东紫苏精油对间充质干细胞C3H10T1/2成脂分化的影响及机制

Effect and Mechanism of Essential Oil from Elsholtiza bodinieri Vaniot on Adipogenic Differentiation of Mesenchymal Stem Cells C3H10T1/2

为探讨东紫苏精油对小鼠胚胎间充质干细胞C3H10T1/2成脂分化的作用及降脂机制,利用噻唑蓝[the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT]法检测细胞成活率,以界定浓度的适宜范围;用油红O染色法对东紫苏精油作用后的细胞成脂分化作用进行分析;甘油磷酸氧化酶—过氧化物酶(glycerol phosphate oxidase-p-aminophenazone,GPO-PAP)法、胆固醇氧化酶、过氧化物酶和4-氨基安替比林苯酚(cholesterol oxidase,peroxidase and 4-aminoantipyrine phenol,COD-PAP)酶法检测东紫苏精油处理后细胞内甘油三酯、胆固醇含量的变化;荧光定量聚合酶链式反应(polymerase chain reaction,PCR)和蛋白质印迹(western blot)分别检测过氧化物酶体增殖物激活受体γ(peroxide proliferator-activated receptorγ,PPARγ)、CCAAT增强子结合蛋白α(CCAAT/enhancer-binding proteinsα,C/EBPα)、脂肪酸结合蛋白4(fatty acid-binding protein4,FABP4)mRNA和相关蛋白在成脂分化中的表达水平。结果表明:与空白对照组相比,高脂模型组的总胆固醇(total cholesterol,TC)和甘油三酯(triacylglyceride,TG)含量显著增加(P<0.05),小鼠胚胎间充质干细胞C3H10T1/2中PPARγ、C/EBPα、FABP4 mRNA和蛋白表达明显增加。与高脂模型组相比,中剂量(50μg/mL)和高剂量(100μg/mL)精油组TC和TG含量明显下降,小鼠胚胎间充质干细胞C3H10T1/2中PPARγ、C/EBPα、FABP4 mRNA和蛋白表达明显下降。试验结果表明,东紫苏精油可能通过调控PPARγ/C/EBPα/FABP4信号通路发挥对C3H10T1/2细胞成脂分化的抑制作用,达到减脂目的。

This paper investigated the effect of essential oil from Elsholtiza bodinieri vaniot on adipogenic dif⁃ferentiation and lipid⁃reducing mechanism of mouse embryonic mesenchymal stem cells C3H10T1/2.The sur⁃vival of cells was measured by the 3⁃(4,5⁃dimethylthiazol⁃2⁃yl)⁃2,5⁃diphenyltetrazolium bromide(MTT)method to define the appropriate range of concentration.After treatment with E.bodinieri essential oil,the oil red O staining was performed to analyze the cell adipogenic differentiation.3⁃(4,5⁃dimethylthiazol⁃2⁃yl)⁃2,5⁃diphenyltetrazolium bromide(GPO⁃PAP),and cholesterol oxidase,peroxidase and 4⁃aminoantipyrine phenol(COD⁃PAP)were used to detect the changes in triglyceride(TG)and total cholesterol(TC)in cells treated with essential oil.The mRNA and protein expressions of peroxide proliferator⁃activated receptorγ(PPARγ),CCAAT/enhancer⁃binding proteinsα(C/EBPα),and fatty acid⁃binding protein 4(FABP4)in lipid differentia⁃tion were detected by quantitative polymerase chain reaction(PCR)and western blot,respectively.The re⁃sults showed that compared with the normal blank control group,the high⁃fat model group had increased TC and TG(P<0.05),and the mRNA and protein expressions of PPARγ,C/EBPα,and FABP4 in C3H10T1/2 cells were up⁃regulated.Compared with the conditions in high⁃fat model group,the TC and TG in middle⁃dose(50μg/mL)and high⁃dose(100μg/mL)essential oil group were decreased,and the mRNA and protein ex⁃pressions of PPARγ,C/EBPα,and FABP4 in C3H10T1/2 cells were down⁃regulated.The results indicated that the essential oil may inhibit the adipogenic differentiation of C3H10T1/2 cells by regulating PPARγ/C/EBPα/FABP4 signaling pathway to achieve lipid reduction.