目的探讨C57BL/6J小鼠棕色脂肪成脂分化功能随着年龄增加的变化情况。方法选取0周(初生)、4周、8周、12周C57BL/6J雌雄小鼠各4只,分别取其肩胛部的棕色脂肪组织。采用免疫组织化学法来鉴别其棕色脂肪,之后提取棕色脂肪的总RNA,并对总RNA的质量进行鉴定。通过实时荧光定量PCR法检测0周、4周、8周、12周C57BL/6J小鼠棕色脂肪成脂分化相关基因PPARα、C/EBPα、PGC-1α、PPARγ、FOXC2、BMP7的表达水平。结果解耦联蛋白-1(UCPl)免疫组织化学显示阳性着色较深的为C57BL/6J小鼠棕色脂肪组织,实时荧光定量PCR结果显示成脂分化基因的表达量随着小鼠年龄增加逐渐降低,各时间点之间差异有统计学意义[PPARα(F=11.96,P〈0.0001)、C/EBPα(F=9.39,P〈0.0001)、PGC-1α(F=17.21,P〈0.0001)、PPARγ(F=13.11,P〈0.0001)、FOXC2(F=12.23,P〈0.0001)、BMP7(F=16.44,P〈0.0001)]。结论小鼠棕色脂肪成脂分化能力随着小鼠年龄增长逐渐减退,活性逐渐降低。但其功能的调控因素尚需进一步研究探索。
Objective To explore the status of C57BL/6J mouse brown fat adipogenic differentiation function with aging. Methods C57BL/6J female and male mice at the ages of 0 -week (newborn) ,4 -week, 8 -week, 12 - week old were selected from the same brood, brown adipose tissue was obstained from their interscapular region, and the brown adipose was identified by using immunohistochemical markers. Then the total RNA was extracted from the brown adipose and quality identification was determined at the same time. The expression levels of the related genes (PPARα,C/EBPα, PGC- 1α, PPART, FOXC2, BMP7) induced by brown adipose adipogenic differentiation were detected by quantitative real - time PCR in 0 - week,4 - week,8 - week, 12 - week mice. Results Uncoupling protein - 1 ( UCP1 ) immunohistochemical data indicated that positive deep - colour substance was brown adipose tissue. Quanti- tative Real -time PCR also indicated that the expression volume of adipogenesis gene gradually reduced with aging, and there were significant differences at the different time points [ PPARα ( F = 11.96, P 〈 0.000 1 ), C/EBPα ( F = 9.39, P 〈 0. 000 1 ) ,PGC -1α( F = 17.21 ,P 〈0.000 1 ) ,PPART( F = 13.11 ,P 〈0.000 1 ) ,FOXC2( F = 12.23 ,P 〈 0.000 1 ), BMP7( F = 16.44 ,P 〈0.000 1 )]. Conclusions The adipogenic differentiation ability and activity of mouse brown adi- pose gradually reduce with aging. But the regulatory factors for its function needs to be further investigated.