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Chinese Herbal Formula Ermiao Powder (二妙散) Regulates Cholinergic Anti-inflammatory Pathway in Rats with Rheumatoid Arthritis

Abstract

Objective

To investigate the effect of Chinese herbal formula Ermiao Powder (二妙散, EMP) on the expression of cholinergic anti-inflammatory pathway in rats with rheumatoid arthritis (RA).

Methods

Seventy-two rats were randomly divided into 6 groups according to body weight, including normal control group, collageninduced arthritis (CIA) group, three doses EMP groups, and methotrexate (MTX) group (n=12 per group). All of the rats except for those in the normal control group were given multipoint subcutaneous injection of bovine type II collagen to establish a CIA model. Three EMP groups received a high- (4.5 g/kg), medium- (3.0 g/kg), and low- (1.5 g/kg) doses of EMP by intragavage, respectively. MTX group was injected intraperitoneally MTX at 0.9 mg/kg once a week as the positive control. The administration was 3 consecutive weeks. Joint swelling, arthritis index, and body weight changes in different experimental groups of rats were tested. The joint damage was evaluated by masson staining. Quantitative real-time polymerase chain reaction, Western blot, and immunohistochemistry (IHC) were performed to evaluate the expression of CHRNA7, encoding α7 nicotinic acetylcholine receptor in the cholinergic anti-inflammatory pathway, in different tissues and their localization in the spleen and joints.

Results

CHRNA7 expression levels were significantly higher in the joints and spleens of CIA group than those in normal control group (both P<0.05). Moreover, the CHRNA7 mRNA and protein levels in the spleen and joints of MTX and three doses of EMP groups were significantly lower than CIA group (all P<0.05). Compared with the MTX group, treatment with low-dose EMP resulted in significant reduction of CHRNA7 mRNA and protein expression levels (P<0.05 or P<0.01). IHC showed positive signals of CHRNA7 in the white pulp and red pulp of the spleens of rats; CHRNA7 was expressed on fibroblast-like synoviocytes, macrophages, and endothelial cells in the joints of rats, and the expression in the joints of low-dose EMP group was significantly lower than that in the CIA group (P<0.01).

Conclusions

Cholinergic anti-inflammatory pathway was involved in the generation of the inflammatory reaction in CIA rats, and EMP exerted therapeutic effect on RA through cholinergic anti-inflammatory pathway.

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Authors and Affiliations

Authors

Contributions

Hao HQ designed the experiment. Li Z conducted the experiments and wrote the paper. Gao YT analysed the data. Liu J and Wang Z were responsible for reproduction and breeding of the animal model.

Corresponding author

Correspondence to Hui-qin Hao.

Additional information

Conflict of Interest

The authors declare that there are no conflicts of interest.

Supported by the National Natural Science Foundation of China (No. 81904034), the Basic Research Project of Shanxi Province (No. 201801D121228), the Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi (No. 201804001), and the Research Project of Health Commission of Shanxi Province (No. 2018001)

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Li, Z., Liu, J., Hao, Hq. et al. Chinese Herbal Formula Ermiao Powder (二妙散) Regulates Cholinergic Anti-inflammatory Pathway in Rats with Rheumatoid Arthritis. Chin. J. Integr. Med. 26, 905–912 (2020). https://doi.org/10.1007/s11655-020-3471-2

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  • DOI: https://doi.org/10.1007/s11655-020-3471-2

Keywords

  • Ermiao Powder
  • rheumatoid arthritis
  • α7nAchR
  • CHRNA7
  • Chinese medicine