木犀草苷对3T3-L1前脂肪细胞分化和脂代谢的影响

摘要/Abstract

摘要： 目的：研究木犀草苷对3T3-L1前脂肪细胞分化和脂代谢的影响及其作用机制。方法：采用 3T3-L1 前脂肪细胞，利用四甲基偶氮唑蓝（Methyl thiazolyl tetrazolium，MTT）比色分析法检测不同浓度木犀草苷对3T3-L1前脂肪细胞毒性的影响；使用鸡尾酒诱导法诱导细胞分化，然后油红O染色观察脂滴形成情况；测定分化后细胞中甘油三酯、总胆固醇的含量以及相关炎症因子如肿瘤坏死因子（Tumor necrosis factor-α，TNF-α）、白细胞介素6（Interleukin-6，IL-6）、白细胞介素10（Interleukin-10，IL-10）、瘦素(Leptin，LEP)和脂联素(Adiponectin，ADPN)的分泌量；利用Western Blot 技术检测细胞PPARγ、C/EBP-α、SREBP1、PPARα、UCP-1、CPT-1的蛋白表达水平。结果：3T3-L1前脂肪细胞存活率随木犀草苷浓度的升高而降低，木犀草苷浓度在0-60 μg/mL时细胞活力均在80%以上，可进行后续试验。木犀草苷处理组可显著抑制3T3-L1前脂肪细胞的分化，减少脂质积累含量；木犀草苷可下调成脂分化后细胞中TC、TG含量以及炎症因子TNF-α、IL-6和LEP的分泌量，上调IL-10和ADPN的分泌量；木犀草苷能下调PPARγ、C/EBP-α和SREBP1蛋白表达水平，上调PPARα、UCP-1和CPT-1蛋白表达水平。结论：木犀草苷能抑制3T3-L1前脂肪细胞分化及脂代谢，其机制可能是木犀草苷一方面下调PPARγ、C/EBP-α和SREBP1蛋白表达，抑制脂肪合成；另一方面激活PPARα上调下游蛋白促进脂肪消耗；木犀草苷还可能通过调节细胞因子的分泌量，促进细胞脂解，从而改善脂代谢失衡。

关键词: 木犀草苷, 3T3-L1前脂肪细胞, 分化, 脂代谢, 肥胖

Abstract: Objective: Study on the effects and mechanisms of Luteoloside on the differentiation and lipid metabolism of 3T3-L1 preadipocytes. Methods: Using 3T3-L1 preadipocytes, the effect of different concentrations of luteolin on the toxicity of 3T3-L1 preadipocytes was detected using Methyl thiazolyl tetrazolium (MTT) colorimetric analysis; Inducing cell differentiation using cocktail induction method, and then observing lipid droplet formation with Oil Red O staining; The content of triglyceride and total cholesterol in differentiated cells, as well as related inflammatory factors such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), leptin (LEP) and Adiponectin (ADPN) were measured. PPARγ, C/EBP, SREBP1, PPARα, UCP-1, CYP7A1, CPT-1 were analyzed by Western blotting. Conclusion: The survival rate of 3T3-L1 preadipocytes decreases with the increase of Luteoloside concentration, which ranges from 0 to 60 μg/mL When the cell viability is above 80% , subsequent experiments can be conducted. The treatment group with Luteoloside can significantly inhibit the differentiation of 3T3-L1 preadipocytes and reduce lipid accumulation content. Luteoloside can down-regulate the contents of TC and TG, the secretion of inflammatory cytogens TNF-α, IL-6 and LEP in adipogenic and differentiated cells, and up-regulate the secretion of IL-10 and ADPN. Luteoloside down-regulated the protein expression levels of PPARγ, C/EBP and SREBP1, and up-regulated the protein expression levels of PPARα, UCP-1, CYP7A1 and CPT-1. Conclusion: Luteoloside can suppression the differentiation and lipid metabolism of 3T3-L1 preadipocytes, the mechanism may be that Luteoloside down-regulates the protein expression of PPARγ, C/EBP, and SREBP1, and inhibits fat synthesis. On the other hand, it activates PPARα to upregulate downstream proteins and promotes lipolysis. Luteoloside may also inhibit the uptake and accumulation of lipids by cells through inflammatory factors, promote cellular lipid efflux and thus improve lipid metabolism imbalance.

Key words: Luteoloside, 3T3-L1 preadipocytes, differentiation, lipid metabolism, obesity

中图分类号:

TS202.1

杨梦任子怡米思冯丹琦程鑫颖冯雪刘卫华王向红. 木犀草苷对3T3-L1前脂肪细胞分化和脂代谢的影响[J]. 食品科学.

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