Changes of microenvironment in a rat model of arthritis treated by the combination of methotrexate and diclofenac sodium

doi:10.3969/j.issn.2095-4344.0368

Abstract

Abstract:

BACKGROUND: Rheumatoid arthritis is an autoimmune disorder that typically results in swollen and painful facet joints and morning stiffness. Its drug therapy is mainly the combination of non-steroidal anti-inflammatory drugs and anti-rheumatic drugs, however, the drug effects on the T-lymphocyte subset balance and the underlying mechanism remain unclear.
OBJECTIVE: To explore the effect of the combination of methotrexate and diclofenac sodium on the balance of Th17/Treg cells in a rat model of collagen-induced arthritis.
METHODS: After the male Wistar rat model of collagen-induced arthritis model was established, they were randomly divided into blank control, model, methotrexate, diclofenac sodium, methotrexate + diclofenac sodium, and positive control groups. They were assessed by measuring the arthritis index and toe swelling degree. After 4-week treatment, the expression levels of interleukin 10 and transforming growth factor β in the peripheral blood was detected by ELISA, and the change in Th17/Treg cells in the spleen was tested by flow cytometry; the mRNA expression levels of Foxp3 and ROR-γt in the spleen were examined by RT-PCR.
RESULTS AND CONCLUSION: Compared with the model group, methotrexate, diclofenac sodium, and their combination could improve the general condition, reduce toes swelling degree, lower arthritis index (especially the methotrexate + diclofenac sodium group), reduce inflammation cell infiltration, and inhibit ROR-γt regulating Th17 cell differentiation and regulating the Foxp3-induced high differentiation of CD4+CD25+Foxp3+Tre, thereby restoring Treg/Th17 cell balance and improving the arthritis symptoms. The average gray value was significantly different among groups (P < 0.05). Except the down-regulation of interleukin 10 and interleukin 6 levels like diclofenac sodium, the combined therapy could down-regulate the expression level of transforming growth factor β, increase the Treg/Th17 cell proportion, and inhibit the expression of Foxp3 and ROR-γt. Additionally, methotrexate significantly reduced the expression level of transforming growth factor β. In summary, the combination of methotrexate and diclofenac sodium possesses a synergistic effect, which provides the theoretical foundation for the treatment of rheumatoid arthritis.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Arthritis, Methotrexate, Th17 Cells, T-Lymphocytes, Cytokines, Tissue Engineering

CLC Number:

R318

Niu Zi-han1, Ran Xin-jian2, Mawlan•Mamut1, Pan Chuan-peng1, He Dong-hang3, Ren Cong-gai3, Xu Qi4. Changes of microenvironment in a rat model of arthritis treated by the combination of methotrexate and diclofenac sodium[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(32): 5163-5168.

Figures/Tables 3

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