黄伟鹏+黄燕芬+陈建真+金波+谭佳宁+丁志山

[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50 μmol·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/EBPα mRNA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。

[关键词] 球松素查尔酮； 成脂分化

[Abstract] Chalcones is a flavonoid wildly presented in many herbs. It has the effect to inhibit cells adipogenic differentiation. In order to study the effect of pinostrobin chalcone extracted and isolated from leaves of hickoryes on the adipogenic differentiation of murine embryonic mesenchymal stem cell （C3H10T1/2）， MTS [3-（4，5-dimethylthiazol-2-yl）-5-（3-carboxymethoxyphenyl）-2-（4-sulfophenyl）- 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 differentiation 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 chalcone almost had no effect on cell proliferation activity when the concentration was less than or equal to 50 μmol·L^-1； the oil red O staining， isopropanol extraction and GAP-PAP enzyme method showed that pinostrobin chalcone significantly decreased the C3H10T1/2 adipogenic differentiation and triglyceride 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 C3H10T1/2 cells（P<0.05 or P<0.01）. The experiment results suggest that pinostrobin chalcone can inhibit C3H10T1/2 adipogenic differentiation.

[Key words] pinostrobin chalcone； adipogenic differentiation

随着生活水平提高，肥胖已经成为影响人类健康的重要因素。肥胖症是由于脂肪细胞过度分化和脂肪颗粒在脂肪细胞中过度沉积而引起的慢性炎症性疾病[1]。肥胖被視为心脑血管疾病、2型糖尿病、消化系统疾病等的危险因素[2]。间充质干细胞是一类可进行自我复制并能向脂肪细胞、成骨细胞、软骨细胞等多方向分化的成体干细胞[3]。间充质干细胞受PPARγ和CCAAT调节/增强子结合蛋白（C/EBP）等转录因子的影响，导致脂结合蛋白4、脂联素等成脂分化标志基因表达增加，进而使胞内甘油三酯逐步累积，形成脂肪细胞[4]。……