Regulatory effect of Diwu Yanggan Decoction on lysoglycerophospholipids in circulating exosomes in a mouse model of nonalcoholic fatty liver disease

doi:10.12122/j.issn.1673-4254.2024.07.18

Abstract

Abstract:

Objective To evaluate the regulatory effect of Diwu Yanggan (DWYG) Decoction on lysoglycerophospholipids (Lyso-GPLs) in circulating exosomes in a mouse model of nonalcoholic fatty liver disease (NAFLD). Methods Circulating exosomes isolated from mouse serum by size exclusion chromatography were morphologically characterized using transmission electron microscope and examined for surface markers CD9, CD63 and TSG101 using Western blotting. Twenty-four male Kunming mice were randomized into 3 groups for normal feeding (control, n=8) or high-fat diet feeding for 1 week to induce NAFLD, after which the latter mice were given DWYG decoction (treatment group, n=8) or normal saline (model group, n=8) by gavage for 4 weeks. After the last treatment, blood samples were collected from the mice for testing serum TC, HDL-C, LDL-C, ALT and AST levels and isolating circulating exosomes. Using multivariate statistical analysis based on targeted metabolomics strategy, the potential biomarkers for Lyso-GPLs in the exosomes were screened. Results The isolated exosomes about 100 nm in size had a typical saucer-like structure with distinct double-layer membranes and a mean particle size of 137.5 nm and expressed the specific surface marker proteins CD9, CD63 and TSG101. The mouse models of NAFLD had significantly increased serum levels of TC, HDL-C, LDL-C and AST and lowered serum ALT level. A total of 43 Lyso-GPLs with significant reduction after DWYG Decoction treatment were identified in NAFLD mice. Conclusion DWYG Decoction can regulate Lyso-GPLs in circulating exosomes in NAFLD mice, which provides a new clue for studying the therapeutic mechanism of DWYG Decoction for liver disease.

Key words: Diwu Yanggan Decoction, circulating exosomes, UPLC-QTOF-MS/MS, lysoglycerophospholipid, nonalcoholic fatty liver disease

Guangya CHEN, Xingliang XIANG, Zhaoxiang ZENG, Rongzeng HUANG, Shuna JIN, Mingzhong XIAO, Chengwu SONG. Regulatory effect of Diwu Yanggan Decoction on lysoglycerophospholipids in circulating exosomes in a mouse model of nonalcoholic fatty liver disease[J]. Journal of Southern Medical University, 2024, 44(7): 1382-1388.

Figures/Tables 5

Fig.2 Chromatogram of the metabolite profile of 102 lysoglycerophospholipids.

Tab.1 Physiological and biochemical parameters of the mice (Mean±SD, n=8)

Parameter	Normal	Model	DWYG
Physiological parameter
Body weight (g)	48.25±4.38	50.13±4.13	46.90±3.97
Liver weight (g)	2.01±0.38	2.37±0.22	1.94±0.30^#
Liver/body index	4.14±0.48	4.74±0.45*	4.13±0.44^#
Biochemical parameters
AST	80.71±10.19	114.07±12.70**	73.55±10.48^##
ALT	39.35±11.69	47.36±11.37	32.16±3.96^##
AST/ALT	2.16±0.47	2.52±0.59	2.30±0.32
HDL-C (mmol/L)	2.30±0.54	4.23±0.96**	4.40±0.66**
LDL-C (mmol/L)	0.31±0.06	0.91±0.20**	0.53±0.06**^##
TC (mmol/L)	3.26±0.65	7.29±1.18**	5.85±0.54**^##
AI	0.44±0.27	0.79±0.41	0.35±0.20^#

Fig.3 Effect of DWYG Decoction on contents of Lyso-GPLs in circulating exosomes in NAFLD mice. A: Relative peak areas of blood circulation exosome lysophospholipids in the normal, model and treatment group. B: PCA score plots of normal, model and DWYG groups.

Fig.4 Results of OPLS-DA analysis of the samples in the 3 groups. A: OPLS-DA analysis of normal and model groups. B: 999-time permutation validation results of the normal and model groups. C: OPLS-DA analysis of the treatment and model groups. D: 999-time permutation validation results of the treatment and model group.

Fig.5 Relative contents of 43 Lyso-GPLs. A: Fold change. B: Relative content in normal, model and compound groups.

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