Gegen Qinlian decoction maintains colonic mucosal homeostasis in acute/chronic ulcerative colitis via bidirectionally modulating dysregulated Notch signaling
Graphical abstract
Introduction
Studies have demonstrated that the secretion of trefoil peptides and mucine glycopeptides by goblet cells is essential in the initial phase (minutes to several days) of intestinal mucosal repair progress, termed “restitution” (Podolsky, 1997). The replacement of lost cells through proliferation and differentiation is crucial in the “second phase” of intestinal restoration (Nusrat et al., 1992). However, in some clinical conditions, such as ulcerative colitis (UC), the homeostasis and regeneration of intestinal epithelia is severely disturbed and present persistent mucosal ulceration on the inner lining of the large intestine and spontaneously replasing inflammation (Algieri et al., 2014; Hernandez-Chirlaque et al., 2016). In view of this, promoting intestinal regeneration and restoring intestinal mucosal homeostasis is an effective strategy for the treatment of UC. (Okamoto and Watanabe, 2004).
Crypt base columnar (CBC) stem cells are responsible for the regeneration of intestinal tissue (Barker et al., 2007). Replicating colonic CBC stem cells can self-renew or produce rapidly proliferating transit-amplifying cells that differentiate into two major differentiated cell types: absorptive colonocytes and mucus-secreting goblet cells (Leushacke and Barker, 2014). Notch signaling controls the fate of CBC stem cells and critically influences absorptive versus secretory cell fate decisions in the intestinal epithelium (Noah and Shroyer, 2013). The Notch pathway is a conserved mechanism of cell-cell interaction that regulates cell differentiation, proliferation, survival and development (Ortiz-Masia et al., 2016). In mammalian cells, ligation of Notch receptors to ligands leads to proteolytic cleavage of Notch by an intracellular γ-secretase complex and releases the Notch intracellular domain (NICD). Then, the NICD translocates to the nucleus and binds to DNA-binding protein recombinant-recognition-sequence-binding protein at the Jκ site (RBP-J) (Hu et al., 2008; Wang et al., 2017). This interaction recruits other co-factor proteins of the mastermind-like (MAML) family, and the RBP-J–NICD–MAML complex affects the activation of Notch target gene, hairy and enhancer of split (Hes) (Boriushkin et al., 2019; Wang et al., 2017). Hes is a crucial mediator of the effect of Notch on cell fate determination, and activation of Notch signaling increases Hes1 expression accompanied by augmented proliferation of CBC stem cells and repressed differentiation of secretory goblet cells in the extended proliferation zone, whereas disruption of Notch signaling decreases Hes1 expression and activates mouse atonal homoleg 1 (Math1) transcription coupled with a loss of CBC stem cell proliferation and secretory cell hyperplasia (Okamoto and Watanabe, 2004; Pellegrinet et al., 2011; VanDussen et al., 2012). Therefore, homeostasis and regeneration of the intestine is abrogated when Notch signaling is disrupted.
Research has shown that traditional Chinese medicine (TCM) is a source of pharmaceutical material that can be used in the treatment of various diseases with fewer associated adverse effects (Zhang et al., 2019). Gegen Qinlian decoction (GQ) composed of four quality-assured herbs, Scutellaria baicalensis Georgi., Coptis chinensis Franch., Pueraria lobata (Willd.) Ohwi. and Glycyrrihiza uralensis Fisch., has been used as an efficacious medicine against acute diarrhea for approximately 2000 years and has recently been clinically proven effective in the treatment of UC. In our previous study, we reported that GQ alleviated the hyperactive immune response by suppressing TLR4/NF-κB signaling and enhancing the antioxidant effect in acute colitis mice (Li et al., 2016a). To further explore the multiple targets and therapeutic mechanism of GQ, in this study, we focused on the role of GQ in the regeneration of the colonic mucosa through restoring dysregulated Notch signaling in DSS-induced acute and chronic UC mice (Algieri et al., 2014; Koboziev et al., 2011). We observed that Hes1 expression was upregulated in acute UC mice and was downregulated in chronic UC mice. Therefore, we hypothesized that GQ exerts its homeostatic effect on the colonic mucosa by modulating dysregulated Notch signaling in UC mice.
