Moxibustion attenuates inflammation and alleviates axial spondyloarthritis in mice: Possible role of APOE in the inhibition of the Wnt pathway

Xiao Xu; Huan Yang; Jun-Ning Chen; Li-Jiang-Shan Hua; Rong-Yun Wang; Ting Liu; Ya-Nan Shi; Qing-Feng Wu; Xi Liu; Hong-Yuan Wang; Zhi-Ling Sun; Hong Zhang; Qiu-Hua Sun

doi:10.1016/j.jtcme.2022.04.002

Moxibustion attenuates inflammation and alleviates axial spondyloarthritis in mice: Possible role of APOE in the inhibition of the Wnt pathway

J Tradit Complement Med. 2022 Apr 22;12(5):518-528. doi: 10.1016/j.jtcme.2022.04.002. eCollection 2022 Sep.

Authors

Xiao Xu¹, Huan Yang², Jun-Ning Chen¹, Li-Jiang-Shan Hua¹, Rong-Yun Wang¹, Ting Liu¹, Ya-Nan Shi¹, Qing-Feng Wu³, Xi Liu⁴, Hong-Yuan Wang⁵, Zhi-Ling Sun⁶, Hong Zhang⁷, Qiu-Hua Sun¹

Affiliations

¹ School of Nursing, Zhejiang Chinese Medical University, Hangzhou, Zhejiang Province, 310000, China.
² First Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou, Zhejiang Province, 310000, China.
³ Third Clinical Medical College, College of Acupuncture and Moxibustion, Zhejiang Chinese Medical University, Hangzhou, Zhejiang Province, 310000, China.
⁴ College of Basic Medicine, Zhejiang Chinese Medical University, Hangzhou, Zhejiang Province, 310000, China.
⁵ Department of Rheumatology, Affiliated hospital of Nanjing university of Chinese Medicine, Nanjing, Jiangsu Province, 210000, China.
⁶ School of Nursing, Nanjing university of Chinese Medicine, Nanjing, Jiangsu Province, 210000, China.
⁷ Department of Nursing, The Affiliated Changzhou No.2 People's Hospital of Nanjing Medical University, Changzhou, Jiangsu Province, 213000, China.

Abstract

Background and aim: Moxibustion is widely used in China and other East Asian countries to manage the symptom of ankylosing spondylitis (AS). This study investigated the effects of moxibustion intervention on protein expression through proteomics analysis in AS mice.

Experimental procedure: Proteoglycan-induced spondylitis (PGISp) was established in Balb/c mice. PGISp mice were intervened with daily moxibustion at ST36, BL23, and DU4 for four weeks. Various biochemical (including pro-inflammatory cytokines and bone metabolism indexes) and histopathological parameters were determined. The effects of moxibustion on protein changes in AS mice were analyzed using data-independent acquisition-mass spectrometry (DIA-MS). The target proteins were then confirmed by Western blot analysis.

Results: Moxibustion significantly decreased pro-inflammatory cytokine expression including IL-1β, TNF-α, IL-17, and IL-6, reduced the mRNA expression of RANKL, RANK, ALP, and OCN, and improved the histopathological examination in AS mice. DIA-MS proteomic technique has identified 25 candidate proteins involved in the mechanisms of moxibustion for AS mice, most of which are mainly associated with the regulation of Wnt/β-catenin. Integrated pathway analysis revealed that glycine, serine and threonine metabolism together with lipid metabolism were the most important canonical pathways involved in the anti-AS effect of moxibustion. In line with the multi-omic data, the levels of BPGM, APOC₂, APOE, and GPD1 modified in the AS mice, intervened with moxibustion as confirmed by Western blot. In particular, APOE may play a key role in linking the lipid metabolism and the Wnt/β-catenin pathway of new bone formation.

Conclusion: In conclusion, moxibustion may reduce pro-inflammatory cytokines and improve bone erosion for AS mice. The regulation of APOE by moxibustion may have a potential inhibitory effect on the Wnt/β-catenin pathway in AS mice. However, due to the lack of silencing or overexpression of key molecules of the signal pathway, whether the beneficial and positive effect of moxibustion involved in the regulation of Wnt/β-catenin signaling pathway by APOE or other aspects, needed to be explored in further study.

Keywords: Data-independent acquisition; Moxibustion; Proteoglycan-induced spondylitis; Wnt/β-catenin signaling pathway.