高良姜二苯庚烷化合物对3T3-L1前脂肪细胞分化的影响和机制研究

Effect and Mechanism of Galangal Diphenylheptane Compound on 3T3-L1 Preadipocyte Differentiation

目的探讨高良姜二苯庚烷化合物对3T3-L1前脂肪细胞分化的影响和机制。方法本研究选取2022年9月—2023年2月珠海市人民医院·暨南大学附属珠海医院3T3-L1前脂肪细胞为研究对象,使用MTT法评估高良姜二苯庚烷化合物对3T3-L1细胞增殖的影响。通过油红O染色检测分析高良姜二苯庚烷化合物对3T3-L1细胞分化的影响。应用实时定量聚合酶链锁反应(Polymerase Chain Reaction,PCR)方法检测高良姜二苯庚烷化合物对过氧化物酶体增殖物激活受体γ(Peroxisome Proliferator Activated Receptorγ,PPARγ)和CCAAT/增强子结合蛋白α(CCAAT/Enhancer Binding Proteinα,C/EBPα)基因mRNA表达的调控作用。通过Western blot分析检测高良姜二苯庚烷化合物对PPARγ和C/EBPα蛋白表达的影响。结果高良姜二苯庚化合物(0、0.05、0.2、0.4 g/L)在24、48 h对前脂肪细胞的生长表现为增殖趋势,呈现一定的量效关系,其中高良姜二苯庚烷化合物浓度为0.4 g/L时3T3-L1增殖[(7.02±0.35)、(17.23±0.95)g/L]高于与对照组,差异有统计学意义(t=13.794、10.843,P均<0.05)。高良姜二苯庚烷化合物呈剂量依赖性地抑制了3T3-L1细胞的增殖。油红O染色结果显示,高良姜二苯庚烷化合物处理显著抑制了3T3-L1细胞的脂肪细胞分化,并减少了脂滴的积累;100μmol/L处理组,PPARγ和C/EBPα的mRNA表达水平显著降低。高良姜二苯庚烷化合物通过抑制PPARγ和C/EBPα基因的表达,调控了3T3-L1前脂肪细胞的分化过程,从而减少了脂肪滴的积累。结论高良姜二苯庚烷化合物抑制3T3-L1前脂肪细胞分化,其机制与PPARr/EBPα的低表达有关。

Objective To investigate the effect and mechanism of galangal diphenylheptane compound on 3T3-L1 preadipocyte differentiation.Methods This study selected 3T3-L1 preadipocytes from Zhuhai People′s Hospital and Jinan University Affiliated Zhuhai Hospital from September 2022 to February 2023 as the research subjects,and used the MTT method to evaluate the effect of galangal diphenylheptane compounds on the proliferation of 3T3-L1 cells.The effect of Alpinia officinalis diphenylheptane compounds on 3T3-L1 cell differentiation was analyzed through O il Red O staining.The regulatory effect of Galangal diphenylheptane compounds on mRNA expression of peroxisome proliferator activated receptorγ(PPARγ)and CCAAT/enhancer binding proteinα(C/EBPα)were detected through polymerase chain reaction(PCR)method.The effect of Galangal diphenylheptane compounds on the protein expression of PPARγand C/EBPαthrough Western blot analysis.Results Galangal diphenylheptyl compounds(0,0.05,0.2,0.4 g/L)showed a proliferation trend on the growth of preadipocytes at 24 h and 48 h,showing a certain dose-effect relationship.When the concentration was 0.4 g/L,the difference was statistically significant compared with the control group(7.02±0.35)g/L and(17.23±0.95)g/L(t=13.794,10.843,both P<0.05).Galangal diphenylheptane compounds inhibited the proliferation of 3T3-L1 cells in a dose-dependent manner.The results of oil red O staining showed that the diphenylheptane compound of galangal significantly inhibited the adipocyte differentiation of 3T3-L1 cells and reduced the accumulation of lipid droplets.mRNA expression levels of PPARγand C/EBPαwere significantly decreased in the 100μmol/L treatment group.Galangal diphenylheptane compounds regulate the differentiation of 3T3-L1 preadipocytes by inhibiting the expression of PPARγand C/EBPαgenes,thereby reducing the accumulation of fat droplets.Conclusion Galangal diphenylheptane compounds inhibit 3T3-L1 preadipocyte differentiation,and the mechanism is related to the low expression of PPARr/EBPα.