川续断皂苷Ⅵ对小鼠骨髓基质细胞ST-2成脂分化的影响及其机制研究  

Effect of Asperosaponin Ⅵ on adipocyte differentiation in ST-2 cells and its underlying mechanisms

作　　者：王海晓[1] 崔壮[2] 王宝利[3] 边育红[1] 郑纺[1] 

机构地区：[1]天津中医药大学中西医结合学院,300193 [2]天津医科大学公共卫生学院 [3]天津医科大学代谢病医院内分泌研究所

出　　处：《天津医药》2015年第12期1345-1348,共4页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(81102543;81200611);中国博士后基金面上项目(2012M520588)

摘　　要：目的观察川续断皂苷Ⅵ(AsperosaponinⅥ,ASAⅥ)对脂肪细胞分化的影响及Wnt通路的调节。方法以小鼠骨髓基质细胞系ST-2为研究对象,将其分为对照组、成脂分化组以及4个不同剂量的ASAⅥ组。其中对照组加入溶媒,成脂分化组加入成脂诱导分化试剂,ASAⅥ组除加入成脂诱导分化试剂外,使用不同浓度(10^(-7)、10^(-6)、10^(-5)、10^(-4)mol/L)的ASAⅥ干预细胞。各组细胞处理5d后,行油红O染色,观察脂滴形成情况并计算成脂率进行定量分析;荧光定量PCR(FQ-PCR)检测成脂相关基因PPARγ、FABP4和Wnt/β-连环素(β-catenin)通路蛋白β-catenin的mRNA表达水平;Wnt通路抑制剂DKK1干预诱导成脂分化的ST-2细胞5 d,FQ-PCR检测ASAⅥ所调节的PPARγ、FABP4和β-catenin mRNA表达水平。结果与成脂分化组相比,10^(-5)mol/L和10^(-4)mol/L ASAⅥ组中分化的脂肪细胞显著减少,10^(-5)、10^(-4)mol/L ASAⅥ明显抑制PPARγ、FABP4的mRNA表达,但上调β-catenin mRNA表达。DKK1能够逆转ASAⅥ对ST-2细胞成脂分化的抑制作用,促进PPARγ、FABP4的mRNA表达,抑制β-catenin的mRNA表达。结论 ASAⅥ能显著抑制ST-2细胞的成脂分化,这一作用可能是通过Wnt/β-catenin通路的激活所介导的。Objective The effect of Asperosaponin Ⅵ（ASAⅥ）on adipocyte differentiation and the involvement of Wnt signal pathway was investigated. Methods The murine bone marrow stromal cell line ST-2 were divided into 6 groups：control group, adipocyte differentiation group, and 4 different doses of ASAⅥgroups. Control group was exposed to the vehicle, adipocyte differentiation group was exposed to adipogenic reagent, and those 4 ASAⅥgroups were treated with different concentration（10-7, 10-6, 10-5, 10-4 mol/L）of ASAⅥafter adipocyte differentiation induction. 5 days later, oil red O staining was performed to calculate adipocyte rate. Then mRNA transcription levels of PPARγ, FABP4 genes andβ-catenin that were Wnt/β-catenin signaling pathway proteins were examined by FQ-PCR. Then Wnt pathway inhibitor DKK1 was supplemented into ST-2 cells treated with 10-4 mol/L ASAⅥfor 5 days. After that FQ-PCR was used to detect whether tran？scription levels of PPARγ, FABP4 andβ-catenin in ST-2 cells were changed. Results Compared with adipocyte differenti？ation group 10-5 mol/L and 10-4 mol/L ASAⅥtreatments greatly down-regulated the number of lipid droplets and markedly inhibited transcription levels of adipocyte characterization transcription factors included PPARγ, FABP4 while up-regulat？ed transcription level ofβ-catenin in ST-2 cells. DKK1 can reverse the inhibitory effect of ASAⅥon adipocyte differentia？tion in ST-2 adipocyte. The transcription levels of PPARγand FABP4 were up-regulated significantly while transcription level ofβ-catenin was inhibited. Conclusion ASAⅥblocks adipocyte differentiation in ST-2 cells which might be medi？ated through activating Wnt/β-catenin signaling pathway.

关 键 词：成脂分化 Β连环素 川续断皂苷Ⅵ 骨髓基质细胞 WNT信号通路 

分 类 号：R285.5[医药卫生—中药学]