同窝野生型小鼠(基因型SHP-2^+/+)为对照组。分离两组BMDM并给予ox-LDL刺激48 h后,透射电镜下观察BMDM脂滴沉积情况,用荧光显微镜照相,以红色荧光数量表示泡沫细胞形成情况;激光共聚焦显微镜观察BMDM对单分散荧光微球的吞噬情况,以绿色荧光强度表示BMDM的吞噬功能。ox-LDL刺激两组细胞24 h后采用Q-PCR法检测脂肪代谢相关基因CD36、TLR-4、SREBP-1c、SREBP-2、ABCA1、ABCG1 mRNA。结果繁育出Cre^-小鼠36只,SHP-2 MφCKO小鼠53只,SHP-2^+/+小鼠61只,SHP-2^flox/+小鼠69只。实验组BMDM细胞质内红色和绿色荧光含量均高于对照组(P均<0.05)。实验组BMDM中CD36、TLR-4、SREBP-1c、SREBP-2 mRNA相对表达量高于对照组(P均<0.05)。结论SHP-2基因缺陷小鼠BMDM脂肪沉积加剧,细胞吞噬功能增强,脂肪代谢相关基因表达上调。SHP-2基因缺陷加速泡沫细胞形成可能与增强ox-LDL内化进入巨噬细胞、增强胆固醇或脂质的转运有关。Objective To observe the changes of fat deposition,phagocytosis and expression of fat metabolism-related genes in bone marrow-derived macrophages(BMDM)of protein tyrosine phosphatase(SHP-2)gene deficient mice induced by oxidized-low density lipoprotein(ox-LDL).Methods After hybrid purification of Lyz-2-cre mice and SHP-2 flox/flox mice,bone marrow monocyte-macrophage conditioned SHP-2 deficiency(shp-2 MφCKO)mice(shp-2^flox/flox)were bred as the experimental group,and wild type mice(shp-2^+/+)were used as the control group.BMDM of the two groups were separated and stimulated with ox-LDL for 48 h.The deposition of BMDM lipid droplets was observed under transmission electron microscope and photographed by fluorescence microscope.The formation of foam cells was represented by red fluorescence quantity.The phagocytosis of monodisperse fluorescent microspheres by BMDM was observed by laser confocal microscope,and the phagocytic function of BMDM was expressed by green fluorescence intensity.The fat metabolism-related genes CD36,TLR-4,SREBP-1c,SREBP-2,ABCA1,and ABCG1 mRNA was detected by Q-PCR at 24 h after stimulation with ox-LDL.Results Thirty-six Cre-mice,53 SHP-2 MφCKO mice,61 SHP-2^+/+mice,and 69 SHP-2 flox/+mice were bred.The content of red fluorescence in the cytoplasm of BMDM in the experimental group was higher than that in the control group(P<0.05).The green fluorescence intensity in the cytoplasm of BMDM in the experimental group was higher than that in the control group(P<0.05).The relative expression of CD36,TLR-4,SREBP-1c,and SREBP-2 mRNA in BMDM of the experimental group was higher than that of control group(P<0.05).Conclusions The fat deposition of BMDM in SHP-2 gene deficient mice is aggravated,the phagocytosis function of cells is enhanced,and the expression of fat metabolism-related genes is up-regulated.The acceleration of foam cell formation by SHP-2 gene deficiency may be related to the enhancement of ox-LDL internalization into macrophages and the enhancement of cholesterol or lipid transport.

关 键 词：动脉粥样硬化 蛋白酪氨酸磷酸酶 巨噬细胞 泡沫细胞 脂肪代谢 

分 类 号：R541.4[医药卫生—心血管疾病]