Combinational Effect Of Pine Needle Polysaccharide And Kudzu Flavonoids On Cell Differentiation And Lipid Metabolism In 3T3-L1 Cells

Pine needle polysaccharide(PNP) reduced the serum triglyceride levels, body fat and weight gain. Kudzu flavonoids(KF) regulated fat accumulation by regulating the expression of lipid metabolism related genes in adipocytes. At present, the combinational effect of PNP and KF on lipid metabolism had not been reported. This present study is to investigate the effect of PNP and KF combination on cell differentiation and lipid metabolism in 3T3-L1 cells.MTT method was used to detect the effect of PNP and KF on cell proliferation.Intracellular triglyceride(TG) content and the content of adipocytokines(ADP, LEP,TNF-?, IL-6) were determined after cell differentiation at day 8. The TG content in culture medium after norepinephrine(NA)-induced fat decomposition was detected. The expression of lipid metabolism related genes(PPAR?2, C/EBP?, SREBP1 c, ACC?, FAS,HMG-CoAR, ATGL, HSL, ACO, CAT, ADP, LEP, IL-6, Glut4, LKB1, CaMKK, AMPK) and proteins(ACC, FAS, ATGL, HSL, p-HSL, Glut4, LKB1, AMPK, p-AMPK) were detected of different differentiation days in 3T3-L1 cells. Results were as followed:(1) PNP and KF combination markedly inhibited cell proliferation, and was dose dependent relationship with the concentration of KF.(2) PNP and KF combination significantly reduced intracellular TG content, reduced the concentration of LEP, TNF-? and IL-6, with augmented ADP secretion level at day 8 of cell differentiation. PNP and KF have synergistic effect.(3) PNP and KF combination significantly promoted lipid deposition induced by NA,and was positive associated with PNP concentration.(4) PNP and KF combination markedly up-regulated the mRNA expression of LKB1,AMPK, ATGL, HSL, ACO, CAT, ADP and Glut4, meanwhile reduced the expression of PPAR?2, C/EBP?, SREBP1 c, ACC?, FAS, HMG-CoAR, LEP and IL-6, with no obvious effect of CaMKK.(5) PNP and KF combination significantly inhibited the protein expression of FAS and ACC, and up-regulated the protein expression of LKB1, AMPK, p-AMPK, ATGL, HSL and p-HSL in 3T3-L1 cells.PNP and KF combination inhibited intracellular lipid synthesis, promoted fatty acid decomposition, reduced the TG content through regulating the expression of lipid metabolism related genes and proteins as well as the secretion of adipocytokines,suggesting that PNP and KF combination have good slimming effect.