Traditional herbal formula Wu-Mei-Wan alleviates TNBS-induced colitis in mice by inhibiting necroptosis through increasing RIPK3 O-GlcNAcylation

Abstract

Background Accumulating evidence indicated that necroptosis plays an essential role in the pathogenesis of inflammatory bowel disease (IBD). The O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) of necroptotic signal molecule receptor-interacting serine-threonine kinase 3 (RIPK3) was reported to exert a protective effect in gut inflammation. Our recent study suggested traditional Chinese herbal formula Wu-Mei-Wan (WMW) as an effective prescription in mouse colitis. However, the potential mechanisms are not fully understood. Considering the crucial role of necroptosis in the pathogenesis of IBD, therefore, this study was designed to explain whether the anti-colitis effect of WMW is mediated by modulating necroptosis and its related mechanisms. Methods The protective effects of WMW on colitis have been determined by detecting colitis mice body weight, disease activity index (DAI), survival rate and colon length. Colonic inflammation was examined by inflammatory cells infiltration and local cytokines levels. After then, we measured the levels of necroptosis and O-GlcNAcylation. C O-immunoprecipitation experiments were used to address whether elevated O-GlcNAcylation can inhibit necroptotic signal transduction in the treatment of WMW. Finally, the key enzymes in O-GlcNAcylation: O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) were examined and molecular docking analysis was used to determine effective natural compounds in the regulation on OGT and OGA activities. Results Our results showed that WMW significantly improved mice body weight, survival rate and colon length, decreased DAI in TNBS-induced colitis. WMW obviously alleviated colonic inflammatory responses with reduced macrophages, neutrophils infiltration and local IL-1β, IL-6, TNF-α and IFN-γ levels. It was found that WMW increased colonic O-GlcNAcylation level and inhibited the activation of RIPK1, RIPK3 and MLKL. Then, further experiments revealed that WMW enhanced OGT activity and suppressed OGA activity, thereby increasing RIPK3 O-GlcNAcylation and inhibiting the binding of RIPK3 and MLKL, which led to the inhibition of necroptosis. Additionally, docking analysis demonstrated that hesperidin, coptisine and ginsenoside Rb1 may exert a major role in the regulation on OGT and OGA activities by WMW. Conclusion Our work demonstrated that WMW can alleviate TNBS-induced colitis in mice by inhibiting necroptosis through increasing RIPK3 O-GlcNAcylation.

Targeting ferroptosis in acute kidney injury

“…Cells undergoing necroptosis exhibit disrupted cell membranes, which is the major difference from necrosis. The RIPK3 and its substrate MLKL play critical roles in the formation of necroptosis [ 99 – 101 ]. In acute ischemic kidney injury, the expression of acyl-CoA synthetase long-chain family member 4 (ACSL4, a key enzyme in ferroptosis) is increased and is correlated with the severity of renal damage [ 82 , 102 ].…”

Wumei Pill Ameliorates AOM/DSS-Induced Colitis-Associated Colon Cancer through Inhibition of Inflammation and Oxidative Stress by Regulating S-Adenosylhomocysteine Hydrolase- (AHCY-) Mediated Hedgehog Signaling in Mice

“…As a traditional Chinese medicine (TCM), WMP is widely used to treat digestive system diseases in clinical practice in China [40]. Previous studies showed that WMP had extensive pharmacological effects, including alleviated trinitrobenzenesulfonic acid-(TNBS-) induced colitis [41], prevented high-fat diet-induced obesity [42], protected pancreatic β cells [43], and reduced insulin resistance [44]. Noteworthy, WMP can treat roundworm coldness in the extremities and longtime diarrhea of cold-heat complex type based on the "Treatise on Febrile Disease."…”

“…Although there is no record of CAC in TCM literatures, however, according to its clinical characteristics and pathogenesis, CAC can be classified as a category of "dysentery" and "risceral intoxication." WMP is a famous formula, which is composed of ten herbs: Fructus Mume, Herba Asari, Rhizoma Zingiberis, Rhizoma Coptidis, Radix Angelicae sinensis, Rhizoma Typhonii gigantei, Pericarpium Zanthoxyli, Ramulus cinnamomi, Radix Ginseng, and Cortex Phellodendri [41]. Many preclinical and clinical trials found that WMP was suitable for the early prevention and treatment of inflammatory bowel disease and CAC.…”

Wu-Mei-Wan Ameliorates Murine Ulcerative Colitis by Regulating Macrophage Polarization

“…WMW, a traditional medicine in China, has been used to treat chronic intestinal inflammatory disorders for two millennia. Recent studies have found that WMW significantly reduces the acute colonic inflammatory response induced by trinitrobenzenesulfonic acid by reducing the infiltration of macrophages and neutrophils and the levels of local IL-1 β , IL-6, TNF- α , and IFN- γ ( Wu F. et al, 2021 ). UC is a chronic inflammatory disorder, and DSS is commonly used to induce UC in animal models.…”

“…An increasing body of evidence shows that WMW has anti-colon necrosis and anti-intestinal fibrosis effects and can inhibit intestinal inflammation and repair damaged intestinal mucosa. However, the specific effector mechanism(s) of WMW in the treatment of UC are not clear ( Wu et al, 2020a ; Jiang et al, 2020 ; Wu F. et al, 2021 ). WMW is made from 10 plants, including smoked plum, wild ginger (Asarum sieboldii), ginger (Zingiber officinale), Aconitum carmichaelii, Zanthoxylum, Cinnamomum cassia, Coptis chinensis, Phellodendron amurense, Panax ginseng, and Angelica sinensis.…”