Resveratrol inhibits lipid droplet formation in 3T3-L1 cells via ESR1-PI3K signaling pathway
[Objective]To investigate the effects of resveratrol(RES)and estrogen receptor 1(ESR1)on the forma-tion of lipid droplets in 3T3-L1 cells and their regulatory mechanisms,and to provide theoretical basis for study of the po-tential mechanism of RES to inhibit fat deposition.[Method]Using 3T3-L1 cells as test materials,small interfering RNA(siRNA)targeting ESR1 gene was designed and synthesized.RES was added to 3T3-L1 cell culture medium,with the ad-dition of equal amounts of dimethyl sulfoxide(DMSO)as control,the effects of RES and ESR1 on the differentiation of 3T3-L1 cells were analyzed using oil red O staining,real-time fluorescence quantitative PCR assay and Western blotting assay.[Result]Compared with the control group,RES significantly(P<0.05)reduced the production of lipid droplets in 3T3-L1 cells,extremely significantly(P<0.001)up-regulated the relative expression of ATGL,a key gene for lipolysis,and extremely significantly(P<0.001)down-regulated the relative expression of CEBPα,PPARγ and FAS,key genes for adipogenesis.RES extremely significantly(P<0.001)increased the relative expression of ESR1 gene expression,and ex-tremely significantly(P<0.01)increased the relative expression level of ESR1 protein;at the same time,RES extremely significantly(P<0.001)down-regulated the relative expression of PI3K and AKT genes in the PI3K signaling pathway,and extremely significantly(P<0.001)up-regulated the relative expression of FOXO1 gene.Interference with ESR1 gene extremely significantly(P<0.01)increased the generation of lipid droplets in 3T3-L1 cells,extremely significantly(P<0.001)down-regulated the relative expression of ATGL gene,and extremely significantly(P<0.001)up-regulated the relative expression of CEBPα,PPARγ and FAS genes;after the addition of RES,the lipid droplets of 3T3-L1 cells inter-fering with ESR1 gene were not significantly different(P>0.05,the same below)compared with those without RES.There was no significant difference in the relative expression of ATGL,CEBPα,PPARγ and FAS genes.Interfering with ESR1 gene could extremely significantly(P<0.001)up-regulate the relative expression of PI3K and AKT genes in the PI3K signaling pathway,and extremely significantly(P<0.001)down-regulate the relative expression of FOXO1 gene;the relative expression of PI3K,AKT and FOXO1 genes were not significantly different when RES was added compared with that of not adding RES.[Conclusion]Through the ESR1-PI3K signaling pathway,RES can up-regulate the relative expression of ATGL,the key gene of lipolysis,and down-regulate the relative expression of CEBPα,PPARγ and FAS,the key genes of fat synthesis,thereby inhibiting the formation of lipid droplets in 3T3-L1 cells.
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