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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 acti-vated receptorγ,PPARγ)和CAAT/增强子结合蛋白α(CAAT/enhancer binding proteinα,C/EBPα)的mRNA以及蛋白的表达情况。结果用10～50μmol/L 3′-羟基染料木素分别处理前脂肪细胞24、48和72h,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50μmol/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。
Objective To investigate the effect and mechanism of 3’-hydroxyl genistein on proliferation and differentiation of 3T3-L1 preadipocytes. Methods 3T3-L1 cells were cultured and treated with 3’-hydroxy genistein. The effects of 3-hydroxy genistein on the proliferation of 3T3-L1 cells were detected by MTT assay and cell counting method. The inhibitory effect of 5-bromodeoxyuridine , BrdU) were used to detect the effect on DNA synthesis in preadipocytes. The effect of preadipocyte differentiation on the accumulation of lipids was detected by oil red O staining. RT-PCR and Western blotting MRNA and protein expression of peroxisome proliferator-activated receptor γ (PPARγ) and CAAT / enhancer binding protein α (C / EBPα) were detected. Results Treatment of preadipocytes with 10 ~ 50μmol / L 3’-hydroxy genistein for 24, 48 and 72 hours, respectively, promoted the proliferation of preadipocytes (P <0.05 or P <0.01), and showed a dose-effect relationship. 3 ’-hydroxy genistein could promote DNA synthesis in preadipocytes in a concentration-dependent manner (P <0.05); 3’-hydroxy genistein was similar to rosiglitazone (ROZ) In the 3 and 10 μmol / L groups, a larger amount of lipid droplets appeared in the cytoplasm of preadipocytes (P <0.05 or P <0.01). The mRNA and protein expressions of PPARγ and C / EBPα The protein expression increased (P <0.01). Conclusion 3’-Hydroxy genistein can promote the proliferation and differentiation of preadipocytes and increase the accumulation of lipids during adipocyte differentiation. The mechanism may be related to the promotion of mRNA and protein expression of PPARγ and C / EBPα.