Inhibitory effect of chloroform extracts from Citrus aurantium L. var. amara Engl. on fat accumulation

Xiao-Yi Li; Yun-Fang Hao; Zhan-Xi Hao; Jian-Guo Jiang; Qiang Liu; Qun Shen; Li Liu; Yan-Kui Yi; Chun-Yan Shen

doi:10.1016/j.phymed.2021.153634

Inhibitory effect of chloroform extracts from Citrus aurantium L. var. amara Engl. on fat accumulation

Phytomedicine. 2021 Sep:90:153634. doi: 10.1016/j.phymed.2021.153634. Epub 2021 Jun 19.

Authors

Xiao-Yi Li¹, Yun-Fang Hao², Zhan-Xi Hao², Jian-Guo Jiang³, Qiang Liu¹, Qun Shen¹, Li Liu¹, Yan-Kui Yi¹, Chun-Yan Shen⁴

Affiliations

¹ School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China.
² College of Food and Bioengineering, South China University of Technology, Guangzhou 510640, China.
³ College of Food and Bioengineering, South China University of Technology, Guangzhou 510640, China. Electronic address: jgjiang@scut.edu.cn.
⁴ School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China; College of Food and Bioengineering, South China University of Technology, Guangzhou 510640, China. Electronic address: shenchunyan@smu.edu.cn.

PMID: 34225246
DOI: 10.1016/j.phymed.2021.153634

Abstract

Background: Excess lipid accumulation can accelerate the development of various metabolic diseases. Blossoms of Citrus aurantium L. var. amara Engl. (CAVA) have been reported to possess inhibitory capacities on lipid deposition. However, the constituents responsible for the observed bioactivity and the underlying mechanisms are still not clearly understood.

Purpose: To screen constituents from blossoms of CAVA with inhibitory effects on lipid accumulation and to explore the action mechanism.

Methods: The chloroform (CHL) extracts are prepared from blossoms of CAVA by fractional extraction and are characterized using LC-MS assay. 3T3-L1 preadipocytes are induced with differentiation medium (DMI) and treated with CHL extracts. High fat diet (HFD)-induced obese mice are further established and administrated with CHL extracts for 12 weeks. Hematoxylin and eosin (HE) staining, Oil Red O staining, ELISA, RT-qPCR, western blot and 16S rRNA gene sequence methods are employed.

Results: 14 compounds are identified in CHL extracts and trigonelline hydrochloride, nobiletin and 7-demethylsuberosin are most abundant. CHL extracts treatment significantly inhibit differentiation of 3T3-L1 cells by regulating expression of preadipocyte factor-1 (Pref-1), fatty acid synthase (FAS) and CCAAT/enhancer binding protein α (C/EBPα). CHL extracts intervention also significantly attenuate features of obesity and improved plasma biochemical profiles in HFD-fed mice. HFD-triggered hepatic steatosis and epididymal adipose tissues (EATs) hypertrophy are also reversed by CHL extracts administration through enhancing antioxidant responses and modulating lipogenesis and energy expenditure-related genes and proteins. 16S rRNA gene sequence data further show that CHL extracts enhance the diversity of gut microbiota. CHL extracts at lower concentrations reduce the ratio of Firmicutes to Bacteroidetes and the abundance of Erysipelotrichaceae. CHL extracts at higher doses markedly increase the abundance of Lachnospiraceae.

Conclusion: These findings suggest that CHL extracts probably suppress lipid accumulation through inhibiting differentiation of 3T3-L1 cells and attenuating metabolic syndromes in HFD-fed mice.

Keywords: Citrus aurantium L. var. amara Engl; Differentiation; Lipid accumulation; Obesity.

MeSH terms

3T3-L1 Cells
Adipogenesis / drug effects*
Animals
Chloroform
Citrus* / chemistry
Diet, High-Fat
Gastrointestinal Microbiome
Lipid Metabolism / drug effects*
Mice
Mice, Inbred C57BL
Mice, Obese
Plant Extracts* / pharmacology
RNA, Ribosomal, 16S

Substances

Plant Extracts
RNA, Ribosomal, 16S
Chloroform