Evaluating the Mechanism of a Traditional Herbal Syrup for Anemia Treatment Using Liver Metabolomics

Summary

This study primarily introduces the application of liver metabolomics in investigating the effectiveness of Shi-Liu-Bu-Xue Syrup in treating anemia.

Abstract

As a well-known Uyghur medicine, Shi-Liu-Bu-Xue Syrup (SLBXS) has been widely used to treat anemia in China for over 20 years. However, the underlying mechanisms of its effectiveness in treating anemia remain unclear. In this study, liver metabolomics was primarily employed to determine the potential regulatory mechanisms of SLBXS in treating anemia. Liver metabolomics profiling was conducted to characterize the mechanism of action of SLBXS in an acetylphenylhydrazine-induced mouse model of anemia. SLBXS was shown to decrease liver index, white blood cell count, and platelet count, while increasing red blood cell count, hemoglobin, and hematocrit levels. Core targets were selected for verification using Western blotting. SLBXS demonstrated a significant therapeutic effect on anemia primarily by regulating galactose metabolism and the HIF-1 signaling pathway, as indicated by the downregulation of HIF-1α, NOS3, VEGFA, and GLA proteins in the liver tissues of anemic mice. This study clarifies the potential regulatory mechanisms of hepatic metabolism by SLBXS administration in treating anemia.

Introduction

Anemia is a pressing and prevalent global health issue, affecting 25% of the world's population and people of all ages, particularly adolescents and pregnant women^1,2,3. It is associated with an increased risk of preterm labor and maternal mortality and can lead to physical developmental disorders and impaired cardiovascular performance⁴. This condition may also negatively impact the health status of adolescents, resulting in infections and heart failure⁵. Current treatments primarily include blood tran....

Protocol

All experimental procedures were approved by the Laboratory Animal Ethics Committee of the Hubei University of Chinese Medicine (HBUCMS201912015). Male C57BL/6 mice (weight 20-22 g) were housed in a specific pathogen-free room with a relative humidity of 50%-60% and a temperature of 22 °C ± 2 °C, subjected to a 12 h light/12 h dark cycle, and provided with free access to food and water. Before the experiment began, all mice were allowed one week to acclimate to the environment. Mice were randomly assigned .......

Discussion

Anemia is a common condition affecting many people worldwide, particularly in developing countries¹. In China, patients frequently use traditional Chinese medicine, including Uyghur medicine, to alleviate the signs and symptoms of anemia. SLBXS is a Uyghur medicine that has been used in clinical practice for many years; however, its exact mechanism of action against anemia remains poorly understood¹³. In this study, a mouse model of acetylphenylhydrazine (APH)-induced anemi.......

Materials

Name	Company	Catalog Number	Comments
Acetylphenylhydrazine	Shanghai Aladdin Biochemical Technology Co., Ltd.	C13979660
Automatic chemiluminescence imaging analysis system	Shanghai Tanon Life Science Co., Ltd.	Tanon-5200
Bicinchoninic acid assay kit	ThermoFisher Scientific	QPBCA-1KT
Capillary column	Agilent J&W Scientific, Agilent Technologies, Inc.	DB-5MS
Cell lysis buffer for Western and IP	Beyotime Biotechnology	P0013
Chlorotrimethylsilane	Shanghai Aladdin Biochemical Technology Co., Ltd.	C104814
Electronic balance	Mettler-Toledo International Inc.	ME203E
Electronic scale	Mettler-Toledo International Inc.	LE104E
Fu-Fang-E-Jiao Syrup	Dong E E Jiao Co., Ltd.	214020031
Fully automatic hemocyte analyzer	Shenzhen Mindray Animal Care Technology Co., Ltd.	IDEXX ProCyte Dx
GC-MS system	Agilent Technologies, Inc.	7890B-5977B
GLA primary antibody	Bioworld Technology	BS77041
Glass homogenizer	Shanghai Lei Gu Instruments Co., Ltd.	B-013001
Glass rod	Shanghai Lei Gu Instruments Co., Ltd.	B-003904
GraphPad Prism software	GraphPad, La Jolla	Version 9.0
Heparinized sample tubes	Changde BKMAM Biotechnology Co., Ltd.	B-ACT1P5
HIF-1α primary antibody	Bioworld Technology	BS3514
HMDB database	http://www.hmdb.ca/	—
Isoflurane	Hebei Jindafu Pharmaceutical Co., Ltd.	20231202
Male C57BL/6 mice	Liaoning Changsheng Biotechnology Co., Ltd.	No. SCXK [Liao] 2015-0001
MassHunter	Agilent Technologies, Inc.	B.08.00
MetaboAnalyst 5.0	https://www.metaboanalyst.ca/	—
Methoxyamine hydrochloride	Shanghai Aladdin Biochemical Technology Co., Ltd.	E1818113
n-hexane	Shanghai Aladdin Biochemical Technology Co., Ltd.	C14878803
NIST database	http://webbook.nist.gov/chemistry/	—
NOS3 primary antibody	Bioworld Technology	BS3625
Pyridine	Shanghai Aladdin Biochemical Technology Co., Ltd.	C13026996
Saline	BIOSHARP LIFE SCIENCES	2308262009
Shi-Liu-Bu-Xue Syrup	Xinjiang Uygur Pharmaceutical Co., Ltd.	211277
Surgical manipulation plate	DIXSG	ZK-JPB-A
VEGFA primary antibody	Bioworld Technology	AP0742
β-actin	ABclonal (Shanghai) Trading Co., Ltd.	AC026