三磷腺苷合酶抑制因子1基因敲除对小鼠成纤维细胞中三磷腺苷水平及脂肪细胞分化的影响  

Effect of adenosine triphosphate synthase inhibitory factor 1 gene knockout on ATP level and differentiation of adipocytes in fibroblasts of mice

作　　者：关丫丫 梁银明 王辉 叶建平 GUAN Ya-ya;LIANG Yin-ming;WANG Hui;YE Jian-ping(School of Laboratory Medicine,Xinxiang Medical University,Xinxiang 450003,Henan Province,China;Henan Province Collaborative Innovation Center of Molecular Diagnosis and Laboratorial Medicine,Xinxiang 450003,Henan Province,China;Biomedical Center of Pennington,Louisiana State University,Baton Rouge 70808,Louisiana,American)

机构地区：[1]新乡医学院医学检验学院,河南新乡453003 [2]河南省分子诊断与医学检验技术协同创新中心,河南新乡453003 [3]美国路易斯安那州立大学Pennington生物医学中心,路易斯安那州巴吞鲁日市70808

出　　处：《新乡医学院学报》2018年第8期658-661,共4页Journal of Xinxiang Medical University

基　　金：国家自然科学基金资助项目(编号:81370915);校特聘教授科研启动基金资助项目(编号:200/502008)

摘　　要：目的探讨三磷腺苷合酶抑制因子1(ATPIF1)基因敲除对小鼠成纤维细胞中三磷腺苷(ATP)水平及脂肪细胞分化的影响。方法取5只ATPIF1基因敲除小鼠作为观察组,另取5只C57BL/6小鼠作为对照组。取各组小鼠耳组织,采用Ⅰ型胶原酶和中性蛋白酶Ⅱ的消化作用制备成纤维细胞并用高糖培养基培养,传代1~2次后,待细胞长满且接触抑制3 d后,更换含白色脂肪细胞诱导剂Ⅰ的细胞培养基诱导4 d,再更换含白色脂肪细胞诱导剂Ⅱ的细胞培养基诱导4 d。在诱导前及诱导4、8 d分别采用ATP检测试剂盒和三酰甘油(TG)检测试剂盒检测成纤维细胞中ATP和TG水平。结果诱导前,对照组小鼠原代成纤维细胞中ATP水平明显低于观察组(P<0.05);诱导4、8 d,对照组与观察组小鼠原代成纤维细胞中ATP水平比较差异无统计学意义(P>0.05)。诱导4、8 d,对照组小鼠原代成纤维细胞中TG水平低于观察组(P<0.05)。结论 ATPIF1基因敲除可增加小鼠成纤维细胞中ATP水平,促进成纤维细胞向白色脂肪细胞分化,提高成熟白色脂肪细胞储存TG的能力。Objective To investigate the effect of adenosine triphosphate inhibitory factor 1（ATPIF1） gene knockout on the level of adenosine triphosphate（ATP） and the differentiation of adipocytes in fibroblasts of mice.Methods Five ATPIF1 gene knockout mice were chose as experimental group and five C57 BL/6 mice were chose as control group.Their ears were cut off and digested with collagenase type Ⅰ and neutral protease Ⅱ to obtain the primary fibroblasts from the ears of mice,and the cells were cultured by high sugar medium.The fibroblasts were differentiated into white adipocytes after 1-2 time passages.After the cells were fulled of dish and contacted inhibition for three days,then the cell culture medium was replaced with the prepared white adipocytes reagent Ⅰ for 4 days,then replaced with the prepared white adipocytes reagent Ⅱfor 4 days.The ATP and triacylglycerol（TG） level in fibroblasts were detected by ATP assay kit and TG assay kit before induction and at 4,8 days after induction.Results The ATP level in fibroblasts in the control group was significantly lower than that in the observation group（P〈0.05）.At 4,8 days after induction,there was no significant difference in the ATP level between the two grous（P〈0.05）.The TG level in fibroblasts in the control group was lower than that in the observation group（P〈0.05）.Conclusion The ATPIF1 gene knockout can increase the ATP level in fibroblasts of mice,promote the differentiation of fibroblasts into white adipocytes and improve the ability of storing TG in white adipocytes.

关 键 词：ATP合酶抑制因子1 脂肪细胞分化 三磷腺苷 三酰甘油 

分 类 号：R589.2[医药卫生—内分泌]