葛根素对松辽黑猪前体脂肪细胞成脂分化的影响

为探究葛根素对松辽黑猪前体脂肪细胞成脂分化的调控作用,在细胞诱导液中分别添加0、10、20、40、60和80 μmol/L葛根素进行成脂诱导分化,用油红O染色法和甘油三酯酶法检测脂肪细胞分化过程中脂滴聚集情况和甘油三酯含量以考察脂质沉积和分化效果,并确定葛根素的最佳添加浓度;用实时荧光定量PCR检测对照组(0 μmol/L)和最佳葛根素浓度添加组成脂标志基因细胞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)及成脂分化基因乙酰辅酶A羧化酶(ACC)、脂肪酸结合蛋白(FABP4)、应激蛋白(TRIB)和叉头框蛋白O1 (FOXO1)的mRNA的表达水平,用Western blotting检测PPARγ和C/EBPα的蛋白表达水平。结果表明,与对照组相比,20、40和60 μmol/L葛根素均显著增加脂滴和甘油三酯含量(P<0.05),且40 μmol/L葛根素效果最佳;实时荧光定量PCR结果表明,与对照组相比,40 μmol/L葛根素显著上调成脂标志基因PPARγ、C/EBPα及成脂分化基因ACC、FABP4、FOXO1和TRIB的表达(P<0.05);Western blotting结果显示,与对照组相比,40 μmol/L葛根素显著增加PPARγ蛋白表达(P<0.05)。综上所述,40 μmol/L葛根素能够促进松辽黑猪前体脂肪细胞的成脂分化和脂质沉积。

Effect of Puerarin on Adipogenic Differentiation of Preadipocytes in Songliao Black Pig

In order to investigate the regulation effect of puerarin on the adipogenic differentiation of precursor adipocytes in Songliao Black pig, 0, 10, 20, 40, 60 and 80 μmol/L puerarin were added into the cell induction solution to induce adipogenic differentiation, respectively.In order to determine the effect of lipid deposition and differentiation, oil red O staining and triglyceride assay were used to detect the accumulation of lipid droplets and triglyceride content during adipocyte differentiation, and to determine the best concentration of puerarin.The mRNA expression levels of lipid marker genes cell peroxisome proliferator-activated receptor γ(PPARγ), CCAAT-enhancer binding protein α(C/EBPα) and genes related to adipogenic differentiation like acetyl-CoA carboxylase (ACC), fatty acid binding protein(FABP4), stress protein(TRIB) and forked frame protein O1 (FOXO1) were detected by Real-time quantitative PCR in the control group and the optimal puerarin concentration group.The protein expression levels of PPARγ and C/EBPα were detected by Western blotting.The results showed that compared with control group, 20, 40 and 60 μmol/L puerarin significantly increased the content of lipid droplets and triglycerides (P<0.05), and 40 μmol/L puerarin had the best effect.Real-time quantitative PCR results showed that compared with control group, 40 μmol/L puerarin significantly up-regulated the expression of adipogenic marker genes PPARγ and C/EBPα and adipogenic differentiation genes ACC, FABP4, FOXO1 and TRIB (P<0.05).Western blotting results showed that compared with control group, 40 μmol/L puerarin significantly increased the expression of PPARγ protein (P<0.05).In conclusion, 40 μmol/L puerarin could promote the adipogenic differentiation and lipid deposition of preadipocytes in Songliao Black pigs.