| 摘要: |
| 目的 探讨3'-羟基染料木素对3T3-L1前脂肪细胞增殖、分化的影响及其机制。 方法 培养3T3-L1细胞,用3'-羟基染料木素干预,用四甲基偶氮唑盐(MTT)法和细胞计数法检测其对细胞增殖的影响,用5-溴脱氧尿嘧啶(bromodeoxyuridine,BrdU)掺入法检测其对前脂肪细胞内DNA合成的影响;用油红O染色法检测其对前脂肪细胞分化过程中脂质的堆积的影响;采用RT-PCR和蛋白免疫印迹技术检测过氧化物酶体增殖子活化受体γ(peroxisome proliferator activated receptor γ, PPARγ)和CAAT/增强子结合蛋白α(CAAT/enhancer binding protein α, C/EBPα)的mRNA以及蛋白的表达情况。 结果 用10~50 μmol/L 3'-羟基染料木素分别处理前脂肪细胞24、48和72 h,能促进前脂肪细胞增殖(P<0.05或P<0.01),且呈一定的量效关系;3'-羟基染料木素能浓度依赖性地促进前脂肪细胞内DNA合成(P<0.05);3'-羟基染料木素与噻唑烷二酮类药物罗格列酮(rosiglitazone,ROZ)相似,在10和50 μmol/L 浓度下能使前脂肪细胞的胞浆中出现较大量的脂滴(P<0.05或P<0.01);3'-羟基染料木素处理组中PPARγ和C/EBPα的mRNA与蛋白的表达增加(P<0.01)。 结论 3'-羟基染料木素能够促进前脂肪细胞的增殖与分化,增加脂肪细胞分化过程中脂质的堆积,其机制可能与其促进PPARγ、和C/EBPα的mRNA与蛋白表达有关。 |
| 关键词: 3'-羟基染料木素 前脂肪细胞 细胞增殖 细胞分化 PPARγ |
| DOI：10.3724/SP.J.1008.2011.01181 |
| 投稿时间：2011-06-21修订日期：2011-10-18 |
| 基金项目:上海市卫生局青年科研基金(2009Y069). |
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| Effects of 3'-hydroxygenistein on proliferation and differentiation of 3T3-L1 preadipocytes and the underlying mechanisms |
| CHEN Li1,2*,SANG Hao2,LI Jue2 |
(1. Department of Pharmacy, Central Hospital of Xuhui District, Shanghai 200031, China;
2. Department of Preventive Medicine, Tongji University School of Medicine, Shanghai 200092, China
*Corresponding author.) |
| Abstract: |
| Objective To explore the effects of 3'-hydroxygenistein on the proliferation and differentiation of 3T3-L1 preadipocytes and to elucidate the related mechanism. Methods 3T3-L1 preadipocytes were cultured and treated with different dosages of 3'-hydroxygenistein. Cell proliferation was analyzed by counting cell numbers and MTT assay; DNA synthesis of 3T3-L1 was investigated by bromodeoxyuridine (BrdU) incorporation assay; the degree of preadipocytes differentiation was evaluated by Oil red O staining; and the expressions of peroxisome proliferation activated receptor γ (PPARγ) and CAAT/enhancer binding protein (C/EBPα) were detected at mRNA and protein level by real-time PCR and Western blotting analysis, respectively. Results Pretreatment with 3'-hydroxygenistein (10-50 μmol/L) for 24, 48 and 72 h markedly promoted 3T3-L1 preadipocyte proliferation and differentiation in a dose-effect manner (P<0.05, P<0.01). It also significantly facilitated the DNA synthesis in 3T3-L1 preadipocytes in a dose-dependent manner (P<0.05). Similar to rosiglitazone (ROZ), 3'-hydroxygenistein (at 10 or 50 μmol/L) also resulted in more lipid droplets in 3T3-L1 preadipocytes (P<0.05, P<0.01). Furthermore, 3'-hydroxygenistein greatly increased the mRNA and protein expression of PPARγ and C/EBPα in 3T3-L1 preadipocytes (P<0.01). Conclusion 3'-hydroxygenistein can promote the proliferation and DNA synthesis of 3T3-L1 preadipocytes; it can also enhance the accumulation of lipid drops and increase the terminal differentiation of preadipocyts, which might be associated with its ability to increase the expression of PPARγ and C/EBPα. |
| Key words: 3'-hydroxygenistein preadipocyte cell proliferation cell differentiation PPAR gamma |
