芦丁抑制3T3-L1前脂肪细胞分化并促进米色脂肪细胞形成

Rutin inhibits 3T3-L1 preadipocytes differentiation and promotes beige adipogenesis

目的探讨芦丁对米色脂肪细胞形成的影响。方法用不同浓度芦丁处理3T3-L1前脂肪细胞;细胞计数盒(CCK8)检测不同浓度芦丁对细胞活性影响,油红O染色观察脂滴大小;Western blot检测过氧化物酶增殖物激活受体–γ(PPAR-γ)、aP2及解偶联蛋白1(UCP1)表达;RT-qPCR检测UCP1、PRDM16、Dio2表达;细胞免疫荧光检测线粒体变化。结果芦丁在0~250μmol/L范围内对细胞活性无明显影响,且以药物依赖性方式减少脂滴生成;与对照组[aP2(1.12±0.03)]、[PPAR-γ(0.87±0.02)]比较,50、100、250μmol/L芦丁使a P2[分别为(0.87±0.03)、(0.64±0.01)、(0.64±0.05)]、PPAR-γ[分别为(0.73±0.02)、(0.46±0.02)、(0.43±0.05)]蛋白表达降低,使UCP1蛋白表达[分别为(0.58±0.03)、(0.42±0.02)、(0.34±0.01)]增加;与对照组比较,50μmol/L芦丁可使UCP1、PRDM16、Dio2表达[分别为(7.02±0.24)、(2.09±0.06)、(10.40±0.93)]升高;细胞免疫荧光显示,3T3-L1前脂肪细胞内线粒体含量增加。结论芦丁可减少3T3-L1前脂肪细胞脂滴生成,并促进米色脂肪细胞形成。

Objective To investigate the effect of rutin on beige adipogenesis development.Methods Preadipocytes(3T3-L1)were treated with rutin at different concentrations.Cell Counting Kit-8(CCK8)was used to assess the effect of different concentrations of rutin on cell viability.Oil red O staining was used to observe lipid droplet size.The expressions of peroxisome proliferators-activated receptors(PPAR-γ),human adipocyte lipid-binding protein(aP2)and uncoupling protein 1(UCP1)were detected with Western blot.The mRNA expressions of the UCP1,PR domain-containing the 16-like protein(PRDM16)and deiodinase type 2(Dio2)genes were determined using reverse transcription quantitative PCR(RT-qPCR).Changes in mitochondria were detected by cellular immunofluorescence.Results No significant variation in cell viability was observed but lipid droplet formation was reduced in a dose-dependent manner in the 3T3-L1 cells treated with rutin at the dosages 0–250μmol/L.Compared with those in the negative control cells,decreased protein expressions of aP2(0.87±0.03,0.64±0.01,and 0.64±0.05 vs.1.12±0.03)and PPAR-γ(0.73±0.02,0.46±0.02,and 0.43±0.05 vs.0.87±0.02)but increased UCP1 protein(0.58±0.03,0.42±0.02,and 0.34±0.01)were detected in the preadipocytes treated with rutin at dosages of 50,100,and 250μmol/L.Up-regulated expressions of UCP1(7.02±0.24),PRDM16(2.09±0.06),and Dio2(10.40±0.93)genes were detected in the 3T3-L1 cells treated with rutin of 50μmol/L in comparison with that of control cells.Cellular immunofluorescence assay revealed increased number of mitochondrial in 3T3-L1 preadipocytes exposed to 50μmol/L rutin.Conclusion Rutin can reduce the formation of lipid droplets and promote beige adipogenesis in 3T3-L1 preadipocytes.