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Moxibustion Inhibits the Expression of Colonic NLRP3 through miR7/RNF183/NF-κB Signaling Pathway in UC Rats
Xi-Ying Li1, Yan-Ting Yang2, Yue Zhao2
1Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Jingan District, Shanghai 200071, China.
Background:
Moxibustion has been recognized as an effective approach for ulcerative colitis, yet its mechanism is not clear. The research aimed to investigate the influence of moxibustion on the activation of NLRP3 inflammasome and its mechanism in treating ulcerative colitis by observing miR7/RNF183 inducing IκB α ubiquitination to regulate NF-κB signaling pathway in an ulcerative colitis rat model.
Methods:
An ulcerative colitis rat model was established by unlimited access to self-administration of 3.5% (w/v) dextran sulfate sodium solution. Mild moxibustion was applied to bilateral Tianshu points (ST25) in the moxa-stick moxibustion group; rats in the control group were intervened by intraperitoneal injection of ubiquitination inhibitor, MG132. The disease activity index was determined at the end of the intervention; colon injury was observed and scored after hematoxylin-eosin staining; the immunohistochemical method was adopted to detect the expressions of colonic IL-1β and NLRP3 proteins; Western blot determined the expressions of RNF183, IκB α, and NF-κB p65 proteins in the colon; the immunofluorescence test was used to observe the coexpression of IκB α/ubiquitin and IκB α/RNF183 proteins in the colon; immunoprecipitation assay was adopted to observe the interaction between IκB α and RNF183 proteins; and quantitative real-time polymerase chain reaction determined the expression of colonic miR7.
Results:
Moxibustion lowered the disease activity index, manifesting as restored colonic tissue and reduced inflammatory reaction, and decreased expression levels of NLRP3 and IL-1β proteins, compared with the model group. It also reduced colonic expression of NF-κB p65 protein, together with the increased level of IκB α protein and weaker expression levels of ubiquitin and RNF183 proteins and mRNAs and stronger expression of miR7. There were no significant differences between the moxa-stick moxibustion group and the control group except the expressions of RNF183 protein and mRNA and miR7.
Conclusion:
Moxibustion encourages the recovery of colon injury probably by regulating the expression of NLRP3 protein in ulcerative colitis rats through miR7/RNF183/NF-κB signaling pathway.
Insights
Moxibustion treatment effectively reduces ulcerative colitis symptoms in rats by modulating the miR7/RNF183/NF-κB pathway. This approach lowers NLRP3 inflammasome activation and promotes colon tissue recovery.
Area of Science:
- Gastroenterology
- Immunology
- Traditional Chinese Medicine
Background:
- Ulcerative colitis (UC) is a chronic inflammatory bowel disease with unclear mechanisms.
- Moxibustion is a recognized therapy for UC, but its underlying molecular pathways require elucidation.
Purpose of the Study:
- To investigate the mechanism of moxibustion in treating ulcerative colitis (UC) in a rat model.
- To explore the role of miR7/RNF183 in inducing IκBα ubiquitination and regulating the NF-κB signaling pathway.
Main Methods:
- Established a UC rat model using dextran sulfate sodium (DSS).
- Applied mild moxibustion to Tianshu points (ST25) and used MG132 as a control.
- Assessed disease activity index, colon injury scores, and protein/mRNA expression (NLRP3, IL-1β, RNF183, IκBα, NF-κB p65, ubiquitin) via immunohistochemistry, Western blot, immunofluorescence, and qPCR.
- Investigated protein interactions using immunoprecipitation assays.
Main Results:
- Moxibustion significantly reduced disease activity index and colon injury, decreasing NLRP3 and IL-1β expression.
- Moxibustion lowered NF-κB p65, increased IκBα, and decreased ubiquitin and RNF183 expression, while upregulating miR7.
- The miR7/RNF183/NF-κB pathway was identified as a key target of moxibustion therapy.
Conclusions:
- Moxibustion promotes colon injury recovery in ulcerative colitis rats.
- The therapeutic effect is likely mediated by regulating NLRP3 inflammasome activation via the miR7/RNF183/NF-κB signaling pathway.
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