# Sanhuang Xiexin decoction ameliorates DSS-induced colitis in mice by regulating intestinal inflammation, intestinal barrier, and intestinal flora (2023)

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Volume 297,

28 October 2022

, 115537

## Abstract

### Ethnopharmacological relevance

Sanhuang Xiexin decoction (SXD) is a widely applicated traditional Chinese medicine (TCM) with a significant intestinal anti-inflammatory effect.

### Aim of the study

To evaluate the therapeutic effect and elucidate the possible underlying mechanisms of SXD on dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice.

### Methods

To model UC, 3% DSS was added to the drinking water for 7 days. The UC mice were grouped and treated with three doses of SXD (1.3, 2.6, and 6g/kg) orally for 7 days. Mice body weight and disease activity index (DAI) scores were recorded daily. After treatment with SXD, the colon was removed, and the colon length and histopathological changes were recorded. Blood cells were counted and colonic inflammatory cytokines and oxidative stress indicators were examined. The key proteins in TLR4-MyD88-NF-κB signaling and the colonic barrier were determined by Western blot analysis. The restorative effect of SXD on intestinal flora was determined.

### Results

Treatment with SXD reduced DAI scores, increased body weight, improved colon shortening, and decreased colonic damage. SXD decreased the numbers of white blood cells (WBCs), increased the numbers of red blood cells (RBCs), and inhibited the expression of inflammatory cytokines and oxidative stress indicators. In addition, SXD displayed an effective anti-inflammatory effect by inhibiting the expression levels of p-IκBα, TLR4, MyD88, and p65. Furthermore, SXD significantly restored the integrity of the colonic barrier and the abundance of beneficial flora.

### Conclusions

SXD significantly reduced DSS-induced colon damage when the dose was higher than 1.3g/kg, and the middle dose group (2.6g/kg) indicated the best effect. SXD effectively ameliorated DSS-induced UC in mice, possibly by inhibiting oxidative stress, protecting the mucosal barrier, inhibiting the TLR4-MyD88-NF-κB signaling pathway, and regulating the intestinal flora.

## Introduction

Ulcerative colitis (UC) is the consequence of inflammatory bowel disease, which is generally characterized by nonspecific, persistent, and chronic intestinal ulcers. The disease is persistent and difficult to heal and has the risk of becoming malignant. It has been considered a stubborn modern disease by the World Health Organization (Pei et al., 2019). The incidence and prevalence of UC are increasing worldwide (Molodecky et al., 2012). Diarrhea, mucus pus, bloody stools, and abdominal pain are typical symptoms of UC (Jdf et al., 2019). The disease pathogenesis of UC is still not fully understood, but accumulating research has indicated that colitis occurs due to a mixture of genetics, autoimmune diseases, changes in the intestinal flora, and environmental factors (Ng et al., 2012). The risk factors of UC, including infection, medication, diet, and tobacco use, play important roles in the disruption of the intestinal mucosa barriers (Du and Ha, 2020). The host's innate and adaptive immunity can prevent the invasion of harmful bacteria and the disruption of normal intestinal microbiota. Many recent experiments confirmed that the imbalance in the intestinal microbial immune response was closely related to the occurrence, progression, and changes of UC (Zheng et al., 2017).

Anti-inflammatory drugs such as aminosalicylic acid and steroid hormones are used to treat UC. However, the pathogenesis of UC has not been fully elucidated and these drugs only alleviate some symptoms of UC and have obvious side effects (Currò et al., 2017; Rosenberg and Peppercorn, 2010). Therefore, it is of great significance to continue to find more effective and less toxic drugs for the treatment of UC. Traditional Chinese medicine (TCM) and formulations are multi-component, multi-targeted, and multi-channeled. Compared with single chemical drugs, TCM, with complex ingredients and mechanisms, has certain advantages in the treatment of UC. According to the basic theory of etiology treated by TCM, damp-heat plays a leading role in the pathogenesis of UC. In the treatment of UC, clearing away heat and drying dampness are regular therapeutic principles (Shen et al., 2020). The formulations consisting of Kushen, Huangqin, and Huanglian, with the effect of clearing away heat and dampness, showed good efficacy in treating UC (An et al., 2020; Chen et al., 2020).

