白藜芦醇通过ESR1-PI3K信号通路抑制3T3-L1细胞脂滴形成OA北大核心CSTPCD
Resveratrol inhibits lipid droplet formation in 3T3-L1 cells via ESR1-PI3K signaling pathway
[目的]探究白藜芦醇(RES)与雌激素受体1(ESR1)对3T3-L1细胞脂滴形成的影响及调控机制,为RES抑制脂肪沉积潜在机制的研究提供理论依据.[方法]以3T3-L1细胞为试验材料,设计并合成针对ESR1基因的小干扰RNA(siRNA),在3T3-L1细胞培养基中添加RES,以添加等量二甲基亚砜(DMSO)为对照,利用油红O染色、实时荧光定量PCR检测、Western blotting检测等分析RES与ESR1对3T3-L1细胞分化的影响.[结果]与对照组相比,RES能显著(P<0.05)减少3T3-L1细胞脂滴的生成,极显著(P<0.001)上调脂肪分解关键基因ATGL相对表达量,极显著下调(P<0.001)脂肪合成关键基因CEBPα、PPARγ及FAS相对表达量.RES能极显著(P<0.001)增加ESR1基因相对表达量,极显著(P<0.01)增加ESR1蛋白相对表达水平;同时,RES能极显著(P<0.001)下调PI3K信号通路中PI3K和AKT基因相对表达量,极显著(P<0.001)上调FOXO1基因相对表达量.干扰ESR1基因可极显著(P<0.01)增加3T3-L1细胞脂滴的生成,极显著(P<0.001)下调ATGL基因相对表达量,极显著(P<0.001)上调CEBPα、PPARγ及FAS基因相对表达量;加入RES后,干扰ESR1基因的3T3-L1细胞脂滴与未加入RES相比无显著差异(P>0.05,下同),ATGL、CEBPα、PPARγ及FAS基因相对表达量也均无显著差异.干扰ESR1基因能极显著(P<0.001)上调PI3K信号通路中PI3K和AKT基因相对表达量,极显著(P<0.001)下调FOXO1基因相对表达量;加入RES后,PI3K、AKT及FOXO1基因相对表达量与未加入RES相比均无显著差异.[结论]RES能通过ESR1-PI3K信号通路,上调脂肪分解关键基因ATGL相对表达量,下调脂肪合成关键基因CEBPα、PPARγ及FAS相对表达量,从而抑制3T3-L1细胞脂滴形成.
[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.
王倩;杨豹;李银;蒋钦杨;黄艳娜
广西大学动物科学技术学院,广西南宁 530004广西大学动物科学技术学院,广西南宁 530004||广西畜禽繁育与疾病防控重点实验室,广西南宁 530004
畜牧业
白藜芦醇雌激素受体1(ESR1)PI3K信号通路脂滴形成
resveratrolestrogen receptor 1(ESR1)PI3K signaling pathwaylipid droplet formation
《南方农业学报》 2024 (007)
1916-1924 / 9
国家自然科学基金项目(32360839,31760672);广西自然科学基金项目(2022XNSFAA035525) National Natural Science Foundation of China(32360839,31760672);Guangxi Natural Science Foundation(2022XNSFAA035525)
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