Section snippets
Reagents
DSS (36–50 kDa) was purchased from MP Biomedicals (California, USA). Antibodies against Hes1 (1:1000), MAML (1:1000) and Toll-like receptor 4 (TLR4) (1:1000) were purchased from Cell Signaling Technology (Colorado, USA), RBP-J (1:1000), Math1 (1:1000), Glyceraldehyde phosphate dehydrogenase (GAPDH) (1:5000) and β-actin (1:5000) antibodies were obtained from ABclonal Biotech Co., Ltd (Wuhan, China). Lipopolysaccaride (LPS) was purchased from Sigma-Aldrich (Missouri, USA). Tumor necrosis factor α
Quality control of GQ
The HPLC fingerprint of GQ at a wavelength of 270 nm is shown in Fig. 1. Six components in GQ were identified and quantified based on the retention times and peak areas with those of the reference standards.
GQ repaired the colonic mucosa through downregulation of Hes1 expression in acute UC mice
The acute UC mice were improved by GQ treatment, as shown in Figs. S3 and 2F. The weight loss of the mice in the L-GQ, H-GQ and BBR groups was ameliorated by administration of different doses of GQ and BBR (Fig. S3B). On the 12th day, the body weight of the mice in the L-GQ, H-GQ and BBR
Discussion
As an effective TCM prescription for treating diarrhea, GQ alleviates intestinal disorders by modulating the gut microbiota, reducing excessive inflammation and repairing the intestinal mucosa (Liu et al., 2019a,b; Lv et al., 2019; Wu and Tan, 2019; Wu et al., 2019; Xu et al., 2015). However, the molecular mechanism by which GQ repairs the damaged intestinal mucosa remains unclear. Notch signaling is crucial in regulating both the differentiation and proliferation of intestinal epithelial cells
Conclusion
In conclusion, our results revealed for the first time that GQ restored the regeneration and homeostasis of the colonic mucosa via bidirectionally modulating dysregulated Notch signaling in acute/chronic UC models. GQ enhanced hypoactive Notch signaling in chronic UC mice and promoted the proliferation of the epithelium and differentiation of absorption cell lines, while GQ suppressed hyperactive Notch signaling in acute UC mice to promote the differentiation of goblet cells and secretion of
CRediT authorship contribution statement
Yaxing Zhao: Conceptualization, Funding acquisition, Data curation, Formal analysis, Writing - review & editing. Haofan Luan: Funding acquisition, Data curation, Formal analysis, Writing - review & editing. Hui Gao: Funding acquisition, Writing - review & editing. Xiaojun Wu: Writing - review & editing. Yubin Zhang: Conceptualization, Writing - review & editing. Ruiyan Li: Conceptualization, Writing - original draft, Writing - review & editing, Data curation, Formal analysis.
Declaration of Competing Interest
The authors have no competing interests.
Acknowledgment
This work was supported by funding from the Opening Project of Shanghai Key Laboratory of Compound Chinese Medicines (Shanghai University of Traditional Chinese Medicine) (No. 17DZ2273300) and the Basic Research Project of China Pharmaceutical University (No. 2632018PY06) to Ruiyan Li; and the National Natural Science Foundation of China (No. 81573484) to Yubin Zhang.
References (36)
- et al.
Intestinal anti-inflammatory activity of the Serpylli herba extract in experimental models of rodent colitis
J. Crohns Colitis
(2014) - et al.
Proinflammatory signaling regulates hematopoietic stem cell emergence
Cell
(2014) - et al.
