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目的探讨姜黄素对猪脂肪间充质干细胞(AMSCs)成脂分化的影响,及姜黄素调控成脂分化的机制。方法用不同浓度姜黄素处理猪AMSCs,用MTT比色法检测姜黄素对猪AMSCs增殖的影响,用油红O染色提取法检测对成脂分化的程度,用实时定量聚合酶链反应(Real-time PCR)检测对Krüppel样因子(KLF2)和过氧化物酶增殖物激活受体γ(PPARγ)2 mRNA表达的影响。结果当姜黄素浓度为1、5、10μmol/L用于猪AMSCs时对,AMSCs增殖无明显影响,但高于15μmol/L时,则具有显著的抑制作用(P<0.05);当姜黄素用于猪AMSCs的成脂诱导分化时,1~30μmol/L的浓度即能显著抑制成脂分化(P<0.05),其中25μmol/L浓度获得68.19%的高抑制率;当25μmol/L姜黄素用于检测KLF2和PPARγ2 mRNA的表达,诱导分化2、8和16d的KLF2 mRNA的表达上调率分别达到48.37%、20.56%和9.64%,而PPARγ2 mRNA的表达下调率分别达到16.01%、35.93%和27.27%。结论姜黄素具有抑制猪AMSCs增殖和成脂分化作用,该抑制作用与姜黄素促进KLF2 mRNA的表达和抑制PPARγ2 mRNA的表达相关。
Objective To investigate the effect of curcumin on adipogenic differentiation of porcine adipose-derived mesenchymal stem cells (AMSCs) and the mechanism of curcumin-mediated adipogenic differentiation. Methods The pig AMSCs were treated with different concentrations of curcumin. The effects of curcumin on the proliferation of pig AMSCs were detected by MTT colorimetric assay. The degree of adipogenic differentiation was detected by oil red O staining and the real-time quantitative polymerase chain reaction (Real- Time PCR was used to detect the effect of Krüppel-like factor (KLF2) and peroxisome proliferator-activated receptor γ (PPARγ) 2 mRNA expression. Results When curcumin concentration was 1,5,10 μmol/L for pig AMSCs, there was no significant effect on the proliferation of AMSCs, but when it was higher than 15 μmol/L, it had a significant inhibitory effect (P<0.05); when curcumin was used When adipogenic differentiation of swine AMSCs was induced, the concentration of 1-30 μmol/L could significantly inhibit adipogenic differentiation (P<0.05), and 25 μmol/L concentration had a high inhibition rate of 68.19%. When 25 μmol/L curcumin was used The expression of KLF2 and PPARγ2 mRNA was detected. The up-regulation rates of KLF2 mRNA expression on the 2nd, 8th, and 16th day of differentiation were 48.37%, 20.56%, and 9.64%, respectively, while the down-regulation rates of PPARγ2 mRNA were 16.01%, 35.93%, and 27.27, respectively. %. Conclusion Curcumin can inhibit the proliferation and adipogenic differentiation of porcine AMSCs. The inhibitory effects of curcumin on KLF2 mRNA expression and PPARγ2 mRNA expression are related to curcumin.