[Frontiers in Bioscience, Landmark, 25, 498-512, Jan 1, 2020]

T-96 attenuates inflammation by inhibiting NF-κB in adjuvant-induced arthritis

Yun Yu1, Bian Yong2, Changliang Xu2, Luyong Zhang1,3,4,5

1Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing 210009, China, 2Laboratory Animal Center of Nanjing University of Chinese Medicine, Nanjing 210023, China, 3Jiangsu Center for Pharmacodynamics Research and Evaluation, China Pharmaceutical University, Nanjing 210009, China, 4Key Laboratory of Drug Quality Control and Pharmacovigilance, China Pharmaceutical University, Ministry of Education, Nanjing 210009, China, 5Center for Drug Screening and Pharmacodynamics Evaluation, School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
    3.1. Chemicals and reagents
    3.2. Isolation and culture of fibroblast-like synoviocytes
    3.3. Cell viability assay
    3.4. ELISA
    3.5. Nitric oxide (NO) measurement
    3.6. Western blotting
    3.7. Arthritic rats
    3.8. Experimental procedure
    3.9. Histopathological examination
    3.10. Immunohistochemical assessment
    3.11. Statistical analysis
4. Results
    4.1. Evaluation of cytotoxicity in normal FLSs
    4.2. T-96 attenuates IL-1β-induced inflammatory signaling
    4.3. T-96 attenuates IL-1β-induced target protein expression and NO production
    4.4. T-96 decreases paw swelling, arthritis indices, and weight loss in arthritic rats
    4.5. Effects of T-96 on histopathological changes in arthritic rats
    4.6. Effects of T-96 on the expression of IL-10 and IL-17 in joint synovium in arthritic rats
    4.7. Effects of T-96 on serum levels of cytokines in arthritic rats
5. Discussion
6. Acknowledgments
7. References

1. ABSTRACT

The extract of the medicinal plant, Tripterygium wilfordii Hook. f. (TW), has been used in the treatment of diverse autoimmune diseases, including rheumatoid arthritis. However, the high frequency of toxic side effects has limited its clinical use. In order to reduce toxicity without losing the therapeutic benefit, the pharmacological activity and toxicity of four compounds (T-96, triptolide, neotripterifordin, and tripterifordin) from TW were evaluated. The current study revealed that these compounds interfere with the IL-1β signaling pathway, which stimulates the secretion of pro-inflammatory cytokines (IL-6) in primary rheumatoid arthritis synovial fibroblasts (RASFs). These compounds inhibit IL-6 production, and among these, T-96 was the most effective. Moreover, T-96 blocks activation of NF-kappa B and p38 and ameliorates the joint destruction and the clinical signs of the disease in adjuvant-induced arthritic rats. These data suggest that among the four compounds of the TW, T-96 possesses highest anti-rheumatoid arthritis activity though inhibiting IL-1-mediated inflammatory signaling pathways.

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Key Words: T-96, NF-kappaB, Rheumatoid Arthritis, Fibroblast-like synoviocytes

Send correspondence to: Luyong Zhang, Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing 210009, China. E-mail: lyzhang@cpu.edu.cn; Changliang Xu, PhD. Laboratory Animal Center of Nanjing University of Chinese Medicine, Nanjing 210023, China, Tel: 025-85811256, Fax: 025-83271142, E-mail: cl.xu81@gmail.com