Gao Min, Ding Zhi-Shan, Shui Yan-Mei, Zhou Fang-Mei, Gao Jian-Li
Zhejiang Chinese Medical University, Hangzhou 310053, China.
Zhongguo Zhong Yao Za Zhi. 2016 Jan;41(1):106-111. doi: 10.4268/cjcmm20160121.
Chrysin is an active flavonoid wildly presented in many herbs. It has the effect to reduce serum lipid. To investigate the effect of chrysin on the adipogenic differentiation of mouse embryonic fibroblasts, methyl thiazolyl tetrazolium (MTT) and crystal violet were used to detect the cytotoxic effect of chrysin on Immortalized mouse embryonic fibroblasts (iMEFs). Propidium iodide (PI) staining combined with flow cytometry (FCM) was employed to detect the effects of different concentrations of chrysin on iMEFs cell cycle. The effect of chrysin on adipogenic differentiation ability of iMEFs was determined by oil red O staining. Semi-quantitative PCR was employed to detect the effect of chrysin on mRNA transcriptional levels of adipogenic differentiation markers, including perilipin 2, adiponectin (adipoq), Fabp4, LPL, MCP-1 and adipogenic differentiation key transcription factor peroxisome proliferators-actiated receptor-gamma 2(PPAR-γ2). Results indicated that chrysin had certain cytotoxic effect for iMEFs in a dose-dependent manner, and the IC₅₀ was identified nearly to 30 μmol•L⁻¹. FCM analysis showed that chrysin could affect the cell-cycle distribution of iMEFs, increasing the ratio of cells in G1 phase. Adipogenic differentiation inducing experiment showed that 30 μmol•L⁻¹ chrysin significantly reduced lipid drops accumulation induced by insulin and dexamethasone. In addition, the mRNA transcriptional levels of PPAR-γ2 and LPL were significantly decreased and mRNA levels of fabp 4, MCP-1, adipoq were also affected after chrysin treatment. The experiment results suggest that chrysin attenuates the adipogenic differentiation capacity of mesenchymal stem cells.
白杨素是一种广泛存在于多种草药中的活性黄酮类化合物。它具有降低血脂的作用。为了研究白杨素对小鼠胚胎成纤维细胞脂肪生成分化的影响，采用甲基噻唑基四氮唑（MTT）和结晶紫检测白杨素对永生化小鼠胚胎成纤维细胞（iMEFs）的细胞毒性作用。采用碘化丙啶（PI）染色结合流式细胞术（FCM）检测不同浓度白杨素对iMEFs细胞周期的影响。通过油红O染色确定白杨素对iMEFs脂肪生成分化能力的影响。采用半定量PCR检测白杨素对脂肪生成分化标志物mRNA转录水平的影响，这些标志物包括脂滴包被蛋白2、脂联素（adipoq）、脂肪酸结合蛋白4（Fabp4）、脂蛋白脂肪酶（LPL）、单核细胞趋化蛋白-1（MCP-1）以及脂肪生成分化关键转录因子过氧化物酶体增殖物激活受体γ2（PPAR-γ2）。结果表明，白杨素对iMEFs具有一定的细胞毒性作用，呈剂量依赖性，其半数抑制浓度（IC₅₀）约为30 μmol•L⁻¹。FCM分析表明，白杨素可影响iMEFs的细胞周期分布，增加G1期细胞比例。脂肪生成分化诱导实验表明，30 μmol•L⁻¹的白杨素可显著减少胰岛素和地塞米松诱导的脂滴积累。此外，白杨素处理后，PPAR-γ2和LPL的mRNA转录水平显著降低，fabp 4、MCP-1、adipoq的mRNA水平也受到影响。实验结果表明，白杨素可减弱间充质干细胞的脂肪生成分化能力。
