Effects of Desmodium caudatum on Gastrointestinal Hormones and Intestinal Flora in Rats with Gastritis

Summary

The present protocol describes the method for analyzing intestinal flora using Illumina-based 16S rRNA gene sequencing and provides a framework for evaluating the effectiveness of herbal decoctions.

Abstract

In order to preliminarily explore the effects of Desmodium caudatum on gastritis and intestinal flora in rats, a chronic gastritis rat model was established using the classic sodium salicylate method. Eighteen SPF rats were divided into three groups: the control group (Group C), the model group (Group M), and the treatment group (Group T). Pathological sections of the gastric wall were taken from rats in each group. Furthermore, the concentrations of gastrin and malondialdehyde in the serum of rats in each group were determined by ELISA. Additionally, the effects of D. caudatum on the intestinal flora of rats with gastritis were explored through a detailed comparison of gut bacterial communities in the three groups, employing Illumina-based 16S rRNA gene sequencing. The results indicated that D. caudatum decoction could reduce the malondialdehyde content and increase the gastrin content. Moreover, D. caudatum decoction was found to enhance the diversity and abundance of intestinal flora, exerting a positive impact on the treatment of gastritis by regulating and restoring the intestinal flora.

Introduction

Chronic gastritis (CG), one of the most common clinical diseases, is characterized by chronic and persistent inflammatory changes in the gastric mucosa epithelium, which is frequently and repeatedly attacked by various pathogenic factors¹. The incidence rate of CG ranks first among all types of stomach diseases, accounting for 40% to 60% of the outpatient service rate in the Department of Gastroenterology². Moreover, the incidence rate generally increases with age, especially in individuals who are middle-aged and older³. Undoubtedly, CG significantly reduces people's quality of life, emphasiz....

Protocol

The procedures for the care and use of animals were approved by the Ethics Committee of Mianyang Normal University, and all relevant institutional and governmental regulations concerning the ethical use of animals were strictly followed. For the present study, SPF rats (Kunming species, both male and female, weighing 180-220 g) were utilized. The animals were obtained from a commercial source (see Table of Materials). All animals were housed in a pathogen-free environment and provided ad libitum.......

Discussion

D. caudatum, a commonly used folk medicine by the Qiang nationality¹², has shown significant efficacy in treating gastrointestinal diseases. With the advancement of modern pharmacological research, the imbalance of flora resulting from gastrointestinal microecology imbalance is identified as a key factor in acute and chronic gastrointestinal diseases^22,23. Certain microorganisms in the intestinal tract play a crucial role in maint.......

Materials

Name	Company	Catalog Number	Comments
Alpha diversity analysis	Mothur 1.30.2
AxyPrep deoxyribonucleic acid (DNA) gel extraction kit	Axygen Biosciences	AP-GX-50
Beta diversity analysis	Qiime 1.9.1
Cryogenic refrigerator	Forma-86C ULT freezer
Desmodium caudatum	The Traditional Chinese Medicine Hospital of Beichuan Qiang Autonomous County
E.Z.N.A. soil kit	Omega Bio-tek	D5625-01
Illumina MiSeq Platform	Illumina Miseq PE300/NovaSeq PE250
Multiskan Spectrum	spectraMax i3
OTU clustering	Uparse 7.0.1090
OTU statistics	Usearch 7.0
PCR instrument	TransGen AP221-02
PCR instrument	ABI GeneAmp 9700
QuantiFluor-ST double-stranded DNA (dsDNA) system	Promega Corp.
Sequence classification annotation	RDP Classifier 2.11
Sodium salicylate	Sichuan Xilong Chemical Co., Ltd	54-21-7
SPF rats	Chengdu Dashuo Experimental Animal Co., Ltd
SPSS 18.0	IBM