Herbs-partitioned moxibustion improves intestinal epithelial tight junctions by upregulating A20 expression in a mouse model of Crohn's disease

Yin Shi; Yajing Guo; Jing Zhou; Luyi Wu; Liu Chen; Yi Sun; Tao Li; Jimeng Zhao; Chunhui Bao; Huangan Wu

doi:10.1016/j.biopha.2019.109149

Herbs-partitioned moxibustion improves intestinal epithelial tight junctions by upregulating A20 expression in a mouse model of Crohn's disease

Biomed Pharmacother. 2019 Oct:118:109149. doi: 10.1016/j.biopha.2019.109149. Epub 2019 Jul 12.

Authors

Yin Shi¹, Yajing Guo², Jing Zhou³, Luyi Wu⁴, Liu Chen⁵, Yi Sun⁶, Tao Li⁷, Jimeng Zhao⁸, Chunhui Bao⁹, Huangan Wu¹⁰

Affiliations

¹ Key Laboratory of Acupuncture and Immunological Effects, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China; Outpatient Department, Shanghai Research Institute of Acupuncture and Meridian, Shanghai, 200030, China. Electronic address: flysy0636@163.com.
² Graduate School, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: guoyajing0817@163.com.
³ Graduate School, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: 419045934@qq.com.
⁴ Key Laboratory of Acupuncture and Immunological Effects, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China; Qigong Institute, Shanghai University of Traditional Chinese Medicine, Shanghai, 200030, China. Electronic address: luyitcm@163.com.
⁵ Graduate School, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: willow_chen0913@163.com.
⁶ Graduate School, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: 1025830709@qq.com.
⁷ Graduate School, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China. Electronic address: 1277713209@qq.com.
⁸ Key Laboratory of Acupuncture and Immunological Effects, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China; Key Laboratory of Acupuncture and Immunological Effects, Shanghai Research Institute of Acupuncture and Meridian, Shanghai, 200030, China. Electronic address: zhaojm0414@163.com.
⁹ Key Laboratory of Acupuncture and Immunological Effects, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China; Key Laboratory of Acupuncture and Immunological Effects, Shanghai Research Institute of Acupuncture and Meridian, Shanghai, 200030, China. Electronic address: baochunhui789@126.com.
¹⁰ Key Laboratory of Acupuncture and Immunological Effects, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China; Key Laboratory of Acupuncture and Immunological Effects, Shanghai Research Institute of Acupuncture and Meridian, Shanghai, 200030, China. Electronic address: wuhuangan@126.com.

PMID: 31302421
DOI: 10.1016/j.biopha.2019.109149

Abstract

Background: To investigate effects moxibustion exerts on A20 expression and regulation of intestinal epithelial tight junctions via the TNF-α-NF-κB-MLCK pathway in Crohn's disease (CD).

Methods: C57BL/6 wild type (WT) and A20^IEC-KO mice (48 each) were randomly divided into normal control (NC), model control (MC), mesalazine (MESA) and herbs-partitioned moxibustion (HPM) groups (12 mice per group). An experimental model of CD was established using 2, 4, 6 trinitrobenzene sulfonic acid. MESA and HPM mice were treated with MESA and HPM (at Tianshu (ST25) and Qihai (CV6)), respectively. In HPM group, moxa cones (0.5 cm in diameter and 0.3 cm in height) made of refined mugwort floss were placed on herbal cakes (medicinal formula dispensing [radix] Aconiti praeparata, [cortex] Cinnamomi, etc.) at Tianshu (ST25) and Qihai (CV6) and ignited. The moxa cones were ignited, and two moxa cones were used for each treatment once daily for 10 days. In MESA group, mice were fed MESA, which was prepared at a proportion of 1:0.0026, twice daily for 10 days.

Results: Intestinal epithelial ultrastructure of WT HPM mice improved more than A20^IEC-KO HPM mice compared to MC mice. WT HPM mice exhibited greater expression of A20 compared with MC mice (P < 0.01). TNF-α, NF-kB p65, MLCK, MLC, TRAF6 and RIP1 levels in A20^IEC-KO and WT HPM mice were all decreased compared to MC mice (P_all < 0.01). NF-κB p65、MLCK and TRAF6 levels were increased in A20^IEC-KO HPM mice as compared to WT HPM mice (P_all < 0.05). Intestinal epithelial levels of occludin, claudin-1, ZO-1 and F-actin increased in all HPM mice (P_all < 0.01-0.05), while occludin, claudin-1, and ZO-1 levels were lower in A20 ^IEC-KO HPM mice (P < 0.05, P < 0.01, P < 0.01).

Conclusion: HPM downregulates abnormal activation of the TNF-α-NF-κB-MLCK pathway by upregulating expression of A20 in a mouse model of CD, thereby protecting intestinal epithelial tight junctions and repairing the damage CD causes to the intestinal epithelial barrier.

Keywords: A20; Crohn’s disease; Herbs-partitioned moxibustion; Intestinal epithelial barrier; TNF-α-NF-κB-MLCK pathway.

MeSH terms

Animals
Artemisia / chemistry*
Colon / metabolism
Colon / ultrastructure
Crohn Disease / metabolism
Crohn Disease / pathology
Crohn Disease / therapy*
Disease Models, Animal
Epithelial Cells / metabolism
Epithelial Cells / pathology
Intestinal Mucosa / metabolism
Intestinal Mucosa / ultrastructure*
Mice, Inbred C57BL
Mice, Knockout
Moxibustion / methods*
Permeability
Tight Junctions / metabolism
Tight Junctions / ultrastructure*
Tumor Necrosis Factor alpha-Induced Protein 3 / genetics
Tumor Necrosis Factor alpha-Induced Protein 3 / metabolism*
Up-Regulation

Substances

Tumor Necrosis Factor alpha-Induced Protein 3
Tnfaip3 protein, mouse