中文摘要：

目的探讨人皮下、网膜脂肪组织来源的前脂肪细胞形态、功能的差异，观察不同浓度甘精胰岛素对其增殖、分化的影响。方法胶原酶分离法原代培养人皮下、网膜前脂肪细胞，倒置显微镜下观察不同来源的人前脂肪细胞在形态方面的差异，不同浓度甘精胰岛素（20、200、500、100（3、1500nmol／L）干预其增殖及分化过程，四甲基偶氮唑盐（MTT）法检测二者增殖的差异，实时定量PCR检测脂肪分化相关基因表达的差异。结果（1）可从人皮下、网膜脂肪组织分离出前脂肪细胞，并在体外扩增，皮下前脂肪细胞更细长，容易增殖，而网膜前脂肪细胞呈多角形，易于老化。（2）MTT法结果显示甘精胰岛素抑制网膜前脂肪细胞的体外增殖，且其抑制作用具有剂量依赖性，1000nmol／L干预72h后，与未加胰岛素组相比，网膜前脂肪细胞的增殖明显低[吸光度（A）值：0．144±0．021比0．267±0．040，P〈0．01]；皮下前脂肪细胞作用不明显（A值：0．305±0．045比0．350±0．037，P〉0．05）。（3）500nmol／L的胰岛素浓度是人前脂肪细胞分化的适宜浓度，PCR结果显示该浓度时，对于皮下前脂肪细胞，脂肪分化的标志基因过氧化物酶增殖物激活受体（PPAR）γ（F=31．31，P〈0．01）和CCAAT／增强子结合蛋白0l（C／EBP（x）mRNA表达最高（F=9．86，P〈0．05），前脂肪细胞的标志基因Pref-1表达最低（皮下F=68．95，P〈0．01；网膜F=30．38，P〈0．01），但是胰岛素浓度对网膜脂肪细胞PPARγ、C／EBPαmRNA影响不大（均P〉0．05）。结论甘精胰岛素可抑制人网膜前脂肪细胞的增殖，促进皮下、网膜前脂肪细胞的分化。

英文摘要：

Objective To compare the morphological and functional differences of human primary preadipoeytes from different fat depots and explore the effects of insulin glargine on their proliferation and differentiation. Methods Primary preadipocytes isolated from human subcutaneous and omental adipose tissue by collagenase I were passaged in vitro. Inverted phase contrast microscope was used to observe the morphological differences of two kinds of preadipocytes. Then two kinds of preadipocytes were cultured or induced to differentiation with different doses of insulin glargine. The methyl thiazolyl tetrazolium （MTT） assay was used to detect their proliferative differences. Reverse transcription-polymerase chain reaction （ RT- PCR） was used to observe the effects of insulin on adipogenie gene expression. Results （1） Both preadipocytes could be successfully cultured from adipose tissue and amplified in vitro. Subcutaneous preadipocytes were more slender and proliferated more quickly while omental preadipocytes were polygonal and aged easily. （2） MTT results showed that insulin glargine could inhibit the proliferation of omental preadipocytes in a dose-dependent fashion. After 72 h incubation, compared with negative control, the absorbance （A） value of 1000 nmol/L insulin glargine group decreased greatly （0. 144 ±0. 021 vs 0. 267 ± 0. 040, P 〈 0. 01 ）. But it had no effect on subcutaneous preadipocytes （0. 305 ±0. 045 vs 0. 350 ±0. 037, P 〉 0. 05 ）. （ 3 ） Insulin at 500 nmol/L was a suitable concentration for inducing differentiation. RT-PCR analysis showed that, for subcutaneous adipocytes, adipogenic genes such as peroxisome proliferator- activated receptor gamma （PPARγ） （F = 31.31, P 〈 0. 01 ）and CCAAT enhancer binding protein α （C/EBPct） （F = 9.86, P 〈 0. 05 ）had the highest mRNA expression while preadipoeytic gene Pref-1 had the lowest expression at this concentration. But insulin dose had no obvious effect on PPARγ or C/EBPα mRNA （ P 〉 0. 05 ） for om