树大网膜前脂肪细胞原代培养及诱导分化

目的探讨树大网膜前脂肪细胞原代培养的方法,优化其诱导分化的方法。方法采用酶消化的方法对树大网膜组织进行原代培养,待其传至第三代时,用含0.5mmoL/L3-异丁基-1-甲基黄嘌呤、10μmol/L胰岛素、0.2mmoL/L吲哚美辛、1μmoL/L地塞米松的高糖DMEM诱导配方对树大网膜前脂肪细胞进行成脂诱导,14d后见到细胞内聚集脂滴,对其进行油红O染色。结果树大网膜脂肪细胞在体外成功培养,细胞形态良好,呈梭形。诱导后的脂肪细胞第3d出现少数出现小脂滴,14d左右,大部分细胞胞质中出现圆而大的脂滴。结论树大网膜脂肪组织可在体外进行培养,稳定传代并能诱导分化为成熟的脂肪细胞。     

瘦素对人前脂肪细胞增殖及分化的影响

目的 观察瘦索在体外对人前脂肪细胞增殖和分化的影响,探讨瘦素调节肥胖发生的可能机制.方法 分离并体外培养人腹部皮下前脂肪细胞.采用3-（4,5-二甲基噻唑-2）-2,5-二苯基四氮唑溴盐（MTT）比色法、细胞计数法、油红O染色提取法及逆转录-聚合酶链式反应（RT-PCR）方法 分析不同浓度（0～1 000 ng/ml）瘦素对人前脂肪细胞增殖、脂质积聚及分化转录因子γ2过氧化物酶体增殖物激活受体（PPAR-γ）、CCAAT增强子α结合蛋白（C/EBP-α）mRNA表达的影响.结果 高浓度（1 000 ng/ml）瘦素能够促进人前脂肪细胞的增殖、脂质积聚以及PPAR-γ2、C/EBP-α mRNA表达（P〈0.05）.低浓度（10 ng/ml）和中浓度（100 ng/ml）瘦素对人前脂肪细胞的增殖及脂质积聚没有明显的促进作用（P〉0.05）.结论 在体外超生理浓度的瘦素能够促进前脂肪细胞的增殖和分化,提示瘦素抵抗、血清高瘦素浓度等病理状态时,瘦素可能促进前脂肪细胞的增殖及分化,影响肥胖发生.     

染料木苷对3T3-L1脂肪细胞分化以及AMPK磷酸化的影响

目的观察染料木苷(Genistin,GIN)对3T3-L1前脂肪细胞细胞分化以及分化过程中腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)磷酸化程度的影响。方法用激素鸡尾酒法诱导3T3-L1前脂肪细胞分化,用MTT法观察GIN对细胞生存率的影响,以油红O染色法镜下观察3T3-L1前脂肪细胞分化的情况,并通过比色分析法检测GIN对细胞内脂滴堆积量的影响;用Western blotting方法分别检测对照组和给药组细胞分化过程中细胞中AMPK,P-AMPK(Thr172)蛋白表达量,借以计算AMPK磷酸化比率。结果分化诱导后,对照组脂肪细胞呈典型的成熟脂肪细胞表型,形成"戒指环"结构,经100μM GIN干预后,脂滴数量减少,体积减小,通过比色分析法量化后显示100μM GIN可减少胞内脂质的堆积(P<0.05)。与对照组相比,给药组从第4天开始上调p-AMPK(Thr172)/AMPK比率,并持续到第10天(P<0.05)。结论 GIN可通过激活AMPK抑制3T3-L1脂肪细胞分化。     

Equisetin is an anti-obesity candidate through targeting 11β-HSD1

Obesity is increasingly prevalent globally, searching for therapeutic agents acting on adipose tissue is of great importance. Equisetin (EQST), a meroterpenoid isolated from a marine sponge-derived fungus, has been reported to display antibacterial and antiviral activities. Here, we revealed that EQST displayed anti-obesity effects acting on adipose tissue through inhibiting adipogenesis in vitro and attenuating HFD-induced obesity in mice, doing so without affecting food intake, blood pressure or heart rate. We demonstrated that EQST inhibited the enzyme activity of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), a therapeutic target of obesity in adipose tissue. Anti-obesity properties of EQST were all offset by applying excessive 11β-HSD1's substrates and 11β-HSD1 inhibition through knockdown in vitro or 11β-HSD1 knockout in vivo. In the 11β-HSD1 bypass model constructed by adding excess 11β-HSD1 products, EQST's anti-obesity effects disappeared. Furthermore, EQST directly bond to 11β-HSD1 protein and presented remarkable better intensity on 11β-HSD1 inhibition and better efficacy on anti-obesity than known 11β-HSD1 inhibitor. Therefore, EQST can be developed into anti-obesity candidate compound, and this study may provide more clues for developing higher effective 11β-HSD1 inhibitors.