Sanhuang Xiexin decoction (SXD) is another classic heat and dampness-clearing prescription, from the “Synopsis of Prescriptions of the Golden Chamber” written by Zhang Zhongjing in the Eastern Han Dynasty. According to the theory of TCM, SXD has the effects of clearing heat, purging fire, detoxifying, and hemostasis(CHEN, 2016). SXD consists of three traditional herbals, Rhei Radix et Rhizoma, Coptidis Rhizoma, and Scutellariae Radix, at a ratio of 2:1:1. SXD is widely used in the treatment of intestinal diseases and demonstrated antibacterial, anti-inflammatory, and hemostatic activity in both preclinical studies and clinical applications (Han et al., 2013; Lo et al., 2005). Scutellariae Radix even has adaptogenic activity (Panossian et al., 2021).Currently, Rhei Radix et Rhizoma, Coptidis Rhizoma, Scutellariae Radix and related formulas are also widely used in Japan and Korea (Choi et al., 2013; Hirayama et al., 2018; Kwon et al., 2016). Rheum palmatum L. is a medicinal plant in Russia and included in the State Pharmacopoeia of Russia 14th edition(Shikov et al., 2021). It is also widely used in European Union and there is Community herbal monograph on Rhei radix (Rheum palmatum L./Rheum officinale Baillon)(Galen.2014). Studies showed that SXD contains a large number of active components, including berberine, coptisine, baicalin, baicalein, rhein, and others (Li et al., 2010). These components have been widely studied and their activities are related to anti-inflammatory, antioxidant, and antibacterial effects, as well as the regulation of intestinal flora (Dong et al., 2022; Wu et al., 2019; Zhu et al., 2019, 2020). Therefore, SXD showed great therapeutic potential for UC. However, the therapeutic effect and mechanism of SXD in treating UC have not been fully evaluated.

In this paper, the therapeutic effect of SXD was evaluated in mice with dextran sulfate sodium (DSS)-induced UC. In addition, the anti-UC mechanism of SXD was also explored by assessing changes in oxidative stress, the mucosal barrier, inflammation, and intestinal flora in SXD-treated mice.

## Animals

Four-week-old C57BL/6 mice (18±2g, n=72) were obtained from Chengdu Dossy Experimental Animals Co., Ltd. (Chengdu, China) (Certificate No. SCXK (Chuan) 2020–030). All mice were fed in the animal laboratory, and the environmental temperature and humidity were kept at 25°C±2°C and 50%±5%, respectively, with a 12h light/dark cycle. The mice had free access to standard water and food. The in vivo experiments were performed under guidelines that were approved by the Institutional Animal

## SXD attenuates symptoms of DSS-induced UC mice

Three percent DSS significantly induced diarrhea, hematochezia, and weight loss in mice in the UC group (Fig. 1A). The DAI score of the UC group was significantly increased compared with control group (Fig. 1B). Diarrhea and hematochezia in mice in the SXD groups and SASP group were less severe compared with UC group. Therefore, SXD could significantly decrease DAI scores in UC mice (Fig. 1B). As shown in Fig. 1 C and D, there was a significant shortening of the colon length in the UC group,

## Discussion

SXD is a classic TCM formula with a history of more than 1000 years in clinical application. The composition of TCM compounds is very complex, and not all components have pharmacological activity. Their therapeutic effect is the result of the comprehensive action of some components with pharmacological activity (Yuan et al., 2020). In this experiment, SXD decoction was first extracted according to the traditional decocting method, and 13 main active components in SXD were determined according

## Conclusion

SXD significantly alleviated clinical symptoms, inflammation, and colonic damage of DSS-induced UC in mice. The recommended dose is 2.6g/kg once a day for treating DSS-induced UC. The mechanism may be related to the inhibition of oxidative stress, protection of the mucosal barrier, inhibition of the TLR4-MyD88-NF-κB signaling pathway, and regulation of intestinal flora.

## CRediT authorship contribution statement

Xueyuan Wu: Conceptualization, Methodology, Writing – original draft. Shu Fu: Conceptualization, Methodology. Miao Jiang: Conceptualization, Methodology, Software. Jing Wang: Data curation. Huaqiao Tang: Data curation, Writing – original draft. Chunlin Fang: Software, Data curation. Wen Li: Supervision, Writing – review & editing. Chaomei Fu: Supervision, Writing – review & editing.

## Declaration of competing interest

The authors declare no conflict of interest related to this paper.

