Effects of Triterpenoids from Cyclocarya paliurus(Batal.)Iljinskaja on Adipogenic Differentiation and Regulation of TLR4/MyD88/NF-κB Signaling Pathway in 3T3-L1 Cells
Objective:To observe the effect of triterpenoids from Cyclocarya paliurus(Batal.)Iljinskaja on adipogenic differentiation and regulation of TLR4/MyD88/NF-κB signaling pathway in 3T3-L1 cells.Methods:MTT method was used to determine the effect of triterpenoids from C.paliurus on the proliferation activity of 3T3-L1 cells.Oil-red O staining was used to analyze the fat accumulation of 3T3-L1 cells.Biochemical methods were used to detect the contents of total cholesterol(TC)and triacylglycerol(TG)in 3T3-L1 cells,and ELISA method was used to detect the tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6,IL-4,and IL-10 levels in the supernatant.Real-time PCR was used to determine the mRNA expression levels of Tlr4,Myd88,Nfκbp65,Tnf-α,Il1β,Il6,Il4,and Il10 in 3T3-L1 cells.Western blot was employed to detect the protein expression levels of TLR4,MyD88,IKBα,p-IKBα,IKKβ,p-IKKβ in 3T3-L1 cells and NF-KBp65 in the cytoplasm and nu-cleus of 3T3-L1 cells.Results:Compared with the normal control group,the model control group manifested that the lipid accumulations,levels of TC and TG in cells,and TNF-α,IL-1β,IL-6 contents in in cell supernatants were dramatically elevated;The levels of IL-4 and IL-10 in the supernatants were significantly reduced;the Tlr4,Myd88,Nfκbp65,Tnf-α,Il1β,and Il6 mRNA and TLR4,MyD88,p-IκBα,p-IKKβ,NF-κBp65 in 3T3-L1 cells and nucleus,p-IKBα/IKBα and p-IKKβ/IKKβ ratios were markedly hoisted;the mRNA expression of Il4,Il10 and protein expression of NF-KBp65 in cytoplasm were prominently lessened(P＜0.01);Compared with the model control group,the different dosage groups of the triterpenoids from C.paliurus significantly inhibited the lipid accumulation in 3T3-L1 cells,decreased the con-tent of TC and TG in cells and the level of TNF-α,IL-1β,and IL-6 in the supernatant,elevated the IL-4 and IL-10 contents in the superna-tant,reduced the mRNA expression of Tlr4,Myd88,Nfκbp65,Tnf-α,Il1β,and Il6,the protein expression of TLR4,MyD88,NF-KBp65 in nu-cleus,and p-IκBα/IKBα and p-IKKβ/IKKβ ratios in 3T3-L1 cells,and increased the mRNA expression of Il4,Il10 and protein expression of NF-KBp65 in cytoplasm(P＜0.05 or P＜0.01).Conclusion:The triterpenoids from C.paliurus have a good inhibitory effect on adipogenic differentiation of 3T3-L1 cells.Its mechanism is related to inhibition of TLR4/MyD88/NF-κB signaling pathway,reduction of adipocyte in-flammatory response,inhibition of TNF-α,IL-1β,IL-6,TC,and TG productions and lipid accumulation,and acceleration of the IL-4 and IL-10 productions.The above results demonstrate the good application prospects of triterpenoids from C.paliurus in the prevention and treatment of obesity.
Triterpenoids from C.paliurus3T3-L1 cellsLipid accumulationsignaling pathway
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