川续断皂苷Ⅵ对3T3-L1细胞的增殖和分化影响  

Effect of asperosaponin Ⅵ on proliferation and differentiation of 3T3-L1 cells

作　　者：陈小宇[1] 祝爱珍[1] 刘成成[1] 朱锦灿[1] 刘革修[1] 

机构地区：[1]暨南大学血液病研究所,广东广州510632

出　　处：《中国药理学通报》2013年第8期1150-1154,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81270568)

摘　　要：目的研究川续断皂苷Ⅵ(asperosaponinⅥ,ASAⅥ)对3T3-L1前脂肪细胞的增殖及成脂分化影响,以进一步了解其药理作用。方法以3T3-L1前脂肪细胞为靶细胞,通过噻唑蓝(MTT)法观察不同浓度川续断皂苷Ⅵ对3T3-L1细胞增殖能力的影响;不同浓度(0、10、25、50μmol·L-1)川续断皂苷Ⅵ干预3T3-L1细胞的成脂诱导分化,油红O染色检测3T3-L1细胞分化程度;试剂盒测定各组培养液中葡萄糖及游离脂肪酸含量;荧光定量PCR(Q-PCR)检测过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activatedreceptorγ,PPARγ)、CCAAT增强子结合蛋白(CCAAT/enhan-cer-binding proteinα,C/EBPα)mRNA表达水平。结果 MTT法测定表明不同浓度川续断皂苷Ⅵ对3T3-L1细胞增殖能力的影响不同,其中10μmol·L-1浓度组作用24 h后促增殖作用最明显,随着川续断皂苷Ⅵ浓度增加,促增殖效应降低;川续断皂苷Ⅵ能够剂量依赖性的抑制3T3-L1细胞成脂,减少细胞内脂滴聚集,抑制细胞对葡萄糖的吸收利用,减少游离脂肪酸形成,下调成脂过程中PPARγ和C/EBPαmRNA表达。结论川续断皂苷Ⅵ具有促进3T3-L1细胞增殖作用,并通过下调PPARγ和C/EBPα基因抑制3T3-L1细胞成脂分化。Aim To explore the effects of asperosaponin VI on proliferation and differentiation of 3T3-L1 preadipocytes. Methods 3T3-L1 preadipocytes were taken as the target cells, MTT assay was applied to investigate the proliferation of 3T3-LI cells for 72 h. The cells were cultured with inducer and 0, 10, 25, 50 μmol· L-1 ASAVI respectively. On the 6th day of in- duced-differentiation, the differentiation of 3T3-L1 cells was detected by oil red O staining. The concentration of glucose and non-esterified fatty acid in culture supernatant was measured by commercial kits. The expression of PPAR γ and C/EBP α mRNA was determined with real-time fluorescence quantitative PCR（Q- PCR）. Results ASA VI at different concentrations had various effects on proliferation of 3T3-L1 cells, the promoting effects of ASA VI were the best at the 24th hour in 10 μmol· L-1 group. With the concentration increasing, the promoting effects descended. ASA VI inhibited 3T3-L1 preadipocytes differentiation in a dose-dependent manner, and decreased lipid accumulation, inhibited the intaking of glucose and reduced the production of non-esterified fatty acid in culture su- pernatant. At molecular level, the expression of transcription factors, PPARγand C/EBP α, was reduced by ASA VI during adipogenesis. Conclusion These results indicate that ASA VI can effectively promote proliferation and inhibit 3T3-L1 preadipocytes differentiation.

关 键 词：川续断皂苷Ⅵ 3T3-L1前脂肪细胞 成脂分化 细胞增殖 间充质干细胞 过氧化物酶体增殖物激活受体Γ CCAAT增强子结合蛋白α 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]