## Acknowledgments

This work was supported by the National Natural Science Foundation of China (81903812).

## References (71)

• L. Zhu et al.Protective effects of berberine hydrochloride on DSS-induced ulcerative colitis in rats

### Int. Immunopharm.

(2019)

• X.-J. Zhang et al.Palmatine ameliorated murine colitis by suppressing tryptophan metabolism and regulating gut microbiota

### Pharmacol. Res.

(2018)

• Z. Yuan et al.Therapeutic effect of n-butanol fraction of Huang-lian-Jie-du Decoction on ulcerative colitis and its regulation on intestinal flora in colitis mice

### Biomed. Pharmacother.

(2020)

• Y. Xia et al.Lactobacillus plantarum AR113 alleviates DSS-induced colitis by regulating the TLR4/MyD88/NF-κB pathway and gut microbiota composition

### J. Funct.Foods

(2020)

• S.-C. Wu et al.Baicalin protects mice against Salmonella typhimurium infection via the modulation of both bacterial virulence and host response

### Phytomedicine

(2018)

• M. Tillhon et al.Berberine: new perspectives for old remedies

### Biochem. Pharmacol.

(2012)

• N.-R. Shin et al.Proteobacteria: microbial signature of dysbiosis in gut microbiota

### Trends Biotechnol.

(2015)

• A.N. Shikov et al.Medicinal plants from the 14th edition of the Russian Pharmacopoeia, recent updates

### J. Ethnopharmacol.

(2021)

• V. Rani et al.Oxidative stress and metabolic disorders: pathogenesis and therapeutic strategies

### Life Sci.

(2016)

• G.P. Ramos et al.Mechanisms of Disease: Inflammatory Bowel Diseases, Mayo Clinic Proceedings

(2019)

• Y. Lo et al.San-Huang-Xie-Xin-Tang attenuates inflammatory responses in lipopolysaccharide-exposed rat lungs

### J. Ethnopharmacol.

(2005)

• Y. Liu et al.Berberine inhibits macrophage M1 polarization via AKT1/SOCS1/NF-κB signaling pathway to protect against DSS-induced colitis

### Int. Immunopharm.

(2018)

• W. Liu et al.Andrographolide sulfonate ameliorates experimental colitis in mice by inhibiting Th1/Th17 response

### Int. Immunopharm.

(2014)

• S.-L. Li et al.UPLC–PDA–TOFMS based chemical profiling approach to rapidly evaluate chemical consistency between traditional and dispensing granule decoctions of traditional medicine combinatorial formulae

### J. Pharmaceut. Biomed. Anal.

(2010)

• L. Jialing et al.Changes in serum inflammatory cytokine levels and intestinal flora in a self-healing dextran sodium sulfate-induced ulcerative colitis murine model

### Life Sci.

(2020)

• I.I. Ivanov et al.Intestinal commensal microbes as immune modulators

### Cell Host Microbe

(2012)

• X.-H. Han et al.Relationships between pharmacokinetics and efficacy of Xie-xin decoction in rats with experimental ulcerative colitis

### J. Ethnopharmacol.

(2013)

• L. Du et al.Epidemiology and pathogenesis of ulcerative colitis

### Gastroenterol. Clin.

(2020)

• Y.Y. Choi et al.Inhibitory effect of Coptis chinensis on inflammation in LPS-induced endotoxemia

(2009)

• N. An et al.

### Med.

(2020)

• J.W. Baynes et al.

### Diabetes

(1999)

• D. Boltin et al.

### J. Clin. Gastroenterol.

(2013)

• B. Chassaing et al.

### Curr. Protoc. Im.

(2014)

• M. Chen et al.

(2020)

• Z.-J. Chen

### Research status of Sanhuang Xiexin Decoction and analyses of key technologies and core issues

(2016)

• H.S. Cooper et al.

### Lab. Investigation J. Tech. Method Pathol.

(1993)

• S. Coufal et al.

### Cells

(2019)

• D. Currò et al.

### Front. Pharmacol.

(2017)

• I. D'Odorico et al.

### J. Digestive Dis.

(2018)

• L. Dong et al.

### Phytother Res.

(2022)

• D.D. Eichele et al.

(2017)

• E. Galen

### J. Ethnopharmacol.

(2014)

• M.V. Gulubova et al.

### Int. J. Colorectal Dis.

(2007)

• S. Guo et al.

(2021)

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