球松素查尔酮对C3H10T1/2细胞成脂分化的影响  

Effect of pinostrobin chalcone on adipogenic differentiation of mesenchymal stem cell C3H10T1/2

作　　者：黄伟鹏[1] 黄燕芬[1] 陈建真[1] 金波[1] 谭佳宁 丁志山[1] 

机构地区：[1]浙江中医药大学,浙江杭州310053

出　　处：《中国中药杂志》2017年第12期2339-2344,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81303258);浙江省科技计划项目(2017C33107)

摘　　要：查尔酮类化合物是一种广泛存在于多种药用植物中的黄酮类化合物,具有抑制细胞成脂分化的作用。为研究山核桃叶中提取分离得到的球松素查尔酮对小鼠胚胎间充质干细胞(C3H10T1/2)成脂分化的影响,该实验利用MTS[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,内盐]法检测细胞增殖情况;油红O染色和异丙醇萃取法检测细胞成脂分化情况;GAP-PAP酶法检测细胞中甘油三酯含量;分别采用荧光RT-PCR和Western blot检测成脂分化关键转录因子过氧化物酶体增殖物激活受体γ(PPARγ),CCAAT增强子结合蛋白α(C/EBPα)和分化标志物脂肪型脂肪酸结合蛋白(FABP4)mRNA和蛋白表达。结果显示当球松素查尔酮浓度小于等于50μmol·L^(-1),对C3H10T1/2细胞的增殖活性无影响;油红O染色和异丙醇萃取法及GAP-PAP酶法检测结果表明球松素查尔酮能明显减少C3H10T1/2细胞成脂分化和甘油三酯含量;荧光RT-PCR和Western blot检测表明球松素查尔酮能下调FABP4,PPARγ和C/EBPαmRNA和蛋白的表达(P<0.05或P<0.01)。结果提示,球松素查尔酮能够明显抑制C3H10T1/2细胞成脂分化。Chalcones is a flavonoid wildly presented in many herbs. It has the effect to inhibit cells adipogenie differentiation. In order to study the effect of pinostrobin chalcone extracted and isolated from leaves of hiekoryes on the adipogenic differentiation of mu- rine embryonic mesenehymal stem cell （ C3H10T1/2）, MTS [3-（4,5-dimethyhhiazol-2-yl） -5-（3-carboxymethoxyphenyl） -2-（4-sulfo- phenyl）- 2H-tetrazolium] method was used to detect the cell proliferation; adipogenic differentiation was characterized by oil red O staining and isopropanol extraction; the triglyceride content was detected by GAP-PAP enzyme method; and the C3H10T1/2 cell differ- entiation into adipocytes was also examined by the mRNA and protein expression of PPARγ, C/EBPα and FABP4 by RT-PCR and Western blot respectively. Results indicated that pinostrobin chaleone almost had no effect on cell proliferation activity when the con- centration was less than or equal to 50μmol L-1 ; the oil red O staining, isopropanol extraction and GAP-PAP enzyme method showed that pinostrobin chaleone significantly decreased the C3H10T1/2 adipogenic differentiation and triglyeeride content in the cytoplasm of adipocytes ; the RT-PCR and Western blot analysis showed that pinostrobin chalcone can down-regulate the mRNA and protein levels of FABP4, PPARγ and C/EBPα in C3HIOT1/2 cells（P 〈0. 05 or P 〈 0. 01 ）. The experiment results suggest that pinostrobin chalcone can inhibit C3H10TI/2 adipogenic differentiation.

关 键 词：球松素查尔酮 成脂分化 

分 类 号：R285[医药卫生—中药学]