Rutin Promotes Browning of 3T3-L1 Preadipocytes and Its Mechanism

LYU Yinglan; CHENG Long; SHI Lu; DAI Hongyu; DUAN Yuhui; AN Yongcheng; LI Huimin; WANG Chen; HE Changhao; HUANG Yan; ZHANG Huilin; FU Wanxin; MENG Yanyan; ZHAO Baosheng

doi:10.13422/j.cnki.syfjx.202201727

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Rutin Promotes Browning of 3T3-L1 Preadipocytes and Its Mechanism

更新时间：2023-02-07

- Rutin Promotes Browning of 3T3-L1 Preadipocytes and Its Mechanism
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 5, Pages: 137-143(2023)
- 作者机构：
  
  1.北京中医药大学中药学院，北京 102488
  2.北京中医药大学生命科学学院，北京 102488
  3.北京中医药大学北京中医药研究院，北京 100029
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.202201727
  CLC： R22;R242;R2-031;R285.5
- Published：05 March 2023，
  
  Published Online：20 July 2022，
  
  Received：24 April 2022，
- 稿件说明：
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吕迎兰,程龙,石璐等.芦丁促进3T3-L1前脂肪细胞棕色化及其机制[J].中国实验方剂学杂志,2023,29(05):137-143.

LYU Yinglan,CHENG Long,SHI Lu,et al.Rutin Promotes Browning of 3T3-L1 Preadipocytes and Its Mechanism[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(05):137-143.
吕迎兰,程龙,石璐等.芦丁促进3T3-L1前脂肪细胞棕色化及其机制[J].中国实验方剂学杂志,2023,29(05):137-143. DOI： 10.13422/j.cnki.syfjx.202201727.

LYU Yinglan,CHENG Long,SHI Lu,et al.Rutin Promotes Browning of 3T3-L1 Preadipocytes and Its Mechanism[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(05):137-143. DOI： 10.13422/j.cnki.syfjx.202201727.

摘要

目的

观察芦丁对3T3-L1前脂肪细胞棕色化效应的影响，并探讨其机制。

方法

细胞增殖与活性检测-8（CCK-8）法检测不同浓度芦丁（3.125、6.25、12.5、25、50、100、200 μmol·L

-1

）对3T3-L1细胞活性影响，蛋白免疫印迹法（Western blot）检测不同浓度芦丁（12.5、25、50 μmol·L

-1

）对脂肪细胞产热相关蛋白解偶联蛋白1（UCP1）、PR结构域蛋白16（PRDM16）、过氧化物酶体增殖物激活受体

辅助激活因子-1

（PGC-1

）表达的影响。确定芦丁最佳浓度后，油红O染色观察芦丁对脂肪细胞中脂滴生成的影响，Western blot检测线粒体生物合成标志性蛋白核呼吸因子1（NRF1）、核呼吸因子2（NRF2）和线粒体转录因子A（TFAM）的表达。

结果

与空白组比较，200 μmol·L

-1

芦丁显著抑制3T3-L1细胞活性（

0.01）；在12.5、25、50 μmol·L

-1

浓度下，50 μmol·L

-1

芦丁显著促进产热蛋白（UCP1、PRDM16、PGC-1

）表达（

0.01），确定为最佳浓度。与空白组比较，芦丁50 μmol·L

-1

显著增加3T3-L1细胞线粒体UCP1蛋白的免疫荧光强度（

0.01）及线粒体生物合成标志性蛋白NRF1、NRF2和TFAM的表达（

0.01），显著抑制3T3-L1脂肪细胞脂滴生成（

0.01）。

结论

芦丁可抑制3T3-L1脂肪细胞脂滴沉积，增加产热相关蛋白（UCP1、PRDM16和PGC-1

）及线粒体生物合成标志性蛋白（NRF1、NRF2和TFAM）的表达，从而诱导3T3-L1脂肪细胞棕色化，为开发安全调节白色细胞棕色化的药物提供实验室基础。

Abstract

Objective

To investigate the effect of rutin on the browning of 3T3-L1 preadipocytes and the mechanism.

Method

Cell counting kit-8 （CCK-8） assay was used to detect the effect of different concentration of rutin （3.125， 6.25， 12.5， 25， 50， 100， 200 μmol·L

-1

） on 3T3-L1 cell activity， and Western blot to examine the effect of rutin （12.5， 25， 50 μmol·L

-1

） on the expression of thermogenesis-associated proteins uncoupling protein 1 （UCP1）， PR domain containing 16 （PRDM16） and peroxisome proliferator-activated receptor

coactivator-1

（PGC-1

） in adipocytes. After the optimal concentration of rutin was determined， the effect of rutin on lipid droplet formation in adipocytes was observed based on oil red O staining， and the expression of nuclear respiratory factor 1 （NRF1）， nuclear respiratory factor 2 （NRF2） and mitochondrial transcription factor A （TFAM）， which were the landmark proteins of mitochondrial biosynthesis， was detected by Western blot.

Result

Compared with the blank group， 200 μmol·L

-1

rutin inhibited 3T3-L1 cell activity （

0.01）. Compared with the blank group， at the concentration of 12.5， 25， 50 μmol·L

-1

rutin significantly promoted the expression of thermogenesis-associated proteins （UCP1， PRDM16， and PGC-1

）（

0.01）， which was determined as the optimal concentration. Compared with the blank group， 50 μmol·L

-1

rutin significantly increased the immunofluorescence intensity of mitochondrial UCP1 protein in 3T3-L1 cells （

0.01） and the expression of the markers of mitochondrial biosynthesis （NRF1， NRF2， and TFAM）（

0.01）. In addition， 50 μmol·L

-1

rutin significantly inhibited lipid droplet formation of 3T3-L1 adipocytes （

0.01）.

Conclusion

Rutin inhibited lipid droplet deposition in 3T3-L1 adipocytes and increased the expression of thermogenesis-related proteins （UCP1， PRDM16， and PGC-1

） and markers of mitochondrial biosynthesis （NRF1， NRF2， and TFAM）， thereby inducing the browning of 3T3-L1 adipocytes. This lays a basis for the development of drugs that safely regulate the browning of white cells.

关键词

芦丁3T3-L1前脂肪细胞脂肪细胞棕色化肥胖线粒体生物合成

Keywords

rutin3T3-L1 preadipocytesadipocyte browningobesitymitochondrial biogenesis

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