Intervention effects of QRZSLXF, a Chinese medicinal herb recipe, on the DOR-beta-arrestin1-Bcl2 signal transduction pathway in a rat model of ulcerative colitis
J. Ethnopharmacol.
(2014) - et al.
Inflammatory signaling regulates hematopoietic stem and progenitor cell emergence in vertebrates
Blood
(2015) - et al.
Integrated regulation of Toll-like receptor responses by Notch and interferon-gamma pathways
Immunity
(2008) - et al.
Gegen Qinlian decoction alleviates experimental colitis via suppressing TLR4/NF-KappaB signaling and enhancing antioxidant effect
Phytomedicine
(2016) - et al.
Berberine ameliorates chronic relapsing dextran sulfate sodium-induced colitis in C57BL/6 mice by suppressing Th17 responses
Pharmacol. Res.
(2016) - et al.
Gegen Qinlian decoction treats diarrhea in piglets by modulating gut microbiota and short-chain fatty acids
Front. Microbiol.
(2019) - et al.
Preventive and therapeutic oral administration of the pentacyclic triterpene alpha,beta-amyrin ameliorates dextran sulfate sodium-induced colitis in mice: the relevance of cannabinoid system
Mol. Immunol.
(2013) - et al.
Baicalin protects mice against Salmonella typhimurium infection via the modulation of both bacterial virulence and host response
Phytomedicine
(2018)
Interaction between gut microbiota and ethnomedicine constituents
Nat. Prod. Rep.
Traditional Chinese medicine Gegen Qinlian decoction ameliorates irinotecan chemotherapy-induced gut toxicity in mice
Biomed. Pharmacother.
Comparative comprehension on the anti-rheumatic Chinese herbal medicine Siegesbeckiae Herba: combined computational predictions and experimental investigations
J. Ethnopharmacol.
Distinct cytokine patterns identified from multiplex profiles of murine DSS and TNBS-induced colitis
Inflamm. Bowel Dis.
Identification of stem cells in small intestine and colon by marker gene LGR5
Nature
The Notch signaling pathway: transcriptional regulation at Notch target genes
Cell Mol. Life Sci.
Kruppel-like factor 4 regulates developmental angiogenesis through disruption of the RBP-J-NICD-MAML complex in intron 3 of DLL4
Angiogenesis
Germ-free and antibiotic-treated mice are highly susceptible to epithelial injury in DSS colitis
J. Crohns Colitis
Cited by (52)
Herbal medicine for Hashimoto's thyroiditis: A systematic review and network meta-analysis
2024, Journal of EthnopharmacologyA new rat model of chronic relapsing ulcerative colitis with traditional Chinese medicine syndrome of yang deficiency and dampness stagnation
2023, Pharmacological Research - Modern Chinese MedicineGegen Qinlian decoction activates AhR/IL-22 to repair intestinal barrier by modulating gut microbiota-related tryptophan metabolism in ulcerative colitis mice
2023, Journal of EthnopharmacologyCitation Excerpt :By analyzing and summarizing the clinical trial literature of GQD in the treatment of UC published in common databases, the results showed that the average total effective rate of GQD in the treatment of UC was 92.4%, and the average cure rate was 47.2% (Wang et al., 2022). Other studies confirmed that GQD is effective against inflammatory intestinal diseases, including diarrhoea, ulcerative colitis, and intestinal adverse reactions caused by chemotherapeutic agents (Liu et al., 2019; Zhao et al., 2020). Animal studies have confirmed that GQD alleviates colitis in mice by repairing the damaged intestinal barrier and regulating the gut microbiota (Li et al., 2020; Wang et al., 2021), suggesting that there may be a relationship between these two components in the GQD treatment of UC.
Demethyleneberberine blocked the maturation of IL-1β in inflammation by inhibiting TLR4-mitochondria signaling
2022, International ImmunopharmacologyBidirectional effects and mechanisms of traditional Chinese medicine
2022, Journal of Ethnopharmacology